NZ617736B2 - Processing biomass - Google Patents

Abstract

Disclosed is a method for producing a sugar solution from biomass comprising: treating the biomass to reduce its recalcitrance, thereby making treated biomass, wet milling and simultaneously jet mixing the treated biomass to make wet milled biomass; saccharifying the wet milled biomass, thereby producing the sugar solution. Also disclosed is a wet milling system comprising one or more jet heads and one or more wet milling heads in a tank comprising fluid having biomass dispersed therein. ucing the sugar solution. Also disclosed is a wet milling system comprising one or more jet heads and one or more wet milling heads in a tank comprising fluid having biomass dispersed therein.

Description

PROCESSING BIOMASS RELATED APPLICATIONS This application claims priority to US. ional Application Serial No. 61/495,217, filed June 9, 201 1. The complete disclosure of this provisional application is hereby incorporated by reference herein.

BACKGROUND Cellulosic and lignocellulosic materials are produced, processed, and used in large ‘10 quantities in a number of applications. Often such materials are used once, and then discarded as waste, or are simply considered to be waste materials, e.g., sewage, e, sawdust, and stover.

SUMMARY Processes are sed herein for saccharifying or ying a biomass material, e.g., cellulosic, lignocellulosic and/or starchy feedstocks, by converting s material to low molecular weight sugars, e.g., saccharifying the ock using an enzyme, e.g., one or more cellulase and/or amylase. The invention also relates to ting a feedstock to a product, e.g., by bioprocessing, such as fermentation. The processes include wet milling a feedstock. The inventors have found that wet milling the feedstock tends to reduce the time required for saccharification, and increase the concentration of sugar that can be obtained in a given saccharification time. Wet milling alone or working synergistically with any treatment described herein can reduce recalcitrance of a biomass The processes disclosed herein can utilize low bulk density materials, for example cellulosic or lignocellulosic ocks that have been physically pretreated to have a bulk density ofless than about 0.75 g/cm3, e.g., less than about 0.7, 0.65, 0.60, 0.50, 0.35, 0.25, 0.20, 0.15, 0.10, 0.05. or less, e.g., less than 0.025 g/cm3.

Such materials can be difficult to se in liquids, e.g., with water or a solvent system for saccharification, fermentation, or other processing. Due to their low bulk density, the materials tend to float on the surface of the liquid rather than being wetted out and dispersed into the liquid. In some cases, the materials can be hydrophobic, highly crystalline, or otherwise difficult to wet. At the same time, it is desirable to process the feedstock in a relatively high solids level dispersion, in order to obtain a high final concentration of sugar in the saccharified material, or a high tration of the desired product after processing (e.g., of ethanol or other alcohol(s) after fermentation). In some cases, utilizing the methods described herein the solids level of the dispersion during processing can be, for example, at least 10, 15, 20, 22.5, 25, 27.5, 30, 35, 40, 45, or even at least 50 percent by weight dissolved solids. For example, the solids level can be from about 10 to 50%, e.g., about 10-40%, 10-30%, or 10-20%.

In one , the invention features reducing the particle size of a lignocellulosic al to less than 3000 µm, e.g. less than 2000 µm, less than 1000 µm or even less than 500 µm, e.g., less than 250 µm or less than 100 µm. The particle size range can be between 100 - 3000 µm, e.g., 200–2000 µm, 200–1000 µm, 500- 1000 µm.

In one aspect, the invention features reducing recalcitrance of a lignocellulosic material and wet milling the ellulosic material. In some cases, recalcitrance is reduced prior to wet milling. The material can be densified prior to reducing the recalcitrance or after reducing the itrance and prior to wet milling the material.

In another aspect, the invention features a method comprising wet milling a lignocellulosic material, e.g., a ellulosic al having a reduced itrance.

In another aspect, the invention features a method for producing a sugar solution from biomass comprising: treating the biomass to reduce its recalcitrance, thereby making treated biomass, wet milling and simultaneously jet mixing the treated biomass to make wet milled biomass; saccharifying the wet milled biomass, thereby producing the sugar solution.

Either of these aspects of the invention can include, in some implementations, any of the following features.

The recalcitrance of the biomass material, e.g., a lignocellulosic material, can be reduced, for example, by irradiating the lignocellulosic al, e.g., by ng the material to an on beam. In some cases, ating comprises delivering a dose of at least 5 Mrad to the lignocellulosic material, e.g., at least 10, 20, 30, 50, 100, 150 or even 200 Mrad. For example, doses can be in the range of 5-200 Mrad, e.g., 5- 100 Mrad, 5-50 Mrad, 5-10 Mrad, 10-100 Mrad, or 10-50 Mrad.

The lignocellulosic material may be, for example, a material is selected from the group consisting of wood, particle board, sawdust, agricultural waste, sewage, silage, grasses, rice hulls, e, cotton, jute, hemp, flax, bamboo, sisal, abaca, straw, wheat 2012/041382 straw, corn cobs, corn stover, switchgrass, alfalfa, hay, coconut hair, seaweed, algae, and es thereof.

The biomass may also be combinations of starchy, ellulosic and/or cellulosic materials. For example, a biomass can be an entire plant or part(s) ofa plant e.g., a wheat plant, cotton plant, a corn plant, rice plant or a tree.

In some implementations, wet milling is performed using a rotor/stator head. The rotor and stator may include nesting rings of teeth. In some cases, the stator comprises two or more concentric rings of teeth, and the rotor ses a ring of teeth red to fit n adjacent rings of teeth of the stator. The clearance between the rotor and stator is generally small, to generate high shear, and may be, for example from about 0.01 to 0.25 inches (0.25 to 6.4 mm). The spacing between the teeth in each ring of teeth is also generally small, e.g., from about 0.1 to 0.3 inch (2.5 to 7.6 mm).

Wet milling may be performed using a ity of rotor/stator heads, e.g., when the process is performed in a large tank or vessel.

Wet milling is generally performed at a relatively high shear rate. The shear rate may be, for example, at least 20,000 sec'], (e.g., at least 25,000 sec], at least 30,000 sec"’ at least 40,000 sec'1 or at least 50,000 sec'l). The shear rate can be, for example from about 30,000 sec’1 to about 50,000 sec'l (e.g., from about 25,000 sec'1 to about 50,000 sec'l, from about 30,000 sec"| to about 50,000 sec'], from about 35,000 sec" to about 50,000 sec", from about 40,000 sec'1 to about 50,000 sec'l, from about 20,000 sec'1 to about 45,000 see", from about 20,000 sec'1 to about 40,000 see", from about 20,000 sec‘ to about 30,000 sec", from about 30,000 sec" to about 40,000 sec").

In some implementations, wet milling is performed in-line. A jet mixer may be applied during wet milling. The jet mixer may also be used during subsequent processing, e. g., during fermentation. The method may further include adding an enzyme to the biomass material, e.g., a lignocellulosic material, before, during or after wet milling, and/or adding a microorganism to the biomass material or a sugar derived from the biomass material. In some cases, the microorganism is added after wet milling has been completed, e.g., to avoid damage to the microorganism from wet milling. In some implementations, the rganism converts the biomass feedstock or sugar to a product 2012/041382 selected from the group consisting of alcohols, organic acids, sugars, hydrocarbons, and mixtures thereof.

The methods described herein generally provide relatively rapid and effective sing of a relatively high solids level of feedstock. By increasing the initial solids level of feedstock in the mixture, the process can proceed more y, more efficiently and more cost—effectively, and a high resulting concentration can generally be achieved in the final product. In some cases, solids may be removed during saccharification, e.g., by a centrifuge, and more feedstock may be added. The removed solids may be used as a t, e.g., as a combustible fuel for cogeneration of electricity and/or as an animal 1O feed.

The saccharification processes described herein allow biomass material, e.g., a cellulosic or lignocellulosic feedstock, to be converted to a convenient and concentrated form which can be easily orted and utilized in another manufacturing facility, e.g., a facility configured to ferment sugar solutions to alcohols, to cture a product, e. g., a fuel such as ethanol, butanol or a hydrocarbon. Such trates can use less water, which can result in ntial manufacturing and transportation cost savings.

Some ses disclosed herein include saccharification of the feedstock, and transportation of the feedstock from a remote location, e. g., where the feedstock is produced or stored, to the manufacturing facility. In some 'cases, saccharification can take place partially or entirely during transport.

In some cases, the systems described herein, or ents thereof, may be portable, so that the system can be transported (e.g., by rail, truck, or marine vessel) from one location to another. Such mobile processing is described in US. Serial No. 12/374,549 filed January 21, 2009 and International Application No. WO 11598, the full disclosures of which are incorporated herein by reference.

Exemplary products that can be produced by employing the s described herein include arbons, ns, alcohols (e.g., a monohydric alcohols or a dihydric alcohols), such as ethanol, isobutanol, n—propanol or n-butanol, carboxylic acids, such as acetic acid, lactic acid, citric acid, propionic acid, succinic acid, 3—hydroxyproprionic acid or butyric acid, salts ofa carboxylic acid, a mixture of carboxylic acids and salts of carboxylic acids and esters of carboxylic acids (e.g., methyl, ethyl and n—propyl esters), ketones, aldehydes, alpha unsaturated acids, beta unsaturated acids, such as acrylic acid, olefins, such as ethylene, butenes, and mixtures of any of these. Specific examples include ethanol, propanol, propylene glycol, butanol, 1,4-butanediol, 1,3- propanediol, methyl or ethyl esters of any of these alcohols, methyl te, methylmethacrylate, ts also include sugars, e.g., glucose, xylose and xylitol.

These and other products are described in U.S. Serial No. ,900 filed April 3, 2009; the full disclosure of which is incorporated by reference herein.

In one aspect, the invention features a wet milling system comprising a wet mill disposed in a fluid having a biomass material dispersed n. The system can, for example, be used for processing lingocellulosic material that has optionally been irradiated (e.g., with an electron beam). The system can e a jet mixer disposed in the fluid. The wet milling systems can include a rotor/stator head, for example with the rotor and stator including nesting rings of teeth. Furthermore, the stator can have two or more concentric rings of teeth. Other aspects of the invention include a tank with one or more jet head, and one or more wet mill disposed in the tank.

In another aspect, the invention features a wet g system comprising one or more jet heads and one or more wet milling heads in a tank comprising fluid having s dispersed n.

Bulk density is determined using ASTM D1895B. Briefly, the method involves filling a measuring cylinder of known volume with a sample and obtaining a weight of the sample. The bulk density is ated by dividing the weight of the sample in grams by the known volume of the cylinder in cubic centimeters.

All publications, patent applications, patents, and other references mentioned herein or attached hereto are incorporated by reference in their entirety for all that they contain.

DESCRIPTION OF THE DRAWINGS is a diagram illustrating the enzymatic hydrolysis of cellulose to glucose. is a flow diagram illustrating conversion of a feedstock to sugars and other products. is a diagrammatic ration of a saccharification system according to one embodiment. is a diagrammatic ration of a saccharification system ing to another embodiment. is a schematic diagram of an ethanol manufacturing facility that has been retrofitted to utilize the solutions and suspensions disclosed herein.

WO 70707 is a top plan view of the led rotor and stator ofa wet milling head according to one embodiment. is an enlarged section view of showing the clearance between the rotor and stator. is a perspective view of the rotor and stator together. is an exploded perspective of the rotor and stator. is a bottom plan view of the rotor taken along view-line 7-7 of is a top plan View of the stator taken along view line 8-8 of is an ed view of the area of the rotor indicated in is an enlarged view of the area of the stator indicated in 1O 1 is a top plan view of the assembled rotor and stator ofa wet milling head according to a second embodiment. A is an enlarged section view of g the clearance between the rotor and stator. is a perspective view of the rotor and stator together. is an exploded perspective of the rotor and stator. is a bottom plan view of the rotor taken along view line 14—14 of . is a top plan view of the stator taken along view line 15-15 of . is an enlarged view of the area ofthe rotor indicated in . is an enlarged view of the area of the stator indicated in .

FIGS. 18 and 18A are diagrams illustratingjet flow exiting a jet mixer nozzle. is a diagrammatic ctive view of a jet-flow agitator according to one embodiment. A is an enlarged perspective View of the impeller and jet tube of the jet—flow agitator of . B is an enlarged perspective View of an ate impeller. is a cross-sectional view ofa system for wet milling.

DETAILED DESCRIPTION Using the methods described herein, biomass (e.g., plant biomass, animal biomass, paper, and municipal waste biomass) can be sed to produce useful ediates and products such as organic acids, salts of organic acids, anhydrides, esters of organic acids and fuels, e.g., fuels for internal combustion engines or feedstocks for fuel cells. Systems and processes are described herein that can use as feedstock cellulosic and/or lignocellulosic materials that are readily ble, but often can be difficult to process, e.g., municipal waste streams and waste paper streams, such as streams that include newspaper,- kraft paper, corrugated paper or mixtures of these.‘ Generally, if required, als can be physically treated for processing and/or after processing, often by size reduction. Many ofthe processes described herein can effectively lower the recalcitrance level of the feedstock, making it easier to process, such as by bioprocessing (e.g., with any microorganism described herein, such as a homoacetogen or a heteroacetogen, and/or any enzyme described herein), thermal processing (e.g., gasification or pyrolysis) or chemical s (e.g., acid hydrolysis or oxidation). Biomass feedstock can be treated or processed using one or more of any of the methods described herein, such as mechanical ent, chemical ent, radiation, sonication, oxidation, pyrolysis or steam explosion. The various treatment systems and methods can be used in combinations of two, three, or even four or more of these technologies or others described herein and elsewhere. In some instances wet milling alone can reduce itrance or act synergistically or with other treatment processes described herein.

The processes disclosed herein can utilize low bulk density materials, for e osic or lignocellulosic feedstocks that have been physically ated to have a bulk density of less than about 0.75 g/cm3, e.g., less than about 0.7, 0.65, 0.60, 0.50, 0.35, 0.25, 0.20, 0.15, 0.10, 0.05. or less, e.g., less than 0.025 g/cm3. Bulk density is determined using ASTM . Briefly, the method involves filling a measuring cylinder of known volume with a sample and obtaining a weight of the . The bulk density is calculated by dividing the weight of the sample in grams by the known volume of the cylinder in cubic centimeters. If desired, low bulk density materials can be densified, for example, by s described in US. Patent 809 the full disclosure of which is hereby incorporated by reference.

Saccharification In order to convert the feedstock to a form that can be readily processed, the glucan- or xylan-containing cellulose in the feedstock is hydrolyzed to low molecular weight carbohydrates, such as sugars, by a saccharifying agent, e.g., an enzyme or acid, a process referred to as saccharification. The low lar weight carbohydrates can then be used, for example, in an existing manufacturing plant, such as a single cell protein plant, an enzyme manufacturing plant, or a fuel plant, e.g., an ethanol manufacturing facility.

The feedstock is combined with the saccharifying agent in a liquid medium, e.g., a solvent such as an aqueous on, and the mixture is wet milled. Methods for wet milling the material in the liquid medium are discussed in detail below. In some implementations, during and/or after wet milling the saccharifying agent, material and liquid medium are mixed using ajet mixer. In some cases jet mixing ues throughout saccharification.

In some implementations, the material and/or the saccharifying agent are added incrementally rather than all at once. For example, a portion of the material can be added to the liquid medium, dispersed therein, and mixed with the rifying agent until the material is at least partially saccharified, at which point a second portion of the material is dispersed in the medium and added to the e. This process can continue until a desired sugar concentration is obtained.

The feedstock can be hydrolyzed using an enzyme, such as a cellulase or an amylase or mixtures of these s. For example, the biomass material can be combined with the enzyme in a solvent, e.g., in an aqueous solution.

Enzymes and biomass-destroying organisms that break down biomass, such as the cellulose and/or the lignin portions of the biomass, contain or manufacture various cellulolytic enzymes (cellulases), ligninases or various small molecule biomass- destroying metabolites. These enzymes may be a complex of enzymes that act synergistically to e crystalline cellulose or the lignin portions of biomass.

Examples of cellulolytic enzymes include: ucanases, cellobiohydrolases, and cellobiases (B-glucosidases). Referring to a cellulosic substrate is initially hydrolyzed by endoglucanases at random locations producing oligomeric intermediates.

These intermediates are then substrates for exo-splitting glucanases such as cellobiohydrolase to produce cellobiose from the ends of the cellulose r.

Cellobiose is a soluble 1,4-linked dimer of glucose. Finally cellobiase cleaves iose to yield e. Suitable cellulases will be sed herein in a later section.

The time required for complete saccharification will depend on the s conditions and the feedstock and enzyme used. If saccharification is performed in a manufacturing plant under controlled conditions, the cellulose may be substantially entirely converted to glucose in about 12—96 hours, e.g., less than 48 hour, less than 36 hours, less than 24 hours, less than 18 hours, less than 12 hours or even less than 8 hours.

If saccharification is med partially or completely in transit, saccharification may take longer.

In some cases, saccharification is performed at a pH of about 4 to 7, e.g., about 4.5 to 6, or about 5 to 6.

It is lly preferred that the final concentration of glucose in the sugar solution be relatively high, e.g., greater than 10 wt.%, or greater than 15, 20, 30, 40, 50, 60, 70, 80, 90 or even greater than 95% by weight. This reduces the volume to be shipped, and also inhibits microbial growth in the solution. After saccharification, the volume of water can be d, e.g., by evaporation or distillation.

A relatively high concentration solution can be obtained by limiting the amount of medium, e.g., water, added to the feedstock with the enzyme. The concentration can be controlled, e. g., by lling how much saccharification takes place. For example, concentration can be sed by adding more feedstock to the solution. In some cases, solids are removed during saccharification, e.g., by fuge, allowing more feedstock to be added. Solubility of the feedstock in the medium can be increased, for e, by increasing the temperature of the solution, and/or by adding a surfactant as will be discussed below. For example, the solution can be maintained at a temperature of 40- 50°C, 50-60°C, 60—80°C, or even higher.

Fermentation Microorganisms can e a number of useful intermediates and ts by fermenting a low molecular weight sugar produced by rifying the treated feedstock. For example, fermentation or other bioprocesses can produce alcohols (e.g., n-butanol, isobutanol, ethanol or erythritol), organic acids (e.g., acetic, butyric, citric or lactic acid), hydrocarbons, hydrogen, proteins or mixtures of any of these materials.

Yeast and Zymomonas bacteria, for example, can be used for fermentation or conversion. Other microorganisms are discussed in the Materials n, below. The optimum pH for fermentations is about pH 4 to 7. The optimum pH for yeast is from about pH 4 to 5, while the optimum pH for Zymomonas is from about pH 5 to 6. Typical fermentation times are about 24 to 168 (e.g., 24-96 hrs) hours with temperatures in the range of 20 °C to 40 °C (e.g., 26 °C to 40 °C), however thermophilic microorganisms prefer higher temperatures.

In some embodiments e. g., when anaerobic organisms are used, at least a portion of the fermentation is conducted in the e of oxygen e.g., under a blanket of an inert gas such as N2, Ar, He, C02 or mixtures f. Additionally, the mixture may have a constant purge of an inert gas flowing through the tank during part of or all of the fermentation. In some cases, anaerobic conditions can be achieved or maintained by carbon dioxide production during the tation and no additional inert gas is needed.

In some embodiments, all or a portion of the fermentation process can be interrupted before the low molecular weight sugar is completely converted to a product (e.g. ethanol). The intermediate tation products include high concentrations of sugar and carbohydrates. The sugars and carbohydrates can be isolated as discussed below. These intermediate fermentation products can be used in preparation of food for human or animal consumption. Additionally or alternatively, the ediate fermentation products can be ground to a fine particle size in a stainless-steel laboratory mill to produce a flour-like substance.

The fermentations include the methods and products that are disclosed in US.

Provisional Application Serial No. 61/579,559, filed December 22, 2011 and US. ional Application Serial No. 61/579,576, filed er 22, 2011 the disclosure of which is orated herein by reference.

Mobile fermenters can be utilized, as described in US. Provisional Patent Application Serial No. 60/832,735, now hed International ation No. WO 2008/011598. Similarly, the saccharification equipment can be mobile. Further, saccharification and/or fermentation may be performed in part or entirely during transit.

Fuel Cells Where the methods described herein produce a sugar solution or suspension, this solution or suspension can subsequently be used in a fuel cell. For example, fuel cells ing sugars derived from cellulosic or ellulosic materials are disclosed in US.

Provisional Application Serial No. 61/579,568, filed er 22, 2011, the complete disclosure of which is incorporated herein by reference.

Thermochemical Conversion Thermochemical conversion can be performed on the d feedstock to produce one or more desired intermediates and/or products. A thermochemical conversion? '10 process includes changing lar structures of carbon-containing material at elevated temperatures. Specific examples include gasification, pyrolysis, reformation, partial oxidation and mixtures of these (in any order).

Gasification converts carbon-containing materials into a synthesis gas (syngas), which can include ol, carbon monoxide, carbon e and en. Many microorganisms, such as acetogens or homoacetogens are capable of utilizing a syngas from the thermochemical sion of s, to produce a product that includes an alcohol, a carboxylic acid, a salt of a carboxylic acid, a carboxylic acid ester or a mixture of any of these. Gasification of biomass (e.g., cellulosic or lignocellulosic materials), can be accomplished by a variety of techniques. For example, gasification can be accomplished utilizing staged steam reformation with a fluidized-bed reactor in which the carbonaceous material is first pyrolyzed in the absence of oxygen and then the pyrolysis vapors are reformed to synthesis gas with steam providing added hydrogen and oxygen.

In such a technique, process heat comes from g char. Another technique utilizes a screw auger reactor in which moisture and oxygen are introduced at the pyrolysis stage and the process heat is generated from g some of the gas produced in the latter stage. Another que utilizes entrained flow reformation in which both external steam and air are introduced in a single-stage gasification reactor. In partial oxidation gasification, pure oxygen is utilized with no steam.

S FOR TREATING A FEEDSTOCK Referring to a process for conversion of a feedstock to sugars and other products, e.g., ethanol, can include, for example, optionally physically pre—treating the feedstock, e.g., to reduce its size (step 110), before and/or after this treatment, ally treating the feedstock to reduce its recalcitrance (step 112), and saccharifying the feedstock to form a sugar solution (step 114). Saccharification can be performed by mixing a dispersion of the feedstock in a liquid medium, e.g., water with an enzyme (step 11 1), as will be discussed in detail below. During or after saccharification, the mixture (if rification is to be partially or completely performed en route) or solution can be transported, e.g., by pipeline, railcar, truck or barge, to a manufacturing plant (step 1 16).

At the plant, the solution can be bio—processed to produce a desired product, e.g., ethanol (step 1 18), which is then processed further, e.g., by distillation (step 120). The individual steps of this process will be bed in detail below. If desired, the steps of measuring lignin content (step 122) and setting or adjusting process ters (step 124) can be performed at various stages of the process, for e just prior to the process step(s) used to change the ure of the feedstock, as shown. If these steps are included, the process parameters are adjusted to compensate for variability in the lignin content of the feedstock, as described in US. Application Series No. 12/704,519 filed in February 1 1, 2010, the complete disclosure of which is incorporated herein by nce.

The mixing step 1 11 and saccharifying step 1 14 can be performed using, for example, either of the s shown in and 2B. These systems include a tank 136, which initially contains a liquid medium and later contains a mixture 138 of liquid medium, feedstock and saccharifying agent. The liquid medium is delivered to the tank through a valved piping system (not shown). The systems also include a hopper 130, in communication with a dispersing unit 134. In the embodiment shown in FIG. ZB, the hopper 130 receives ock that has been treated to reduce its size and optionally to reduce its recalcitrance (steps 110 and 1 12 above) by a feedstock pretreatment module 132. In both ments, the hopper may receive other dry ingredients, such as yeast and nutrients, e.g., from a supply 30. Optionally, a vibrating device 36 may be associated with the hopper, to facilitate delivery of material from the hopper. The system may also optionally include a dispersing unit 134, e.g., ifthe feedstock is difficult to initially wet with the . The liquid medium is drawn into the dispersing unit 134 from the tank, and returned to the tank by the dispersing unit via an outlet pipe 137. The opening of outlet pipe 137 may be above the liquid level, as shown, or may in some instances be submerged in the liquid in the tank. In some cases, depending on the type of milling unit and dispersing unit used, the system may include a pump 139, e.g., a positive displacement pump, configured to ate the liquid medium, and/or a viscometer 141 to monitor the viscosity of the sion and activate the pump when the measured viscosity reaches a predetermined value.

In the embodiment shown in , the feedstock is delivered to the surface of the liquid medium in the tank, e.g., via a delivery device 32 having a delivery conduit 34 (e.g., hose or pipe). The delivery device 32 may also be associated with a vibrating device 36, to facilitate flow of material into the device. The ry device 32 may be, for example, a blower configured to blow fibrous and/or ulate material from a source to a location remote from the source through a hose, e.g., an insulation blower such as the FORCE 3 blower ble from Intec, Frederick, Colorado. Alternatively, the al can be delivered to the surface of the liquid using other techniques, such as gravity feed or a screw conveyor.

In some implementations, the tank is provided with a flexible, air permeable cover, or other device configured to allow air to vent from the tank during delivery of the feedstock, while preventing ock from blowing out of the tank and/or contaminants from entering the tank.

When the particles are generally spherical, e.g., as is the case with hammermilled corn cobs, or otherwise of a morphology that allows them to be easily fed, the feedstock can be gravimetrically fed. For example, the feedstock can be delivered from a hopper above the tank.

As the feedstock material is delivered through delivery conduit 34 onto the surface of the liquid in the tank, liquid is discharged through outlet pipe 137 of the sing unit 134 onto the material. The discharged liquid wets the feedstock material, causing it to sink into the liquid, where it can be sed by the dispersing unit 134 (if one is provided), optionally in combination with the mixing action of a jet mixer 144, discussed below.

Once the feedstock has been delivered to the tank, it is wet milled, using wet milling unit 160, which generally includes a high shear rotor/stator head. Examples of suitable milling units are described in detail below. The wet milling unit can be mounted in any desired location in the tank. It can be side—mounted, as shown, or top and bottom mounted. In some implementations, the wet milling unit can be external to the tank and the tank contents can be pumped through the wet milling unit and ed to the tank. In some cases, the wet milling unit is mounted adjacent to the jet mixing unit 144, described below. In some cases, multiple wet g heads are provided. For e, in a large tank multiple wet milling heads may be mounted at spaced locations within the tank.

Wet milling can be performed in-line or as a batch process.

Wet milling is lly performed at a high shear rate, for example from about ,000 sec'1 to 60,000 sec‘l, or from about 30,000 sec'1 to 50,000 560'].

The wet milling unit may be run for any desired length of time. The wet milling unit can be run in a pulsed manner (e.g., the power to the motor driving the wet g is pulsed), for example the shearing rate can be varied periodically or non-periodically, or, as another example the wet milling unit can be turned on an off repeatedly. Generally, wet milling is discontinued when either the efficiency of saccharification ceases to be improved by wet milling (this can be determined by experimentation for a given set of process parameters), or the shear generated by the wet milling unit causes the temperature of the tank contents to exceed a predetermined m value. The predetermined maximum value may be set, for example, based on the temperature at which the saccharifying agent would be denatured in a short period of time. ng can cause the mean particle size of the biomass material to be reduced.

For example the size can be reduced from about more than 1 mm (e.g. more than 5 mm or more than 10 mm) to less than 1 mm (e.g., less than 0.5mm, less than 0.1 mm or even less than 0.01mm).

In some implementations, the wet milling unit can be used to heat, or partially heat, the tank contents. to a desired processing ature. For e, in one implementation the tank contents are heated by r means to approximately 40°C, 2012/041382 and then the‘wet milling unit is operated for a time ent to raise the temperature to approximately 50°C, a temperature which is advantageous for saccharification. In some cases, wet milling is performed for less than 8 hours, e.g., for I to 4 hours or 1 to 2 hours.

Wet milling may be performed for an even shorter time, e.g., 30 minutes or less. Once this desired temperature is reached the wet milling device is turned off so as to prevent a further increase in temperature. In some cases, the tank contents may be cooled during or after wet milling to prevent overheating. In order to prevent denaturing of the enzymes used in saccharification, it is generally preferred that the tank contents be maintained at or below 50°C, or at least that temperature excursions above 50°C be of sufficiently short duration so as not to denature the enzymes.

Before, , or after wet milling, a rifying agent is delivered to the tank from a hopper 140, which includes a metering device 142. During saccharification, the contents of the tank are mixed, e.g., by one or more jet mixers. In some cases, the jet mixers are operated during wet milling. A jet mixer 144 is ented diagrammatically in FIGS. 2A and 2B; es of suitable jet mixers will be described in detail below, and are also described in US. Serial Nos. 12/782,694 filed May 18, 2010; I3/293,985 filed November 10, 2011; and 13/293,977 filed November 10, 2011 the full disclosures of . which are hereby incorporated by reference herein. The jet mixer produces a jet using a motor 146 that drives a pump and/or a rotor (not . The torque exerted by the motor 146 correlates with the solids level of the mixture in the tank, which in turn reflects the degree to which the mixture has saccharified. The torque is measured by a torque monitor 148, which sends a signal to a motor 150 that drives the conveyor I30 and also to the metering device 142 of the hopper 140. Thus, the supply of the treated feedstock and the enzyme can be interrupted and resumed as a function of the saccharification of the contents of the tank. The data measured by the torque monitor can also be used to adjust thejet mixer, e.g., to a lower RPM for a mixer that utilizes a rotor, or to a et velocity for a riven mixer. Instead of, or in on to, the torque monitor, the system may include an Amp monitor (not shown) that measures the full load amperage of the motor. In some cases, the jet mixer may include a variable frequency drive (VFD) to allow the speed of the motor to be adjusted.

The system may also include a heat monitor (not shown) that monitors the temperature of the liquid medium and adjusts the feed rate of the feedstock and/or the mixing conditions in se to increases in temperature. Such a temperature feedback loop can be used to prevent the liquid medium from reaching a temperature that will denature the . The heat monitor can also be used to determine when to shut off the wet milling unit to avoid ating of the tank contents.

When one or more pumps are used in the systems described herein, it is generally preferred that positive displacement (PD) pumps be used, e.g., progressive cavity or screw-type PD pumps.- .

In some cases, the cturing plant can be, for example, an existing grain- based or sugar—based l plant or one that has been retrofitted by removing or decommissioning the equipment upstream from the bio-processing system (which in a typical ethanol plant generally includes grain receiving equipment, a hammerrnill, a slurry mixer, cooking equipment and liquefaction equipment). Thus, the feedstock received by the plant is input directly into the tation ent. A retrofitted plant is shown schematically in The use of an existing grain-based or sugar-based ethanol plant in this manner is described in US. Serial No. 12/704,521, filed ry 11, 2010, the full disclosure of which is incorporated herein by reference.

In some embodiments, rather than transporting the saccharified feedstock (sugar solution) to a separate manufacturing plant, or even a separate tank, the sugar solution is inoculated and ted in the same tank or other vessel used for saccharification.

Fermentation can be completed in the same vessel, or can be started in this manner and then completed during transport as discussed above. Saccharifying and ting in a single tank are described in US. ation Serial No. 12/949,044, Nov 18, 2011, the full disclosure of which is incorporated herein by reference.

Generally, the oxygen level in the fermentation vessel should be controlled, e.g., by monitoring the oxygen level and venting the tank, aerating (e.g., by mixing or sparging in oxygen or mixtures of gases containing oxygen) or ating (e.g., by mixing in or sparging in inert gases such as nitrogen, carbon dioxide, helium and/or argon) the mixture as necessary. In some cases, for example where anaerobic conditions are desirable as discussed previously, the rate of mixing is critical. For example, at times 2012/041382 during the process, no mixing may be desirable so that gases produced during fermentation (e.g., C02, H2 and or methane) can more effectively displace oxygen from the fermentation vessel. It is also desirable to monitor the level of ethanol in the vessel, so that when the ethanol level begins to drop the fermentation process can be stopped, e.g., by heating or the addition of sodium bisulfite. Other methods of stopping tation include adding a peroxide (e. g., peroxy acetic acid or hydrogen peroxide), adding succinic acid or a salt thereof, g the contents of the vessel, or reducing the oxygen sparge rate. Combinations of any two or more of these methods may be used. If fermentation is to be conducted or completed during transport, the transportation vessel (e.g., the tank ofa rail car or tanker truck) can be fitted with a control unit that includes an oxygen monitor and ethanol monitor, and a delivery system for delivering sodium bisulfite (or other tation terminating additive) to the tank and/or a system for ing the parameters in the tank to stop fermentation.

If desired, jet mixing can be utilized during fermentation, and if fermentation is conducted in the same vessel as saccharification the same jet mixing equipment can be utilized. However, in some embodimentsjet mixing is not necessary. For example, if fermentation is conducted during transport the movement of the rail car or tanker truck may provide adequate agitation.

DISPERSING, WET MILLING, AND MIXING s are disclosed herein that include one or more tanks, one or more ors, e.g., one or more jet head agitators, and one or more wet mills. In some instances, all mills and agitator heads are within tanks.

Dispersing The optional dispersing unit 134 may e any type of dispersing equipment that wets the ock with the liquid medium. Many dispersing units include a chamber and a rotor in the chamber positioned such that the ock and liquid medium are drawn towards the rotor axially, and forced outward ly to the periphery of the rotor and thus through the outlet of the unit, in the manner of a centrifugal pump. Depending upon the construction of the dispersing unit, a back-up pump may be required (pump 139, discussed above) to draw the fluid through the dispersing unit at high viscosities. Some dispersing units are constructed to generate very high static fluid pressure within the unit; when such units are used a back-up pump is generally not required.

Example of suitable dispersing systems are disclosed in US. Serial No. 12/949,004, filed November 18, 2010, the full disclosure of which is incorporated herein by nce.

Wet Milling Two examples of wet milling heads for use in wet milling unit 160 are shown in FIGS. 4-10 and FIGS. 1 l—17. Each head includes a rotor and a stator, and is mounted on a shaft (not shown) as is well known in the stator mixer art. In both cases, when the rotor and stator are assembled, the gaps between the teeth of the rotor are out of alignment with the gaps between the teeth of the stator. This creates a shearing gap through which liquid flows under high shear during rotation of the rotor.

Wet milling devices are commercially available, for example, from Quadro ering, (Waterloo Ontario), IKA Works Inc., (Wilmington, Delaware), Admix Inc.

(Manchester, NH) and Silverson, (Dartmouth Massachusetts).

In the implementation shown in FIGS. 4-10, the stator includes two tric rings of teeth (see . Under a given set of conditions, this stator ration will generally produce higher shear than the single ring stator configuration shown in FIGS. .20 1 l—17.’ On the other hand, the rotor of the head shown in FIGS. 11-17 includes an impeller-like portion, as will be shown below, which provides a pumping action which may be ble in certain cases.

Referring to FIGS. 4-10, head 162 includes a rotor 164 and a stator 166. The rotor and stator each e a central hub 158 and 159, respectively, which define apertures dimensioned to receive a shaft (not . The shaft is connected to a motor for rotation of the rotor within the stator, with the aperture in the rotor being keyed with the shaft and the shaft rotating freely within the aperture in the stator as is well known in the art.

Arms 161 and 163, respectively, extend from the hubs to support rotor and stator toothed rings. As shown in FIGS. 6 and 8, the stator 166 includes two rings of teeth — an outer ring of teeth 170 and an inner ring ofteeth 171. The rotor 164 includes a single ring of teeth 169, which fit between the rings of the stator in a nested relationship. The upper surface 165 of rotor 164 includes three projections 167 which create turbulence around the head.

As shown in , a clearance 01 is provided between the outer surface 168 of the teeth 169 of the rotor (the OD of the rotor) and the adjacent inner e 172 of the outer ring of teeth 170 of the . nce (1 is preferably small, to generate high shear, and may be, for example, from about 0.01 to 0.250 inch (0.25 to 0.64 mm), e.g., from about 0.03 to 0.10 inch (0.76 to 2.5 mm). The distance between the inner and outer rings of the stator is equal to this clearance plus the radial thickness of the teeth of the rotor, discussed below.

The outer ers of the rotor and stator (ODl and OD2, FIGS. 7 and 8) will depend on the volume of the tank in which the milling head is used, and how many milling heads are positioned in the tank. The outer diameter of the stator, OD2, can be, for example, from about 3 to 50”, e.g., from about 5 to 25 inches, with larger heads being used in larger tanks. As an example, a 4” stator may be used in a 300 gallon tank.

As shown in each tooth 170 on the outer ring of the stator includes a chamfer 174 between its top surface 176 and outer side wall 178.

The circumferential spacing between adjacent teeth is generally the same for the rotor (SI, and both rings of the stator (S2 and S3, ). Like the clearance or, this spacing will also affect the amount of shear generated by the head during rotation of the stator, with a larger spacing resulting in reduced shear. In some entations, the spacing SI, 82 and S3 is about 0.1 to 0.5 inch (2.5 to 12.5 mm).

The tooth size may vary to some extent based on the desired head er, with larger heads having in some cases somewhat larger teeth for durability. However, generally the tooth size and tooth spacing will remain relatively constant as head diameter ses, with the number of teeth increasing with increasing head diameter.

Referring to FIGS. 9 and 10, in some implementations the tooth dimensions can be, for example, as follows: Tl: 0.10” T2: 0.3 5” T3: 0.10” T4: 0.30” T5: 0.10” T6: 0.30” where T1 is the radial thickness of the rotor teeth, T2 is the circumferential thickness of the rotor teeth, T3 is the radial ess of the outer stator teeth, T4 is the circumferential thickness of the outer stator teeth, and T5 and T6 are, respectively, the radial and circumferential thicknesses of the inner stator teeth.

As noted above, an alternative embodiment is shown in FIGS. 1 1-17, in which the stator has only a single row of teeth. This embodiment also differs from that shown in FIGS. 4—10 in other respects.

First, the arms 1 161 of the rotor are curved in two planes, as shown in FIGS. 1 1- 13, causing the rotor to act as an impeller in addition to its shearing action in the rotor/stator arrangement. This impeller functionality is enhanced by the presence of three larger teeth 1180 (see FIGS. 13 and 14) in the rotor ring, which act as extensions of the rotor arms.

Second, the adjacent side walls 1182 of the teeth 1 169 of the rotor are not arranged at an angle R with respect to the radii of the ring, as best seen in FIGS. 14 and 16. This angle may be, for example, from about 0 to 30 degrees. The angle of the teeth helps to pump al through the gap.

The dimensions of the rotor and stator in this embodiment are generally the same as those described above for the ment shown in FIGS. 11-17.

The rotor or stator can be made with a y of materials. For example, ceramics (e.g., oxides, carbides or nitrides), stainless steel, or super alloys (e.g., loy, Inconel, Waspaloy, Ren alloys, Haynes alloys, TMS alloys and CMSX single crystal alloys).

The rotor/stator head is in some cases interchangeable with the jet mixing heads described below, in particular those shown in FIGS. l9-19B. For example, in the case of converting a jet mixer to a rotor/stator, shroud 208 () and mixing element 206 (A) are removed and the rotor/stator head is mounted on shaft 204. shows a cross-sectional view ofa system for wet g that includes a tank (252), two motors (250) two shafts (254), a wet g head (256) and ajet mixer head (258). As shown, one of the shafts is connected to one of the motors on one end and WO 70707 a wet milling head, as described above. Also as shown, the other shaft is connected to the other motor on one end and a jet g head on the other end.

Jet Mixing Particularly advantageous mixers for use during saccharification and fermentation are known as “jet mixers.” In general, le mixers have in common that they produce high velocity circulating flow, for example flow in a toroidal or elliptical pattern.

Generally, preferred mixers exhibit a high bulk flow rate. Preferred mixers provide this mixing action with relatively low energy consumption. It is also generally preferred that 1O the mixer produce vely low shear and avoid heating of the liquid medium, as shear and/or heat can deleteriously affect the saccharifying agent (or microorganism, e.g., in the case of tation). As will be discussed in detail below, some preferred mixers draw the mixture through an inlet into a mixing element, which may include a rotor or impeller, and then expel the mixture from the mixing element through an outlet nozzle.

This ating action, and the high velocity of the jet exiting the nozzle, assist in dispersing material that is floating on the surface of the liquid or material that has settled to the bottom of the tank, depending on the orientation of the mixing element. Mixing elements can be positioned in different orientations to disperse both floating and settling material, and the orientation of the mixing ts can in some cases be adjustable.

In some preferred mixing systems the velocity vo of the jet as meets the ambient fluid is from about 2 to 300 m/s, e.g., about 5 to 150 m/s or about 10 to 100 rn/s. The power consumption of the mixing system may be about 20 to 1000 KW, e.g., 30 to 570 KW, 50 to 500 KW, or 150 to 250KW for a 100,000 L tank.

Jet mixing involves the discharge of a ged jet, or a number of submerged jets, of high velocity liquid into a fluid medium, in this case the e of biomass feedstock, liquid medium and saccharifying agent. The jet of liquid penetrates the fluid medium, with its energy being dissipated by ence and some initial heat. This turbulence is associated with velocity gradients (fluid shear). The surrounding fluid is accelerated and entrained into the jet flow, with this ary entrained flow increasing as the distance from the jet nozzle increases. The momentum of the secondary flow remains generally constant as the jet expands, as long as the flow does not hit a wall, floor or other obstacle. The longer the flow continues before it hits any obstacle, the more liquid is entrained into the secondary flow, increasing the bulk flow in the tank or vessel. When it ters an obstacle, the secondary flow will lose momentum, more or less depending on the geometry ofthe tank, e.g., the angle at which the flow impinges on the obstacle. It is generally ble to orient thejets and/or design the tank so that hydraulic losses to the tank walls are minimized. For example, it may be desirable for the tank to have an arcuate bottom (e.g., a domed headplate), and for the jet mixers to be oriented relatively close to the sidewalls. The tank bottom (lower head plate) may have any desired domed configuration, or may have an elliptical or conical geometry.

Jet mixing s from most types ofliquid/liquid and liquid/solid mixing in that the driving force is hydraulic rather than mechanical. Instead of shearing fluid and propelling it around the mixing vessel, as a mechanical agitator does, ajet mixer forces fluid through one or more nozzles within the tank, creating high—velocity jets that entrain other fluid. The result is shear (fluid against fluid) and circulation, which mix~ the tank contents ntly.

Referring to , the high velocity gradient n the core flow from a submerged jet and the surrounding fluid causes eddies. A illustrates the general characteristics of a submerged jet. As the submerged jet expands into the surrounding t environment the velocity profile flattens as the distance (x) from the nozzle increases. Also, the velocity gradient dv/dr changes with r (the distance from the centerline of the jet) at a given distance x, such that eddies are created which define the ‘ mixing zone (the conical expansion from the nozzle).

In an experimental study ofa submerged jet in air (the results ofwhich are ‘ applicable to any fluid, ing water), Albertson et al. (“Diffusion of Submerged Jets,” Paper 2409, Amer. Soc. of Civil Engineers Transactions, Vol. 1152639-697, 1950, at p. 657) developed dimensionless relationships for V(X]r=0/Vo (centerline velocity), v(x}r=o (velocity profile at a given x), Qx/Qo (flow entrainment), and Ex/Eo (energy change with x): (1) Centerline velocity, v(x) r=o/vo: 2012/041382 v(r = 0) i = 6.2 0 Do [2) velocity profile at any x, v(rjx/v(x]r=o: mgr“) -' i] = 0.79 — 33’—v D (3) Flow and energy at any x: = ‘3 Q0 D, (10.21) g: = 4.11:0 (10.22) where: v(r = 0] = centerline velocity erged jet (m/s), v0 = velocity ofjet as it emerges from the nozzle (m/s), x = distance from nozzle (m), r = distance from centerline ofjet (m), DD = diameter ofnozzle (m), Q = flow of fluid across any given plane at distance x from the nozzle (me/s), Q0 ; flow of fluid emerging from the nozzle (m3/s), E = energy flux of fluid across any given plane at distance x from the nozzle (m3/s), E0 = energy flux of fluid emerging from the nozzle .

(“Water Treatment Unit Processes: Physical and Chemical,” David W. Hendricks, CRC Press 2006, p. 411.) WO 70707 Jet mixing is particularly cost-effective in large-volume (over 1,000 gal) and low— viscosity (under 1,000 cPs) applications. It is also generally advantageous that in most cases the pump or motor of the jet mixer not be submerged, e.g., when a pump is used it is generally located outside the .

One advantage ofjet mixing is that the temperature of the ambient fluid (other than directly adjacent the exit of the nozzle, where there may be some zed g) is increased only slightly if at all. For example, the temperature may be sed by less than 5°C, less than 1°C, or not to any measureable extent.

Jet-Flow Agitators One type ofjet-flow agitator is shown in FIGS. 19-19A. This type of mixer is available commercially, e.g., from IKA under the ame ROTOTRONTM. Referring to , the mixer 200 includes a motor 202, which rotates a drive shaft 204. A mixing element 206 is mounted at the end ofthe drive shaft 204. As shown in A, the mixing t 206 includes a shroud 208 and, within the shroud, an impeller 210.

As indicated by the arrows, when the impeller is rotated in its “forward” direction, the impeller 210 draws liquid in through the open upper end 212 of the shroud and forces the liquid out through the open lower end 214. Liquid exiting end 214 is in the form ofa high velocity stream orjet. 1f the direction of rotation of the impeller 210 is reversed, liquid can be drawn in through the lower end 214 and d through the upper end 212.

This can be used, for example, to suck in solids that are floating near or on the surface of the liquid in a tank or vessel. (It is noted that “upper” and “lower” refer to the orientation of the mixer in ; the mixer may be oriented in a tank so that the upper end is below the lower end.) The shroud 208 includes flared areas 216 and 218 adjacent its ends. These flared areas are believed to contribute to the generally toroidal flow that is observed with this type of mixer. The geometry ofthe shroud and impeller also concentrate the flow into a high velocity stream using relatively low power consumption.

Preferably, the clearance between the shroud 208 and the impeller 210 is sufficient so as to avoid excessive g of the material as it passes through the shroud.

For example, the clearance may be at least 10 times the e‘particle size of the solids in the mixture, preferably at least 100 times.

In some implementations, the shaft 204 is configured to allow gas delivery through the shaft. For example, the shaft 204 may include a bore (not shown) through which gas is delivered, and one or more s through which gas exits into the mixture.

The orifices may be within the shroud 208, to enhance mixing, and/or at other locations along the length of the shaft 204.

The er 210 may have any desired geometry that will draw liquid through the shroud at a high velocity. The impeller is preferably a marine impeller, as shown in A, but may have a different design, for e, a Rushton impeller as shown in B, or a modified Rushton impeller, e.g., tilted so as to provide some axial flow.

In order to generate the high ty flow through the shroud, the motor 202 is preferably a high speed, high torque motor, e.g., capable of operating at 500 to 20,000 RPM, e.g., 3,000 to 10,000 RPM. However, the larger the mixer (e.g., the larger the shroud and/or the larger the motor) the lower the rotational speed can be. Thus, if a large mixer is used, such as a 5 hp, 10 hp, 20 hp, or 30 hp or greater, the motor may be designed to e at lower rotational speeds, e.g., less than 2000 RPM, less than 1500 RPM, or even 500 RPM or less. For example, a mixer sized to mix a 10,000—20,000 liter tank may operate at speeds of 900 to 1,200 RPM. The torque of the motor is ably self-adjusting, to maintain a relatively constant er speed as the mixing conditions changeover time, e.g., due to saccharification of the solids.

Advantageously, the mixer can be oriented at any desired angle.or on in the tank, to direct the jet flow in a desired direction. Moreover, as discussed above, depending on the direction of rotation of the impeller the mixer can be used to draw fluid from either end of the shroud.

In some entations, two or more jet mixers are positioned in the vessel, with one or more being configured to jet fluid upward (“up pump”) and one or more being-configured to jet fluid downward (“down pump”). In some cases, an up pumping mixer will be positioned adjacent a down pumping mixer, to enhance the turbulent flow created by the mixers. If desired, one or more mixers may be switched between upward flow and downward flow during processing. It may be advantageous to switch all or 2012/041382 most of the mixers to up pumping mode during initial dispersion of the feedstock in the liquid medium, particularly if the feedstock is dumped or blown onto the surface of the liquid, as up pumping creates significant turbulence at the surface. Up pumping can also be used during fermentation to help remove CO; from the liquid by causing the gas to bubble to the e where it can be vented.

Other suitable jet mixers are described in US. Applications Serial Nos. 12/782,694 filed May 18,2011; 13/293,985 filed November 10, 2011; 13/293,977 filed November 10, 2011 and US. 12/782,694, filed May 18, 2010, the full disclosures of which are incorporated herein by reference.

MATERIALS Biomass Materials The biomass can be, e.g., a cellulosic or lignocellulosic material. Such materials include paper and paper products (e.g., polycoated paper and Kraft paper), wood, wood- related materials, e.g., particle board, grasses, rice hulls, bagasse, jute, hemp, flax, bamboo, sisal, abaca, straw, switchgrass, alfalfa, hay, corn cobs, corn stover, wheat straw, coconut hair; and materials high in a—cellulose content, e.g., cotton. Feedstocks can be obtained from virgin scrap textile als, e. g., remnants, post consumer waste, e. g., rags. When paper ts are used they can be virgin materials, e.g., scrap virgin als, or they can be post—consumer waste. Aside from virgin raw materials, post- er, industrial (e.g., offal), and sing waste (e.g., effluent from paper processing) can also be used as fiber sources. s feedstocks can also be obtained or d from human (e. g., sewage), animal or plant wastes. Additional cellulosic and lignocellulosic materials have been described in US. Patent Nos. 6,448,307; 6,258,876;6,207,729; 5,973,035 and 5,952,105.

In some embodiments,'the biomass material includes a carbohydrate that is or includes a material having one or more B— l ,4-linkages and having a number average molecular weight between about 3,000 and . Such a carbohydrate is or includes cellulose (I), which is derived from cose 1) through condensation of B(l,4)- glycosidic bonds. This linkage contrasts itself with that for a(1,4)-glycosidic bonds present in starch and other carbohydrates.

HO OH In some ments, the biomass material includes starchy materials, e. g., corn starch, wheat starch, potato starch or rice , a derivative of starch, or a material that es starch, such as an edible food product or a crop. For example, the starchy material can be arracacha, buckwheat, banana, barley, corn kernels, cassava, kudzu, oca, sago, sorghum, regular household potatoes, sweet potato, taro, yams, or one or more beans, such as favas, lentils or peas. Blends of any two or more starchy materials are also starchy materials. Mixtures of starchy, cellulosic and or lignocellulosic materials can also be used. For example, a biomass can be an entire plan, a part of a plant or different parts of a plant e.g., a wheat plant, cotton plant, a corn plant, rice plant or a tree. The starchy als can be treated by any of the s described herein. 2012/041382 In other embodiments, the biomass materials, such as cellulosic, starchy and lignocellulosic feedstock materials, can be obtained from plants that have been modified with respect to a wild type variety. Such ations may be, for example, through the iterative steps of selection and breeding to obtain desired traits in a plant. Furthermore, the plants can have had genetic material removed, modified, silenced and/or added with respect to the wild type variety. For example, genetically modified plants can be produced by recombinant DNA methods, where genetic modifications include introducing or modifying specific genes from parental varieties, or, for e, by using transgenic breeding wherein a specific gene or genes are introduced to a plant from a different species of plant and/or bacteria. Another way to create genetic variation is through mutation breeding wherein new s are artificially created from neous genes. The artificial genes can be created by a y of ways including treating the plant or seeds with, for example, chemical mutagens (e.g., using alkylating agents, epoxides, alkaloids, peroxides, dehyde), irradiation (e.g., X—rays, gamma rays, neutrons, beta particles, alpha particles, protons, deuterons, UV radiation) and temperature shocking or other external stressing and subsequent selection techniques. Other methods of providing modified genes is through error prone PCR and DNA shuffling followed by insertion of the desired modified DNA into the desired plant or seed. Methods of introducing the desired genetic variation in the seed or plant include, for example, the use of a ial carrier, biolistics, calcium phosphate itation, electroporation, gene splicing, gene silencing, lipofection, microinjection and viral carriers. Additional genetically modified materials have been described in US. ation Serial No 13/396,369 filed February 14, 2012 the full disclosure of which is orated herein by reference.

In some cases the biomass is a microbial material. Microbial sources include, but are not limited to, any naturally occurring or genetically d rganism or organism that contains or is capable of providing a source of ydrates (e. g., cellulose), for example, protists, e.g., animal protists (e.g., protozoa such as flagel-lates, amoeboids, es, and oa) and plant protists (e.g., algae such alveolates, chlorarachniophytes, cryptomonads, euglenids, glaucophytes, haptophytes, red algae, stramenopiles, and viridaeplantae). Other examples include seaweed, on (e. g., macroplankton, mesoplankton, microplankton, nanoplankton, picoplankton', and femptoplankton), phytoplankton, bacteria (e.g., gram positive bacteria, gram negative ia, and extremophiles), yeast and/or mixtures of these. In some instances, ial biomass can be obtained from natural sources, e.g., the ocean, lakes, bodies of water, e.g., salt water or fresh water, or on land. Alternatively or in addition, microbial s can be obtained from culture systems, e.g., large scale dry and wet culture systems. rifying Agents Suitable enzymes include cellobiases, cellulases and amylases capable of degrading 1O biomass.

Suitable cellobiases include a cellobiase from Aspergillus niger sold under the tradename NOVOZYME 188W.

Cellulases are capable of degrading s, and may be of fungal or bacterial origin. Suitable s include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, Chrysosporium and Trichoderma, and include species of Humicola, Caprinus, Thielavia, Fusarium, Myceliophthora, Acremonium, Cephalosporium, Scytalidium, Penicillium or Aspergillus (see, e. g., EP 458162), especially those produced by a strain selected from the species Humicola insolens (reclassified as Scytalidium thermophilum, see, e.g, US Patent No. 4,435,307), Coprinus us, Fusarium oxysporum, Myceliophthora thermophi/a, Meripilus giganteus, via terrestris, Acremom'um sp., Acremonium persicinum, nium acremonium, Acremonium brachypenium, Acremonium dichromosporum, Acremonium obclavatum, Acremonium pinkertoniae, Acremonium roseogriseum, Acremonium ratum, and niumfuraium; preferably from the s la insolens DSM 1800, Fusarium oxysporum DSM 2672, Myceliophthora thermophi/a CBS 1 17.65, Cephalosporium sp. RYM-202, Acremonium Sp. CBS 478.94, Acremonium Sp. CBS 265.95, Acremonium persicinum CBS 169.65, Acremonium acremom'um AHU 9519, osporium sp. CBS 535.71, Acremonium peniuin CBS 866.73, Acremonium dichromosporum CBS 683.73, Acremom'um obclavatum CBS 31 1.74, Acremonium pinkertoniae CBS 157.70, Acremonium roseogriseum CBS 134.56, Acremonium incoloratum CBS 146.62, and Acremoniumfuratum CBS 299.70H. Cellulolytic enzymes may also be obtained from Chrysosporium, preferably a strain of Chrysosporium wense. Additionally, Trichoderma (particularly Trichoderma viride, Trichoderma reesei, and Trichoderma koningii), alkalophilic Bacillus (see, for example, US. Patent No. 3,844,890 and EP 458162), and omyces (see, e.g., EP 458162) may be used.

Enzyme complexes may be utilized, such as those available from Genencore under the tradename ACCELLERASE®, for example, Accellerase® 1500 enzyme complex. erase® 1500 enzyme complex contains multiple enzyme activities, mainly exoglucanase, endoglucanase (2200-2800 CMC U/g), hemi-cellulase, and beta- glucosidase (525-775 pNPG U/g), and has a pH of 4.6 to 5.0. The endoglucanase activity of the enzyme x is expressed in carboxymethylcellulose activity units (CMC U),_ while the beta—glucosidase activity is ed in pNP-glucoside activity units (pNPG U).

In one embodiment, a blend of Accellerase® 1500 enzyme complex and NOVOZYMETM 188 cellobiase is used.

In some implementations, the saccharifying agent comprises an acid, e.g., a mineral acid. When an acid is used, co-products may be generated that are toxic to microorganisms, in which case the process can further include removing such co- products. Removal may be performed using an activated carbon, e.g., ted charcoal, or other le techniques.

Fermentation Agents The microorganism(s) used in fermentation can be natural microorganisms and/or engineered microorganisms. For example, the microorganism can be a bacterium, e. g., a cellulolytic bacterium, a fungus, e.g., a yeast, a plant or a protist, e.g., an algae, a protozoa or a fungus-like protist, e. g., a slime mold. When the organisms are compatible, mixtures of sms can be utilized.

Suitable fermenting microorganisms have the ability to convert carbohydrates, such as glucose, xylose, arabinose, e, galactose, accharides or ccharides into fermentation products. Fermenting microorganisms include strains of the genus Sacchromyces Spp. e.g., Sacchromyces cerevisiae (baker’s yeast), Saccharomyces distaticus, Saccharomyces uvarum; the genus Kluyveromyces, e. g., species Kluyveromyces marxianus, romycesfragilis; the genus Candida, e.g., Candida pseudotrapicalis, and Candida brassicae, Pichia stipitis (a relative of Candida shehatae, the genus Clavispora, e.g., species Clavispora niae and Clavispora opuntiae, the genus Pachysolen, e.g., species Pachysolen tannophilus, the genus Bretannomyces, e.g., species Bretannomyces clausenii ppidis, G. P., 1996, Cellulose bioconversion technology, in Handbook on Bioethanol: Production and Utilization, Wyman, C.E., ed., Taylor & Francis, Washington, DC, 179-212). Other suitable rganisms include, for example, nas mobilis, Clostridium thermocellum (Philippidis, 1996, supra), Clostridium robugllacetanicum, Clostridium saccharobutylicum, Clostridium Puniceum, Clostridium beijernckii, Clostridium acetobutylicum, Moniliella pollinis, Yarrowia tica, Aureobasidium sp., Trichosporonoides sp., Trigonopsis variabilis, Trichosporon sp., Moniliellaacetoabutans, Typhula variabilis, Candida magnoliae, Ustilaginomycetes, Pseudozyma aensis, yeast species of genera Zygosaccharomyces, Debaryomyces, Hansenula and Pichia, and fungi ofthe dematioid genus Torula. cially ble yeasts include, for example, Red Star®/Lesaffre Ethanol Red (available from Red Star/Lesaffre, USA), FALI® (available from Fleischmann’s Yeast, a division of Burns Philip Food Inc, USA), SUPERSTART® (available from Alltech, now Lalemand), GERT STRAND® (available from Gert Strand AB, Sweden) and FERMOL® (available from DSM Specialties).

ADDITIVES Antibiotics While it is generally preferred to have a high sugar concentration in the saccharified solution, lower concentrations may be used, in which case it may be desirable to add an antimicrobial additive, e.g., a broad spectrum otic, in a low concentration, e.g., 50 to 150 ppm. Other suitable antibiotics include amphotericin B, ampicillin, chloramphenicol, ciprofloxacin, icin, hygromycin B, kanamycin, ,neomycin, penicillin, puromycin, streptomycin. Antibiotics will inhibit growth of microorganisms during transport and storage, and can be used at appropriate concentrations, e.g., n 15 and 1000 ppm by weight, e.g., between 25 and 500 ppm, or n 50 and 150 ppm. If desired, an antibiotic can be included even ifthe sugar concentration is relatively high.

Surfactants The addition of surfactants can enhance the rate of saccharification. Examples of surfactants include non-ionic surfactants, such as a Tween® 20 or Tween® 80 polyethylene glycol surfactants, ionic surfactants, or amphoteric surfactants. Other suitable surfactants include octylphenol ethoxylates such as the TRITONTM X series nonionic tants commercially available from Dow Chemical. A tant can also be added to keep the sugar that is being produced in solution, particularly in high concentration solutions.

Saccharification Medium In one embodiment, the medium has the following concentrations of components: PHYSICAL TREATMENT OF FEEDSTOCK Physical Preparation In some cases, methods can include a physical preparation, e.g., size reduction of materials, such as by cutting, grinding, shearing, izing or chopping. For example, in other cases, material is first ated or processed using one or more of the methods described herein, such as radiation, sonication, oxidation, pyrolysis or steam explosion, and then size reduced or further size reduced. Treating first and then size reducing can be advantageous. Screens and/or magnets can be used to remove oversized or undesirable objects such as, for e, rocks or nails from the feed stream. In some cases no pre- processing is necessary, for example when the initial itrance of the biomass is low, and wet milling is sufficiently effective to reduce the recalcitrance, for example, to prepared the material for further processing, e.g., saccharification.

Feed preparation systems can be configured to produce s with specific characteristics such as, for example, specific maximum sizes, specific length-to-width,or c surface areas ratios. Physical preparation can increase the rate of reactions or reduce the processing time ed by opening up the materials and making them more accessible to processes and/or reagents, such as reagents in a solution. The bulk density of feedstocks can be controlled (e. g., increased). In some situations, it can be desirable to prepare a high or higher bulk density material, e.g., by densifying the material (e. g., densification can make it easier and less costly to transport to another site) and then reverting the material to a lower bulk y state. The material can be densified, for example from less than 0.2 g/cc to more than 0.9 g/cc (e.g., less than 0.3 to more than 0.5 g/cc, less than 0.3 to more than 0.9 g/cc, less than 0.5 to more than 0.9 g/cc, less than 0.3 to more than 0.8 g/cc, less than 0.2 to more than 0.5 g/cc). For example, the material can be densified by the methods and equipment disclosed in US. 7,932,065 and WO 2008/073186, the full disclosures of which are incorporated herein by reference.

Densified materials can be processed by any of the s described herein, or any material processed by any of the methods described herein can be subsequently densified.

In some cases, the material can be densified prior to wet g. Wet milling can re-open densified material.

Size Reduction In some embodiments, the al to be sed is in the form of a fibrous al that includes fibers provided by shearing a fiber . For e, the shearing can be performed with a rotary knife cutter.

For example, a fiber source, e.g., that is recalcitrant or that has had its recalcitrance level reduced, can be sheared, e.g., in a rotary knife , to provide a first fibrous material. The first fibrous material is passed through a first screen, e.g., having an average opening size of 1.59 mm or less (1/16 inch, 0.0625 inch), provide a second fibrous material. If desired, the fiber source can be cut prior to the shearing, e.g., with a shredder. For example, when a paper is used as the fiber source, the paper can be first cut WO 70707 into strips that are, e.g., 1/4— to l/2-inch wide, using a shredder, e.g., a counter-rotating screw er, such as those manufactured by Munson (Utica, NY). As an alternative to shredding, the paper can be reduced in size by cutting to a desired size using a guillotine cutter. For example, the guillotine cutter can be used to cut the paper into sheets that are, e.g., 10 inches wide by 12 inches long.

In some ments, the shearing of the fiber source and the passing of the ing first fibrous material through a first screen are performed concurrently. The shearing and the passing can also be performed in a batch-type process.

For example, a rotary knife cutter can be used to concurrently shear the fiber source and screen the first fibrous material. A rotary knife cutter includes a hopper that can be loaded with a shredded fiber source prepared by shredding a fiber source. The shredded fiber source In some implementations, the feedstock is physically treated prior to saccharification and/or fermentation. Physical treatment processes can include one or more of any of those described herein, such as mechanical treatment, chemical treatment, irradiation, tion, oxidation, pyrolysis or steam explosion. Treatment methods can be used in combinations of two, three, four, or even all of these logies (in any order). When more than one treatment method is used, the methods can be applied at the same time or at different times. Other processes that change a molecular structure of a biomass feedstock may also be used, alone or in combination with the processes disclosed herein.

Mechanical Treatments In some cases, s can include mechanically treating the biomass ock.

Mechanical treatments include, for example, cutting, milling, ng, grinding, shearing and chopping. Milling may include, for example, ball milling, hammer milling, rotor/stator dry or wet milling, r milling, blade g, knife milling, disk milling, roller milling or other types of g. Other mechanical treatments include, e.g., stone grinding, cracking, mechanical ripping or tearing, pin grinding or air attrition milling.

Mechanical treatment can be advantageous for “opening up,9’ (Sstressing,” breaking and shattering the cellulosic or lignocellulosic materials, making the cellulose of the materials more susceptible to chain scission and/0r ion of crystallinity. The open materials can also be more susceptible to oxidation when ated.

In some cases, the mechanical treatment may include an initial preparation of the feedstock as received, e.g., size reduction of materials, such as by cutting, grinding, shearing, pulverizing or chopping. For example, in some cases, loose feedstock (e.g., recycled paper, starchy materials, or switchgrass) is ed by shearing or shredding.

Alternatively, or in addition, the feedstock material can first be physically treated by one or more of the other physical treatment methods, e.g., chemical treatment, radiation, sonication, ion, pyrolysis or steam explosion, and then mechanically treated. This sequence can be advantageous since materials treated by one or more of the other treatments, e. g., ation or sis, tend to be more brittle and, therefore, it may be easier to further change the molecular structure of the material by ical treatment.

In some embodiments, the feedstock material is in the form of a fibrous material, and mechanical ent includes shearing to expose fibers of the fibrous material.

Shearing can be performed, for e, using a rotary knife cutter. Other methods of mechanically ng the feedstock include, for example, milling or grinding. Milling may be performed using, for example, a hammer mill, ball mill, colloid mill, conical or cone mill, disk mill, edge mill, Wiley mill or grist mill. ng may be performed using, for example, a stone grinder, pin grinder, coffee grinder, or burr grinder. Grinding may be provided, for example, by a reciprocating pin or other element, as is the case in a pin mill. Other mechanical treatment methods include mechanical ripping or tearing, other methods that apply pressure to the material, and air attrition milling. Suitable ical treatments further include any other technique that changes the molecular structure of the feedstock.

If desired, the mechanically treated material can be passed through a screen, e.g., having an average opening size of 1.59 mm or less (1/16 inch, 0.0625 inch). In some embodiments, shearing, or other mechanical ent, and screening are performed concurrently. For example, a rotary knife cutter can be used to concurrently shear and screen the feedstock. The feedstock is sheared between stationary blades and rotating blades to provide a sheared material that passes through a screen, and is captured in a bin.

The cellulosic or lignocellulosic material can be mechanically treated in a dry state (e. g., having little or no free water on its surface), a hydrated state (e.g., having up to ten percent by weight absorbed water), or in a wet state, e.g., having between about 10 percent and about 75 percent by weight water. The fiber source can even be mechanically treated while partially or fully submerged under a liquid, such as water, l or isopropanol.

The fiber cellulosic or lignocellulosic material can also be mechanically treated under a gas (such as a stream or atmosphere of gas other than air), e. g., oxygen or nitrogen, or steam.

If desired, lignin can be removed from any of the fibrous materials that include . Also, to aid in the breakdown of the materials that include cellulose, the material can be treated prior to or during mechanical treatment or irradiation with heat, a chemical (e.g., mineral acid, base or a strong oxidizer such as sodium hypochlorite) and/or an enzyme. For example, grinding can be performed in the presence of an acid.

Mechanical treatment systems can be configured to produce streams with c morphology characteristics such as, for example, surface area, porosity, bulk density, and, in the case of fibrous ocks, fiber characteristics such as —to-width ratio.

In some embodiments, a BET surface area of the mechanically treated material is greater than 0.1 mZ/g, e.g., greater than 0.25 mZ/g, r than 0.5 mZ/g, greater than 1.0 mz/g, greater than 1.5 mZ/g, greater than 1.75 mz/g, greater than 5.0 mZ/g, greater than 10 mz/g, greater than 25 mZ/g, r than 35 mz/g, greater than , greater than 60 mz/g, r than 75 mz/g, greater than 100 mZ/g, greater than 150 mZ/g, greater than 200 mz/g, or even greater than 250 mZ/g.

A porosity of the mechanically treated material can be, e.g., greater than 20 percent, greater than 25 percent, greater than 35 percent, greater than 50 percent, greater than 60 percent, greater than 70 percent, greater than 80 percent, greater than 85 percent, greater than 90 percent, greater than 92 percent, greater than 94 percent, greater than 95 percent, greater than 97.5 percent, greater than 99 t, or even greater than 99.5 t. a In some embodiments, after ical treatment the material has a bulk density ofless than 0.25 g/cm3, e.g., 0.20 g/cm3, 0.15 g/cm3, 0.10 g/cm3, 0.05 g/cm3 or less, e.g., 0.025 g/cm3. Bulk density is determined using ASTM D1895B. Briefly, the method involves filling a ing cylinder of known volume with a sample and obtaining a weight of the sample. The bulk density is calculated by dividing the weight of the sample in grams by the known volume of the cylinder in cubic eters.

If the ock is a fibrous material the fibers ofthe fibrous materials ically treated material can have a relatively large average length-to-diameter ratio (e.g., greater than 20-to-1), even if they have been sheared more than once. In on, the fibers of the fibrous materials described herein may have a relatively narrow length and/or length-to—diameter ratio distribution.

As used herein, e fiber widths (e.g., diameters) are those determined optically by randomly selecting approximately 5,000 fibers. e fiber lengths are corrected length—weighted lengths. BET (Brunauer, Emmet and Teller) surface areas are multi-point surface areas, and porosities are those determined by mercury porosimetry.

If the second feedstock is a fibrous material 14 the average length-to-diameter ratio of fibers of the mechanically treated al can be, e.g. greater than 8/1, e.g., r than 10/1, greater than 15/ 1, greater than 20/1, greater than 25/1, or greater than 50/1. An average fiber length of the mechanically treated material 14 can be, e.g., n about 0.5 mm and 2.5 mm, e.g., between about 0.75 mm and 1.0 mm, and an average width (e. g., diameter) of the second fibrous material 14 can be, e.g., between about 5 um and 50 um, e.g., between about 10 um and 30 pm.

In some embodiments, if the feedstock is a fibrous material, the standard deviation of the fiber length of the mechanically treated material can be less than 60 percent of an average fiber length of the mechanically treated material, e.g., less than 50 percent of the average length, less than 40 percent of the average length, less than 25 percent of the e length, less than 10 percent of the average length, less than 5 percent of the average length, or even less than 1 percent of the e length.

Treatment to Solubilize, Reduce Recalcitrance or Functionalize Materials that have or have not been physically prepared can be treated for use in any production process described herein. One or more of the production ses described below may be included in the recalcitrance reducing operating unit discussed above. Alternatively, or in addition, other processes for reducing recalcitrance may be included.

Treatment processes utilized by the recalcitrance reducing operating unit can include one or more of irradiation, sonication, oxidation, pyrolysis or steam explosion.

Treatment methods can be used in combinations of two, three, four, or even all of these technologies (in any order).

Radiation ent One or more radiation processing sequences can be used to process materials from the feedstock, and to provide a wide variety of different sources to extract useful substances from the feedstock, and to provide partially degraded structurally modified material which functions as input to further processing steps and/or sequences.

Irradiation can, for example, reduce the molecular weight and/or crystallinity of feedstock. Radiation can also sterilize the materials, or any media needed to bioprocess the material.

In some ments, energy ted in a material that releases an electron from its atomic orbital is used to irradiate the materials. The radiation may be provided by (1) heavy d particles, such as alpha particles or protons, (2) electrons, produced, for example, in beta decay or electron beam accelerators, or (3) electromagnetic radiation, for example, gamma rays, x rays, or ultraviolet rays. In one ch, radiation produced by radioactive substances can be used to irradiate the feedstock. In some embodiments, any combination in any order or concurrently of(1) through (3) may be utilized. In another approach, electromagnetic radiation (e.g., ed using electron beam emitters) can be used to irradiate the feedstock. The doses applied depend on the desired effect and the particular feedstock.

In some ces when chain scission is desirable and/or polymer chain functionalization is desirable, particles r than electrons, such as protons, helium , argon ions, silicon ions, neon ions, carbon ions, phosphorus ions, oxygen ions or nitrogen ions can be utilized. When ring-opening chain scission is d, positively d particles can be utilized for their Lewis acid properties for enhanced ring- opening chain scission. For example, when maximum oxidation is desired, oxygen ions can be utilized. and when maximum nitration is desired, nitrogen ions can be utilized.

The use of heavy particles and positively charged particles is described in US. 7,931,784, the full disclosure of which is incorporated herein by reference.

In one method, a first material that is or includes ose having a first number average molecular weight (Mm) is irradiated, e.g., by treatment with ionizing radiation (e.g., in the form of gamma radiation, X-ray radiation, 100 nm to 280 nm ultraviolet (UV) light, a beam of electrons or other charged particles) to provide a second material that includes cellulose having a second number average molecular weight (Mm) lower than the first number average molecular weight. The second material (or the first and second material) can be combined with a microorganism (with or without enzyme treatment) that can e the second and/or first material or its constituent sugars or lignin to produce an intermediate or a product, such as those described herein.

Since the second material es cellulose having a reduced molecular weight relative to the first material, and in some instances, a reduced crystallinity as well, the second material is generally more dispersible, swellable and/or e, e.g., in a solution containing a microorganism and/or an . These properties make the second material easier to process and more tible to chemical, enzymatic and/or ical attack relative to the first material, which can greatly improve the tion rate and/or production level of a d t, e. g., ethanol. Radiation can also sterilize the materials or any media needed to bioprocess the material.

In some embodiments, the second material can have a level of oxidation (02) that is higher than the level of oxidation (01) of the first material. A higher level of oxidation of the material can aid in its dispersability, swellability and/or solubility, further enhancing the material’s susceptibility to chemical, tic or biological attack. In some embodiments, to increase the level of the oxidation of the second material relative to the first material, the irradiation is performed under an oxidizing environment, e.g., under a blanket of air or oxygen, producing a second material that is more ed than the first material. For example, the second material can have more hydroxyl groups, aldehyde groups, ketone groups, ester groups or carboxylic acid groups, which can increase its hydrophilicity.

Ionizing Radiation Each form of radiation ionizes the carbon-containing material via particular ctions, as determined by the energy of the radiation. Heavy charged particles primarily ionize matter via Coulomb scattering; furthermore, these interactions produce energetic electrons that may further ionize matter. Alpha particles are identical to the nucleus of a helium atom and are produced by the alpha decay of various radioactive nuclei, such as isotopes of bismuth, polonium, astatine, radon, francium, radium, several actinides, such as actinium, thorium, uranium, neptunium, curium, californium, americium, and plutonium.

When les are utilized, they can be neutral (uncharged), positively charged or negatively charged. When charged, the charged particles can bear a single positive or negative charge, or multiple charges, e.g., one, two, three or even four or more charges.

In ces in which chain scission is desired, positively charged particles may be desirable, in part due to their acidic nature. When particles are utilized, the particles can have the mass ofa resting electron, or r, e.g., 500, 1000, 1500, 2000, 10,000 or even 0 times the mass ofa resting electron. For example, the particles can have a mass of from about 1 atomic unit to about 150 atomic units, e.g., from about 1 atomic unit to about 50 atomic units, or from about 1 to about 25, e.g., 1, 2, 3‘, 4, 5, 10, 12 or 15 amu. Accelerators used to accelerate the particles can 'be electrostatic DC, electrodynamic DC, RF linear, ic ion linear or continuous wave. For example, cyclotron type accelerators are available from IBA, Belgium, such as the Rhodotron® system, while DC type accelerators are ble from RDI, now IBA Industrial, such as the Dynamitron®. Ions and ion accelerators are discussed in uctory Nuclear Physics, Kenneth S. Krane, John Wiley & Sons, Inc. (1988), Krsto Prelec, FIZIKA B 6 (1997) 4, 177—206, Chu, William T., “Overview of Light-Ion Beam Therapy” Columbus-Ohio, ICRU-IAEA g, 18—20 March 2006, Iwata, Y. et al., “Altemating-Phase—Focused IH-DTL for Heavy-Ion Medical Accelerators” Proceedings of EPAC 2006, Edinburgh, nd and Leaner, C.M. et al., “Status ofthe Superconducting ECR Ion Source Venus” Proceedings of EPAC 2000, Vienna, Austria.

In some embodiments, a beam of ons is used as the radiation source. A beam of electrons has the advantages of high dose rates (e.g., 1, 5, or even 10 Mrad per 2012/041382 second), high throughput, less containment, and less confinement equipment. Electrons can also be more efficient at causing chain scission. In addition, electrons having energies of 4-10 MeV can have a ation depth of 5 to 30 mm or more, such as 40 mm. In some cases, multiple on beam devices (e.g., le heads, often referred to as “horns”) are used to deliver multiple doses of electron beam radiation to the material. This high total beam power is usually achieved by utilizing multiple accelerating heads. For e, the electron beam device may include two, four, or more accelerating heads. As one example, the electron beam device may include four accelerating heads, each of which has a beam power of 300 kW, for a total beam power 1O of 1200 kW. The use of multiple heads, each of which has a relatively low beam power, prevents excessive temperature rise in the material, thereby preventing burning of the material, and also increases the uniformity of the dose through the thickness of the layer of material. Irradiating with le heads is disclosed in US. ation Serial No. 13/276,192 filed Oct 18, 201 1, the complete sure of which is incorporated herein by reference. on beams can be generated, e.g., by electrostatic generators, cascade generators, transformer generators, low energy accelerators with a scanning system, low energy accelerators with a linear cathode, linear accelerators, and pulsed accelerators.

Electrons as an ionizing radiation source can be useful, e.g., for relatively thin piles of materials, e.g., less than 0.5 inch, e.g., less than 0.4 inch, 0.3 inch, 0.2 inch, or less than 0.1 inch. In some embodiments, the energy of each electron of the electron beam is from about 0.3 MeV to about 2.0 MeV (million electron volts), e.g., from about 0.5 MeV to about 1.5 MeV, or from about 0.7 MeV to about 1.25 MeV.

Electron beam irradiation devices may be procured commercially from Ion Beam Applications, Louvain-la-Neuve, Belgium or the Titan Corporation, San Diego, CA.

Typical electron energies can be 1 MeV, 2 MeV, 4.5 MeV, 7.5 MeV, or 10 MeV.

Typical on beam irradiation device power can be 1 kW, 5 kW, 10 kW, 20 kW, 50 kW, 100 kW, 250 kW, or 500 kW. The level of depolymerization of the feedstock depends on the electron energy used and the dose applied, while exposure time depends on the power and dose. Typical doses may take values of 1 kGy, 5 kGy, lO kGy, 20 kGy, 50 kGy, 100 kGy, or 200 kGy. In a some embodiments energies between 0.25-10 MeV (e.g., 0.5—0.8 MeV, 0.5-5 MeV, 0.8-4 MeV, 0.8-3 MeV, 0.8-2 MeV 0r 0.8-1.5 MeV) can be used.

Electromagnetic Radiation In embodiments in which the irradiating is performed with electromagnetic radiation, the electromagnetic radiation can have, e.g., energy per photon (in electron volts) of greater than 102 eV, e.g., greater than 103, 104, 105, 106, or even greater than 107 eV. In some embodiments, the electromagnetic radiation has energy per photon of between 104 and 107, e.g., between 105 and 106 eV. The electromagnetic radiation can have a ncy of, e.g., greater than 10‘6 Hz, greater than 10'7 Hz, 10”, 1019, 1020, or even greater than 1021 Hz. In some embodiments, the electromagnetic radiation has a frequency of between 1018 and 1022 Hz, e.g., between 10'9 to 102' Hz.

Doses In some embodiments, the irradiating (with any radiation source or a combination of sources) is performed until the material receives a dose of at least 0.25 Mrad, e.g., at least 1.0, 2.5, 5.0, 8.0, 10, 15, 20, 25, 30, 35, 40, 50, or even at least 100 Mrad. In some embodiments, the irradiating is performed until the material receives a dose of between 1.0 Mrad and 6.0 Mrad, e.g., between 1.5 Mrad and 4.0 Mrad, 2 Mrad and 10 Mrad, 5 Mrad and 20 Mrad, 10 Mrad and 30 Mrad, 10 Mrad and 40 Mrad, or 20 Mrad and 50 Mrad.

In some embodiments, the irradiating is performed at a dose rate of between 5.0 and 1500.0 kilorads/hour, e.g., between 10.0 and 750.0 kilorads/hour or between 50.0 and 350.0 ds/hours.

In some embodiments, two or more ion sources are used, such as two or more ionizing radiations. For example, samples can be d, in any order, with a beam of electrons, followed by gamma radiation and UV light having ngths from about 100 nm to about 280 nm. In some embodiments, s are treated with three ionizing radiation sources, such as a beam of electrons, gamma radiation, and energetic UV light.

Sonication, Pyrolysis and Oxidation In addition to ion treatment, the ock may be treated with any one or more of sonication, pyrolysis and oxidation. These treatment processes are described in US. Serial No. 12/417,840 filed April 3, 2009, the disclosure of which is incorporated by nce herein.

Other Processes To Solubiliz_e, Reduce Recalcitrance Or To Functionalize Any of the processes ofthis paragraph can be used alone without any of the processes described herein, or in combination with any of the processes bed herein (in any order): steam explosion, chemical treatment (e.g., acid treatment (including concentrated and dilute acid treatment with mineral acids, such as sulfuric acid, hydrochloric acid and organic acids, such as trifluoroacetic acid), and/or base treatment (e.g., ent with lime or sodium hydroxide», UV treatment, screw extrusion ent, solvent ent (e.g., treatment with ionic liquids) and freeze milling. Some of these processes, for example, are described in US. Series Nos. 12/417,723 filed April 3, 2009; 13/099,151 filed May 2, 2011; and 12/502,629 filed July 14, 2009, the disclosures of which are incorporated herein.

PRODUCTION OF FUELS ACIDS ESTERS AND/OR OTHER PRODUCTS After one or more of the processing steps discussed above have been performed on the biomass, the complex ydrates contained in the cellulose and hemicellulose ons can be processed into fermentable sugars using a saccharification process, as sed above.

After the resulting sugar solution has been transported to a manufacturing facility, the sugars can be converted into a variety of products, such as alcohols, e.g., l, or organic acids. The product obtained depends upon the microorganism utilized and the conditions under which the bioprocessing occurs. These steps can be performed, for example, utilizing the existing equipment of the com-based ethanol manufacturing facility.

The mixing ses and equipment discussed herein may also be used during bioprocessing, if desired. Advantageously, the mixing systems described herein do not impart high shear to the liquid, and do not significantly raise the overall ature of the liquid. As a result, the microorganisms used in bioprocessing are maintained in a viable condition throughout the process. Mixing may enhance the reaction rate and improve the efficiency of the process. lly, fermentation utilizes various microorganisms. The sugar on produced by saccharification of lignocellulosic materials will generally contain xylose as well as glucose. It may be desirable to remove the xylose, e.g., by chromatography, as some commonly used microorganisms (e.g., yeasts) do not act on xylose. The xylose may be collected and utilized in the manufacture of other products, e.g., animal feeds and the sweetener Xylitol. The xylose may be removed prior to or after delivery of the sugar solution to the manufacturing facility where fermentation will be performed.

The microorganism can be a natural rganism or an engineered microorganism, e.g., any ofthe microorganisms discussed in the Materials section herein.

The optimum pH for yeast is from about pH 4 to 5, while the optimum pH for Zymomonas is from about pH 5 to 6. Typical fermentation times are about 24 to 96 hours with temperatures in the range of 26 °C to 40 °C, however thermophilic microorganisms prefer higher temperatures.

Carboxylic acid groups generally lower the pH of the fermentation solution, tending to inhibit fermentation with some microorganisms, such Pichia is.

Accordingly, it is in some cases desirable to add base and/or a , before or during tation, to bring up the pH of the solution. For example, sodium hydroxide or lime can be added to the fermentation medium to elevate the pH of the medium to range that is optimum for the microorganism utilized.

Fermentation is generally ted in an aqueous growth medium, which can contain a nitrogen source or other nutrient source, e.g., urea, along with vitamins and trace minerals and metals. It is generally preferable that the growth medium be sterile, or at least have a low ial load, e.g., bacterial count. Sterilization of the growth medium may be accomplished in any desired manner. However, in preferred implementations, ization is accomplished by irradiating the growth medium or the individual components of the growth medium prior to mixing. The dosage of radiation is generally as low as le while still obtaining adequate results, in order to minimize energy consumption and resulting cost. For e, in many instances, the growth medium itself or components Of the growth medium can be treated with a radiation dose ofless than 5 Mrad, such as less than 4, 3, 2 or 1 Mrad. In ic instances, the growth medium is treated with a dose of between about 1 and 3 Mrad.

In some embodiments, all or a n of the fermentation process can be interrupted before the low molecular weight sugar is completely ted to ethanol. The intermediate fermentation products include high concentrations of sugar and ydrates. These intermediate fermentation products can be used in preparation of food for human or animal consumption. Additionally or alternatively, the intermediate fermentation products can be ground to a fine particle size in a stainless-steel laboratory mill to produce a flour-like nce.

Mobile ntors can be utilized, as described in US. Series No. 12/374,549 filed Jan 21, 2009, now Published ational Application No. .

Similarly, the saccharification equipment can be mobile. Further, saccharification and/or fermentation may be performed in part or ly during transit.

POST-PROCESSING After fermentation, the resulting fluids can be distilled using, for example, a “beer column” to separate ethanol and other alcohols from the majority of water and residual solids. The vapor exiting the beer column can be, e. g., 35% by weight ethanol and can be fed to a rectification column. A mixture of nearly azeotropic (92.5%) ethanol and water from the rectification column can be purified to pure (99.5%) ethanol using phase molecular sieves. The beer column bottoms can be sent to the first effect ofa three-effect ator. The rectification column reflux condenser can provide heat for this first effect. After the first effect, solids can be ted using a centrifuge and dried in a rotary dryer. A portion (25%) of the centrifuge effluent can be recycled to fermentation and the rest sent to the second and third evaporator effects. Most of the evaporator condensate can be returned to the process as fairly clean condensate with a small portion split off to waste water treatment to prevent build-up of low—boiling compounds.

INTERMEDIATES AND PRODUCTS Using the processes described herein, the treated biomass can be converted to one or more products, such as energy, fuels, foods and materials. Specific examples of products include, but are not limited to, hydrogen, sugars (e.g., glucose, , arabinose, mannose, galactose, fructose, disaccharides, oligosaccharides and polysaccharides), alcohols (e.g., monohydric alcohols or dihydric alcohols, such as ethanol, n-propanol, isobutanol, sec-butanol, tert-butanol or n-butanol), hydrated or hydrous alcohols, e.g., ning greater than 10%, 20%, 30% or even greater than 40% water, xylitol, biodiesel, organic acids, hydrocarbons (e.g., methane, ethane, propane, isobutene, pentane, n-hexane, biodiesel, bio-gasoline and mixtures f), co- products (e.g., proteins, such as cellulolytic proteins es) or single cell proteins), and es of any of these in any ation or relative concentration, and optionally in combination with any additives, e.g., fuel additives. Other examples include carboxylic acids, salts ofa carboxylic acid, a e of carboxylic acids and salts of carboxylic acids and esters of carboxylic acids (e.g., methyl, ethyl and yl esters), ketones (e.g., acetone), aldehydes (e.g., acetaldehyde), alpha, beta unsaturated acids, such as acrylic acid and olefins, such as ethylene. Other alcohols and alcohol tives include propanol, propylene glycol, 1,4-butanediol, 1,3-propanedi01, sugar alcohols (e.g., erythritol, , glycerol, sorbitol threitol, arabitol, ribitol, mannitol, dulcitol, fucitol, iditol, isomalt, maltitol, lactitol, xylitol and other polyols), methyl or ethyl esters of any of these ls. Other products include methyl acrylate, methylmethacrylate, lactic acid, citric acid, formic acid, acetic acid, propionic acid, butyric acid, ic acid, c acid, caproic acid, 3-hydroxypropionic acid, palmitic acid, stearic acid, oxalic acid, malonic acid/glutaric acid, oleic acid, linoleic acid, glycolic acid, y-hydroxybutyric acid, and mixture thereof, a salt ofany of these acids, or a mixture of any of the acids and their respective salts. a salt of any ofthe acids and a mixture of any of the acids and respective salts.

Other intermediates and products, including food and pharmaceutical products, are described in US. Serial No. 12/417,900 filed April 3, 2009, the full disclosure of which is hereby incorporated by reference .

OTHER EMBODIMENTS A number of embodiments have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope ofthe disclosure.

In some implementations, the s discussed herein, or components of these systems, may be portable, e.g., in the manner of the mobile processing equipment described in U.S. Serial No. 12/374,549 filed June 2, 2009 and International Application No. WO 11598, the full disclosures of which are incorporated herein by reference.

While tanks have been referred to herein, the process may take place in any type of vessel or container, including lagoons, pools, ponds and the like. If the container in which mixing takes place is an in-ground structure such as a lagoon, it may be lined. The container may be covered, e.g., if it is outdoors, or uncovered.

In an alternate embodiment, the dispersing system 134 can be omitted in the systems shown in FIGS. 2A and 2B, and a pump can be used to draw liquid from the tank and deliver it through outlet pipe 137 to wet the feedstock material, which is then dispersed by the mixing action of the jet mixer 144. In such implementations, the pump would ably be a low shear pump, e.g., a positive displacement pump such as the progressive cavity pumps available from SEEPEX and lobe pumps from Waukesha. It is also preferred that the pump be capable of pumping high viscosity fluids, since the ity of the liquid will se as more feedstock is added.

While biomass feedstocks have been bed , other feedstocks and mixtures of s feedstocks with other feedstocks may be used. For example, some implementations may utilize mixtures of biomass feedstocks with hydrocarbon-containing feedstocks such as those disclosed in U.S. Series No. 13/293,985 filed November 10, 2011, the full disclosure of which is incorporated by reference herein.

Accordingly, other embodiments are within the scope of the following claims.

Throughout the specification and claims, unless the context es otherwise, the word “comprise” or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated r or group of integers but not the exclusion of any other integer or group of integers.

Each document, reference, patent application or patent cited in this text is expressly orated herein in their entirety by reference, which means that it should be read and ered by the reader as part of this text. That the document, reference, patent application or patent cited in this text is not repeated in this text is merely for reasons of conciseness.

Reference to cited material or information contained in the text should not be understood as a concession that the material or information was part of the common general knowledge or was known in Australia or any other country.

Claims (30)

WHAT IS CLAIMED IS:

1. A method for producing a sugar solution from s comprising: treating the biomass to reduce its itrance, thereby making treated biomass, 5 wet milling and simultaneously jet mixing the treated biomass to make wet milled biomass; rifying the wet milled biomass, thereby producing the sugar solution. 10

2. The method according to claim 1 wherein treating the biomass is by irradiating.

3. The method according to claim 2 wherein ating comprises exposing the biomass to an electron beam.

4. The method according to claim 2 or 3 n irradiating comprises delivering a dose of at least 5 Mrad to the biomass.

5. The method according to any one of claims 1-4, wherein the biomass is 20 ed from the group ting of wood, particle board, sawdust, agricultural waste, sewage, silage, grasses, rice hulls, bagasse, cotton, jute, hemp, flax, bamboo, sisal, abaca, straw, wheat straw, corn cobs, corn stover, switchgrass, alfalfa, hay, coconut hair, seaweed, algae, and mixtures thereof.

6. The method according to any one of claims 1-5 further comprising 25 densifying the treated biomass prior to wet milling.

7. The method according to any one of claims 1-6, wherein wet milling is performed using a rotor/stator head.

8. The method according to claim 7 wherein the rotor and stator include 30 nesting rings of teeth.

9. The method ing to claim 8 wherein the stator comprises two or more concentric rings of teeth.

10. The method according to claim 9 wherein the rotor comprises a ring 5 of teeth configured to fit between adjacent rings of teeth of the stator.

11. The method according to claim 7 wherein the clearance n the rotor and stator is from about 0.01 to 0.25 inches (0.25 to 6.4 mm). 10

12. The method according to claim 8 wherein the spacing between the teeth in each ring of teeth is from about 0.1 to 0.3 inch (2.5 to 7.6 mm).

13. The method according to claim 7 wherein wet milling is performed using a plurality of rotor/stator heads.

14. The method according to claim 1 n wet milling is performed at a shear rate of from about 30,000 sec-1 to 50,000 sec-1.

15. The method according to claim 1 wherein wet milling is performed in- 20 line.

16. The method according to claim 1 wherein the jet mixing is by one or more jet heads and the wet milling is by one or more wet milling heads in a tank sing fluid having biomass dispersed therein.

17. The method according to claim 1 further sing adding an enzyme to the treated biomass before, or during wet milling, or to the wet milled biomass after wet milling. 30

18. The method according to claim 17 further comprising adding a microorganism to the treated biomass or to a sugar derived from the wet milled biomass.

19. The method according to claim 18 wherein the microorganism is added after wet milling has been completed. 5

20. The method according to claim 18 wherein the microorganism converts the sugar to a product selected from the group consisting of alcohols, organic acids, sugars, hydrocarbons, and mixtures thereof.

21. The method according to claim 20 where a jet mixer is operated 10 during conversion of the sugar to the t.

22. A wet milling system comprising one or more jet heads and one or more wet milling heads in a tank sing fluid having biomass dispersed therein.

23. The system according to claim 22 wherein the biomass comprises a lignocellulosic material.

24. The system according to claim 23 wherein the itrance of the 20 lignocellulosic material has been d by ating the biomass.

25. The system according to claim 24 wherein irradiating comprises exposing the biomass to an electron beam. 25

26. The system according to any one of claims 22-25, wherein the wet mill is performed using a rotor/stator head.

27. The system according to claim 26, wherein the rotor and stator include g rings of teeth.

28. The system according to claim 27, wherein the stator comprises two or more concentric rings of teeth.

29. The method according to claim 1 substantially as herein before described with reference to the Examples.

30. The system ing to claim 22 substantially as herein before 5 described with reference to the Examples.