JPS5834111B2 - Saccharification and fermentation method for non-cooked grains - Google Patents

Description

[Detailed description of the invention] The present invention relates to a method for saccharification and fermentation of non-cooked grains.

More specifically, powdered or granular grains or crushed grains thereof are soaked in an alkaline solution that has the ability to swell and swell starch and is easy to neutralize and remove, to swell and swell starch, and after neutralization, the saccharifying enzyme is extracted. Alternatively, it relates to a method in which saccharification is performed by adding a saccharifying enzyme preparation or koji (malt), and fermentation is carried out by the action of yeast or the like.

Conventionally, when producing sugars, liquors, alcohol, and various organic acids using grains with high starch content, the grains are usually steamed after absorbing sufficient water, or water is added and pressure steamed. The starch is then saccharified and then saccharified by the action of a saccharifying enzyme to collect the sugars as a product, or it is usually followed by alcoholic fermentation, organic acid fermentation, etc.

The fact that starch is oxidized by dimethyl sulfoxide, urea, or various alkalis is found in the book (for example, "Starch Science Handbook" supervised by Nikoku Nibu, page 35, published by Asakura Shoten in 1982).

However, to date, there have been no reports of research that applies these drugs to starch-containing grains themselves, refrigerates and saccharifies them, and then ferments them to obtain useful substances such as alcohol or organic acids.

Recently, the inventors of the present invention revealed that saccharification is carried out in the presence of urea by adding a urea solution to grains and starch, and then adding a saccharifying enzyme solution (Kumagaya, Arita, Akita)
Saccharification of raw starch in urea” (Presentation at the Japan Starch Society Conference).

The present invention is a development of this, and includes sodium hydroxide,
Alkalies such as potassium hydroxide have the ability to oxidize grains in relatively dilute solutions, eg, at a concentration of about 1.0%.

Regarding the alkali concentration during oxidation, it ranges from 0.5% to 1.
, 0% is particularly effective; if it exceeds 2%, the atomized liquid becomes extremely viscous and cannot be stirred sufficiently.

Furthermore, the sulfur components of proteins in grains are decomposed and cause unpleasant odors (eg, hydrogen sulfide and SH compounds).

Further, if the amount is less than 0.3%, it is difficult to obtain a sufficient effect of converting into alkali, and the risk of microbial contamination increases during busy times.

It was discovered that these drugs can be easily removed by neutralization or ion-exchange resin, and that saccharifying enzymes immediately act on them.
We discovered advantages different from urea.

According to the method of the present invention, grains can be heated to zero without being heated and steamed.
．． Since it is saccharified and saccharified in an alkaline solution of about 5 to 1.0%, the preparation and saccharification operations are easy, and it is a big advantage in that it does not require steaming and cooling equipment and the energy involved. The advantage is a) that it can be a powerful means of converting biomass into energy.

In addition, alcoholic fermented moromi can be used as a raw material for grain vinegars such as rice vinegar, can be used as a seasoning after being squeezed and filtered, and can be distilled to create alcoholic beverages that take advantage of the characteristics of each grain. It is also possible to manufacture products such as

In addition, it can be used to produce various substances obtained by converting glucose produced by saccharification.

In the present invention, glucoamylase activity was measured by the following method.

That is, using 1.0 ml of 2% soluble starch as a substrate,
Add 0.2 ml of 0.2 M acetate buffer (PH5,0) and 0.1 ml of enzyme solution, react at 40°C for 20 minutes, and
The reaction is stopped by adding 0.1 rul of N sodium hydroxide solution.

Neutralize with 0.1-2N hydrochloric acid, oxidize the glucose produced in the reaction solution with glucose oxidase, and oxidatively couple the generated hydrogen peroxide with 4-aminoantipyrine using peroxidase.
The measurement was carried out by the so-called glucostat method, in which a quinonemine dye with maximum absorption is produced and the colorimetric determination is performed (reaction at 40° C. for 20 minutes).

One unit of glucoamylase was defined as the activity of producing 1 μg of glucose in 60 minutes at 40°C.

Examples will be explained below.

Example 1 A method in which corn grits are saccharified in a sodium hydroxide solution, neutralized, and then saccharified and fermented using enzymes 1.0% sodium hydroxide solution 307 is added to 10 kg of corn grits (manufactured by Sunny Maize, AI), and the mixture is heated to room temperature while stirring occasionally. Soak overnight.

During this time, the corn grits will swell and coagulate, so 10%
Neutralize by adding 2.5 liters of hydrochloric acid.

Next, glucoamylase (Amano Pharmaceutical “Gluco 100J”)
) was added at 15 to 30 units per ml of saccharified solution, and heated at 35°C.
The mixture was allowed to act for 24 hours for saccharification.

After saccharification, reducing sugar was measured and found to be 652 fi/w
et kg, the saccharification rate was 87.8%.

Further, when the produced sugar was developed by paper chromatography using a developing solvent (n-butanol:pyridine:water-6:4:2), only glucose was detected as the main component.

Subsequently, yeast (5acch, cerevisia
e) was inoculated and fermented at 30°C for 2 days to obtain a fermented liquid with an alcohol concentration of 9.6%.

The corn grits used had a moisture content of 12.8% and a total sugar content of 7.
It was 43P/wetkg.

Example 2 Saccharification and fermentation method using corn as a raw material 10 kg of corn was coarsely crushed, 1.0% sodium hydroxide solution 301 was added thereto, and the corn was soaked overnight while being stirred occasionally to swell the corn.

Neutralize by adding 2.5 g of 10% hydrochloric acid, and add fusuma koji (fusuma koji) from filamentous fungi (Asp, oryzae) to this.
is the glucoamylase titer, which is 20 per 11 starch.
Add in units.

In this case, bran koji may be mixed with a culture solution of other saccharifying enzyme-producing bacteria, such as Asp, niger, Endomyces.
Mucor rouxi i 1 Rhizop
It can be replaced with US delemar etc.

At the same time as these saccharifying enzymes, cultured yeast (5acch, c
erevisiae), the number of bacteria is 108 per ml of culture solution.
and fermented at 30°C for 2 days.

The alcohol concentration of the fermented mash is 9.6%.
The fermentation rate was 85.5%.

Example 3 Method for saccharifying and fermenting white rice 10 kg of white rice (Nipponbare 90% white rice) was put into 1.0% sodium hydroxide solution 307, and left overnight with occasional stirring to swell and ferment the rice grains. .

Next, 2.5 kg of 10% hydrochloric acid solution was added to neutralize, and 2 kg of rice koji (Asp, oryzae) and 5 kg of cultured yeast (5 acch, cerevisiae) were added to this. (approximately 108/TILl) and 20°
Saccharification and fermentation is carried out by maintaining the temperature at a temperature above C and below 30°C.

Fermentation was completed in 2 days and an alcohol concentration of 10.2% was obtained.

This was distilled to obtain shochu stock solution 101 with an alcohol content of 29.3% and a distillation ratio of 88%.

In addition, for the purpose of producing seasoning liquid, an enzyme agent containing a strong protease (for example, "Clarin SJ" derived from Kikoji (yellow koji) fungus) is added at 20% per kilogram of white rice. When used in combination with ~5g, the production of amino acids increases.

Furthermore, to promote self-digestion of yeast in moromi,
Even after the fermentation was completed, a good seasoning liquid was obtained by keeping it at 30°C for 7 days.

The results of the analysis are as follows. Example 4 Saccharification and fermentation method using corn flour as raw material 10 kg of corn flour was put into 1.0% sodium hydroxide solution 301, and left overnight with occasional stirring.
Nako cornflour.

Next, 2.5% of a 10% hydrochloric acid solution was added to neutralize, and a commercially available saccharifying enzyme agent (for example, "Gluta 100J") was added to the raw material I.
50 units per Z and cultured yeast (5 ach.

cerevisiae) per culture ITL108
The mixture is then kept at 30°C for saccharification and fermentation.

Fermentation was completed in 4 days, and a fermented liquid with an alcohol concentration of 11.0% was obtained.

Example 5 Production of grain vinegar using white rice as a raw material 10 kg of white rice ("Nihonbare" 90% white rice) was put into a 0.5% sodium hydroxide solution 301 to swell and coagulate the rice grains.

Next, 2.5% of a 5% hydrochloric acid solution was added to neutralize it, an enzyme agent and yeast were added thereto, and fermentation was performed at 30°C for 4 days.
An alcohol concentration of 10.6%, an amino acid content of 6.7 and a direct reducing sugar content of 1.68% were obtained.

This was diluted twice with water, and acetic acid bacteria (Acetobacter) were added.
eraceti) and cultured at 30° C. for 15 days to obtain rice vinegar with an acetic acid concentration of 5.0%, an amino acid content of 3.1, and a direct reducing sugar of 0.75%.

Claims (1)

[Claims] Powdered or granular grains or crushed grains thereof are added by immersion into an alkaline solution with a concentration of 12% or less to form a paste, and after neutralization, enzymes or enzyme preparations are prepared. A saccharification and fermentation method for non-cooked grains, which is characterized by saccharification and fermentation using microorganisms. 2. The method for saccharification and fermentation of non-cooked grains according to claim 1, wherein the concentration of the alkaline solution is 1.0% or less and 0.5% or more.