JP3754850B2 - Immobilization carrier, method for producing fermentation product, and fermentation product - Google Patents

Description

【００５３】
〈発酵試験〉
実施例１，２及び比較例１で調製したアセチル化キトサン担体（固定化用担体）に酵母を担持させて固定化したもの（固定化微生物）を、図３に示す流動層型リアクター１（全容積：８０リットル）の反応タンク２内に設置し、以下の諸条件の下、前述の繰り返し回分発酵形式（反復回分式）によって主発酵を行い、後述する流動層形成率の測定、発酵性の評価、及び成分量の測定を実施した。尚、固定化用担体の下記所要量は、各実施例及び比較例の操作を繰り返し行って得た。
【００５４】
［主発酵条件］
１）担体：実施例１，２及び比較例１で調製したアセチル化キトサン担体
２）スケール：担体２８Ｌ、麦汁量４０Ｌ
３）使用酵母：ビール酵母（サッカロミセス・セレビシェ）
４）酵母固定化方法：常法に従い循環しながら２日間固定化用担体と接触させて固定化（担体２８Ｌに対して泥状酵母３ｋｇ）
５）液空塔速度：３ｃｍ／ｍｉｎ
６）発酵温度：８℃一定
７）発酵時間：４８時間／回
８）継続発酵回数：７回（回分発酵）
９）使用麦汁：糖度１１．５％Ｐｌａｔｏに調整した麦汁
【００５５】
（１）流動層形成率測定
従来の固定化用担体を用いた発酵においては、発酵で生成したガス成分が微生物等を担持した固定化用担体の内部に滞留し、固定化微生物の一部が流動層型リアクター１の反応タンク２内の原料液に浮上してしまい、固定化微生物が浮上層と流動層（浮上しない固定化用担体の層）に二分されてしまう。そこで、上記主発酵工程において、それら浮上層及び流動層の高さを測定し、下記式（９）；
η＝Ｈｒ／（Ｈｒ＋Ｈｆ）×１００…（９）
［式中、ηは流動層形成率（％）を示し、Ｈｒは流動層高さ（ｍｍ）を示し、Ｈｆは浮上層高さ（ｍｍ）を示す。］
の関係を用いて流動層形成率を求めた。その結果を表２に示す。比較例１で調製した固定化用担体（従来品）を用いた場合の流動層形成率が４６％であったのに対し、実施例１及び２で調製した本発明の固定化用担体を用いた場合の流動層形成率は、それぞれ１００％及び７１％と極めて高い値を示した。特に、実施例１の固定化用担体を用いれば浮上層が全く形成されず、このことより、本発明の優位性が確認された。尚、表２には、各固定化用担体に固定化された酵母数を測定した結果を併せて示す。この結果より、本発明の固定化用担体は、従来以上に酵母を固定化できることが確認された。
【００５６】
【表２】

【００５７】
（２）発酵性評価
発酵速度比較の指標として発酵開始２４時間のエキス消費量を振動式密度計（アントンパール社製、ＤＭＡ５８）により測定した。各回の測定値の平均値を上記表２に併せて示す。この値からは、本発明による固定化用担体を用いた場合及び従来品を用いた場合のその発酵速度（約６．５％／２４ｈｒ）に有意な差異は認められなかった。しかし、本発明の固定化用担体（実施例１及び２）を用いた場合には、各回の発酵速度は極めて安定して推移したのに対し、従来の固定化用担体（比較例１）を用いた場合には、発酵速度の増減が生じた。また、このとき、流動層リアクターには圧力負荷が発生し、実施例１及び２に比べて発酵が不安定なものであった。このことから、本発明の固定化用担体を用いれば、高い発酵効率を安定に維持でき、良好な発酵を実施できることが確認された。
【００５８】
（３）成分量測定
主発酵終了後、発酵終了液中の原麦汁エキス、真性エキス、及び発酵液中のアルコールを自動ビール分析装置（SERVOCHEM社製）により測定した。また、ジアセチル（ＤＡ）及び酢酸エチルの生成量をガスクロマトグラフィー（島津製作所社製、ＧＣ−１４Ｂ）により測定した。得られた結果を上記表２に併せて示す。本発明の固定化用担体（実施例１及び２）を用いた場合には、従来の固定化用担体（比較例１）を用いた場合よりも酢酸エチル量が多く、その他の成分はほぼ同等であった。
【００５９】
【発明の効果】
以上説明したように、本発明の固定化用担体によれば、固定化用担体に担持された微生物等による原料液の発酵によって生じるガス成分が内部に滞留することが防止され、流動層型リアクター内で良好な流動性を達成することができ、これにより発酵効率を高く維持しつつ安定な発酵を実施することができる。また、本発明の発酵生産物の製造方法によれば、そのような固定化用担体を用いることにより、発酵を高効率かつ安定的に行うことが可能となる。更に、本発明によれば、発酵が良好に行われるので、良好な香味を有する発酵生産物を得ることが可能となる。
【図面の簡単な説明】
【図１】本発明による固定化用担体の一実施形態の内部構造を模式的に示す断面図である。
【図２】本発明による固定化用担体の一実施形態の表面の一部を示す模式拡大図である。
【図３】本発明による発酵生産物の製造方法に好適な流動層型リアクターの一例を示す概略模式図である。
【図４】従来における固定化用担体の一例の内部構造を模式的に示す断面図である。
【符号の説明】
１…流動層型リアクター（バイオリアクター）、２…反応タンク、３…液循環部、４…整流部、５…流動層部、６…空筒部、７…ガス排出部、８…生産物タンク、９…原料液タンク、１０，２０…固定化用担体、１２，２２…細孔、１３…表面細孔（細孔）、１４…平滑面、２３…中空部。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an immobilization carrier, a method for producing a fermentation product, and a fermentation product. More specifically, the immobilization carrier capable of supporting microorganisms and / or enzymes, and the immobilization carrier are used. The present invention relates to a method for producing a fermented product, and a fermented product produced by the method for producing the fermented product.
[0002]
[Prior art]
In recent years, with the development of biotechnology, production of fermented products using bioreactors using immobilized microorganisms in the field of brewing such as malt alcoholic beverages (beer and happoshu using malt), wine, sake, vinegar, and soy sauce has been studied. By using a bioreactor in this way,
(1) Fermentation is carried out by immobilizing microorganisms and enzymes (hereinafter referred to as “microorganisms”) such as high concentrations of yeast, so that brewing can be completed quickly and the brewing period can be shortened. And construction costs can be reduced.
(2) Since continuous fermentation is possible, it is not necessary to input and collect microorganisms.
It is expected that.
[0003]
Examples of the bioreactor used for such fermentation include a complete mixing tank type, a packed bed type, a membrane type, a fluidized bed type, and a horizontal type reactor (reactor) because of the difference in the types. A fluidized bed reactor is suitable when fermentation using alcohol to produce alcohol and carbon dioxide gas, for example, main fermentation of beer or the like, by using a bioreactor to decompose brewing raw materials. In addition, the immobilized microorganism used in the bioreactor is one in which microorganisms or the like are supported and immobilized on a carrier, and the carrier used in the fluidized bed reactor does not inhibit the discharge of carbon dioxide gas or the like generated by fermentation. In addition, characteristics such as excellent wear resistance and fluidity are required.
[0004]
Examples of the carrier that satisfies such requirements include those made of various materials such as chitosan and sintered glass, but chitosan-based immobilization carriers are preferable from the viewpoint of wear resistance, fluidity, and the like. As such a chitosan-based immobilization carrier, a porous spherical body is generally used. FIG. 4 is a cross-sectional view schematically showing the internal structure of an example of such an immobilizing carrier. As shown in FIG. 4, the immobilization carrier 20 is provided with a plurality of pores 22 in a spherical body made of chitosan 21 and has a hollow portion 23 inside. And it arrange | positions in a bioreactor in the state in which microorganisms etc. were carry | supported by the inner surface of the pore 22 and the hollow part 23. FIG.
[0005]
[Problems to be solved by the invention]
When fermentation is performed using a fluidized bed reactor in which a microorganism or the like is supported on the immobilization carrier 20 having such a structure, part or most of the immobilization carrier 20 floats in the container. In some cases, the fluidized bed reactor sometimes becomes clogged. As a result, adverse effects such as inhibition of fluidization of the immobilization carrier 20, inhibition of discharge of gas components, and reduction in reaction efficiency (fermentation efficiency) may occur, and fermentation may become unstable.
[0006]
Therefore, the present invention has been made in view of such circumstances, the retention of gas components generated by fermentation is prevented, and good fluidity can be achieved in a fluidized bed reactor, thereby allowing fermentation. An object of the present invention is to provide an immobilization carrier capable of performing stable fermentation while maintaining high efficiency. Moreover, an object of this invention is to provide the manufacturing method of the fermented product which can perform fermentation highly efficiently and stably by using such a support | carrier for immobilization. Another object of the present invention is to obtain a fermented product that is well fermented and has a good flavor.
[0007]
[Means for Solving the Problems]
As a result of intensive studies to achieve the above object, the present inventors have found that the floating property of the carrier due to retention of gas components such as carbon dioxide inside the carrier for immobilization, and the internal structure and surface of the carrier for immobilization. We found that the structure is closely related. Based on this knowledge, further research has been conducted, and in the conventional immobilization support, there are many pores having extremely small diameters, and the gas components generated in the hollow portion by fermentation are not easily released to the outside. It was estimated. From this estimation, it was determined that the gas component stays in the hollow portion and buoyancy is imparted to the immobilization carrier, and the immobilization carrier floats. Thus, the immobilization carrier has been improved to arrive at the present invention.
[0008]
That is, the immobilization carrier of the present invention has a substantially spherical shape, and has pores on the surface and inside, and is disposed in the bioreactor in a state of supporting microorganisms and the like. From chitosan beads The average outer diameter is 1.5 to 4.0 mm, and the total area of the smooth surface when the surface is regarded as a smooth surface is 7.0 to 50.0 mm. ² And the pores open to the surface (hereinafter referred to as “surface pores”) are represented by the following formulas (1) and (2):
S1 ≦ S0 × 0.035 (1)
S2 ≧ S0 × 0.150 (2)
[Where S0 (mm ² ) Indicates the total area of the smooth surface, S1 (mm ² ) Has an opening area of 20 μm among the surface pores ² Indicates the total area of the following, S2 (mm ² ) Has an opening area of 300 μm among the surface pores ² Or more, preferably 300 to 3300 μm ² The total area of stuff is shown. ],
Preferably, the following formulas (5) and (6);
S1 ≦ S0 × 0.015 (5)
S2 ≧ S0 × 0.250 (6)
It is provided to satisfy the relationship shown in FIG.
[0009]
The immobilization support thus configured has a three-dimensional network structure as a whole by the surface pores and the pores provided inside (hereinafter referred to as “internal pores”). A sufficient amount of microorganisms or the like for carrying out a good fermentation of the liquid to be subjected to the above. Further, the immobilization carrier has a substantially spherical shape, and an average outer diameter of 1.5 to 4.0 mm. When the average outer diameter is less than 1.5 mm, it becomes difficult to carry a sufficient amount of microorganisms or the like on the immobilization carrier, and when the immobilization carrier is placed in a bioreactor, particularly a fluidized bed type reactor. In addition, it is difficult to ensure good fluidity of the immobilizing carrier, and the fermentation efficiency tends to decrease. On the other hand, when the average outer diameter exceeds 4.0 mm, the density of the carrier for immobilization in the bioreactor becomes sparse, the substantial contact area between the raw material liquid and the microorganisms decreases, and the fermentation efficiency decreases. There is a tendency. Further, since the immobilizing carrier has a spherical shape, the immobilizing carrier is less susceptible to damage and abrasion due to contact with each other and rubbing than when the immobilizing carrier has corners.
[0010]
When such a carrier for immobilization and the raw material liquid come into contact, the raw material liquid penetrates into the interior of the three-dimensional network structure, and the microorganisms and the like carried on the immobilization carrier are sufficiently in contact with the raw material liquid for fermentation. Done well. At this time, a gas component such as carbon dioxide gas is generated by fermentation of the raw material liquid by the microorganism or the like inside the immobilization carrier carrying microorganisms or the like, but the gas component is mostly retained inside the immobilization carrier. Without being released through the internal pores and surface pores of the immobilizing support.
[0011]
Here, when the surface of the immobilization carrier having the above average outer diameter is regarded as a smooth surface, that is, a substantially spherical surface, the total area of the smooth surface is 7.0 to 50.0 mm. ² It is said that. Of the surface pores, the opening area is 20 μm ² When the total area of the following (referred to as “micropores” for convenience) is 3.5% or less of the total area of the smooth surface as shown in the above formula (1), the immobilization carrier carrying microorganisms or the like It is effectively prevented that the gas component generated inside is blocked by such fine holes and is not easily released to the outside. Therefore, it is difficult for the gas component to stay in the immobilization carrier. Therefore, the immobilization carrier is prevented from floating on the raw material liquid, and good fluidity can be achieved. As a result, even when the immobilizing carrier is used in a fluidized bed reactor, stable fermentation can be performed while maintaining the fermentation efficiency of the raw material liquid extremely high as compared with the conventional case. Also, the opening area of the surface pores is 300 μm ² When the total area of the above (referred to as “large pores” for convenience) is 15% or more of the total area of the smooth surface as shown in the above formula (2), the gas generated inside the immobilizing carrier The components are more easily released to the outside. Therefore, it is possible to further prevent the gas component from staying inside the immobilization carrier, sufficiently prevent the immobilization carrier from floating on the raw material liquid, and reliably achieve good fluidity. Can do. Also, the area of such large pores is 3300 μm ² If it exceeds 1, it tends to be difficult to support a sufficient amount of microorganisms, etc., so the area of the large pores is 3300 μm. ² The following is desirable.
[0012]
Further, the immobilization carrier of the present invention has a substantially spherical shape, has pores on the surface and inside, and is disposed in the bioreactor in a state of supporting microorganisms. From chitosan beads The average outer diameter is 1.5 to 4.0 mm, and the total area of the smooth surface when the surface is regarded as a smooth surface is 7.0 to 50.0 mm. ² The number of pores opened on the surface per unit area of the smooth surface is 3000 to 9000 / mm. ² And the pores open to the surface are represented by the following formulas (3) and (4):
N1 ≦ N0 × 0.55 (3)
N2 ≧ N0 × 0.03 (4)
[In the formula, N0 (pieces) represents the total number of surface pores per unit area opened on the surface, and N1 represents an opening area of 20 μm among the surface pores. ² The total number per unit area of the following is shown, and N2 (pieces) has an opening area of 300 μm 2 or more, preferably 300 to 3300 μm among the surface pores. ² The total number of units per unit area is shown. Here, “unit area” indicates a unit area on the smooth surface. ],
Preferably, the following formulas (7) and (8);
N1 ≦ N0 × 0.45 (7)
N2 ≧ N0 × 0.10 (8)
It is provided to satisfy the relationship shown in FIG.
[0013]
The immobilization carrier having such a configuration is excellent in the release of gas components generated inside the immobilization carrier carrying microorganisms and the like, as in the above-described immobilization carrier of the present invention. Thus, the above-described immobilization carrier can be prevented from floating in the raw material liquid and good fluidity can be achieved. When the total number of micropores is 55% or less of the total number of surface pores as shown in the above formula (3), gas components generated inside the carrier for immobilization are blocked by the micropores. It is effectively prevented from becoming difficult to be released to the outside. Further, when the total number of large pores is 3% or more of the total number of surface pores as shown in the above formula (4), the gas component generated inside the immobilizing carrier is released to the outside. It becomes easy. Therefore, it is possible to sufficiently prevent the immobilization carrier from floating on the raw material liquid, and to reliably achieve good fluidity. As a result, stable fermentation can be performed while maintaining high fermentation efficiency of the raw material liquid. In such a configuration, the use of chitosan beads as an immobilization carrier is a feature of the present invention as a preferred embodiment.
[0014]
In addition, the method for producing a fermentation product of the present invention is a method for producing a fermentation product by performing fermentation using a bioreactor in which an immobilized microorganism is disposed. A fluidized bed type reactor having a fluidized bed portion in which immobilized microorganisms are arranged is used as a bioreactor using a carrier in which microorganisms are supported and immobilized.
[0015]
In such a fermented product production method, since the immobilization carrier of the present invention is used, the immobilization carrier carrying a conventional microorganism in the fluidized bed part floats on the raw material liquid. Is prevented, and the fluidity of the immobilizing carrier in the fluidized bed is improved. Therefore, the clogging of the fluidized bed reactor can be suppressed, and the possibility that the fermentation efficiency is lowered and the discharge of gas components is hindered can be significantly reduced. Therefore, fermentation can be performed efficiently and stably. As a result, the productivity and mass productivity of the fermented product can also be improved. Furthermore, when a fluidized bed reactor is used, there is an advantage that gas components generated by fermentation can be easily discharged out of the system as compared with other types of bioreactors.
[0016]
Further, in such a method for producing a fermented product, it is preferable that the fermented product obtained by fermentation is taken out from the fluidized bed type reactor, and a new raw material liquid is supplied to the fluidized bed type reactor so that the fermentation is repeated. It is.
[0017]
In addition, the method for producing a fermentation product of the present invention is a method for producing a fermentation product by performing fermentation using a bioreactor in which an immobilized microorganism is disposed. As a bioreactor, a fluidized bed part in which immobilized microorganisms are arranged and a fermentation liquid (fermentation progresses from the downstream side of the fluidized bed part). A fluidized bed reactor having a liquid circulating section that extracts a part of the raw material liquid) and returns it to the upstream side of the fluidized bed section.
[0018]
In such a fermented product production method, since the immobilization carrier of the present invention is used, a conventional immobilization carrier carrying microorganisms or the like floats in the raw material liquid in the fluidized bed portion. In addition, the fluidity of the immobilizing carrier in the fluidized bed is improved. Therefore, the clogging of the fluidized bed reactor can be suppressed, and the possibility that the fermentation efficiency is lowered and the discharge of gas components is hindered can be significantly reduced. Therefore, fermentation can be performed efficiently and stably. As a result, the productivity and mass productivity of the fermented product can also be improved. Furthermore, since the gas component generated during the fermentation is further prevented from staying in the fluidized bed due to the circulation of the fermentation broth or the like, the fermentation efficiency can be more reliably maintained. Furthermore, when a fluidized bed reactor is used, there is an advantage that gas components generated by fermentation can be easily discharged out of the system as compared with other types of bioreactors.
[0019]
Furthermore, fermentation is performed while forming a fluidized bed by extracting a part of the fermentation liquor from the downstream side of the fluidized bed part and returning it to the upstream side of the fluidized bed part, and the obtained fermentation product is removed from the fluidized bed type reactor. It is preferable to repeat the fermentation by taking out and supplying a new raw material liquid to the fluidized bed reactor. In this way, after the fermentation is finished, the circulation of the fermented liquid is stopped and the liquid after the fermentation is drawn out of the fluidized bed reactor, or immediately after that, a new raw material liquid is added to the fluidized bed reactor. Since it supplies, it becomes easy to perform fermentation by repeating and batching. In this way, the contact between the immobilized microorganisms and the raw material liquid is increased, the microorganisms during fermentation are propagated and the immobilized microorganisms are moderately renewed, and the distribution and activity of the microorganisms in the reactor are improved. Kept.
[0020]
In addition, since it is possible to perform a repeated batch process, the physiological state and growth behavior of microorganisms (fungal bodies) are similar to those of traditional liquor production, thereby making the fermented product product a better flavor. It tends to be granted. Therefore, according to the present invention, in addition to the merit of the production efficiency by using the conventional bioreactor that shortens the brewing period and does not require the input / recovery operation of microorganisms etc., it can be expected to provide a product that is preferable in terms of quality.
[0021]
Furthermore, examples of the microorganisms include yeasts, for example, liquor yeasts. When alcoholic yeasts are used, alcoholic beverages as fermentation products can be efficiently produced. Moreover, when brewer's yeast is used as liquor yeast, it becomes possible to produce a malt alcoholic beverage as liquor so as to exhibit an efficient and good flavor. In particular, the immobilization carrier of the present invention is particularly excellent in the ability to support yeasts such as liquor yeast and the release of gas components produced by fermentation as described above, so malt alcohol such as beer and sparkling liquor. When used for fermentation to produce beverages, the effect is maximized. And the fermented product of this invention is manufactured by the manufacturing method of the fermented product of this invention, It is characterized by the above-mentioned. The malt alcoholic beverage obtained by the method for producing a fermented product of the present invention has a very rich and gorgeous ester flavor.
[0022]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, preferred embodiments of the present invention will be described in detail. FIG. 1 is a cross-sectional view schematically showing the internal structure of an embodiment of an immobilization carrier according to the present invention. As shown in FIG. 1, the immobilization carrier 10 has a substantially spherical shape, and is provided with pores 12 on the surface and inside. For convenience of explanation, FIG. 1 shows the shape and quantity of the pores 12 different from the actual ones. As shown in FIG. 1, the immobilization carrier 10 does not have a large hollow portion inside, and a so-called three-dimensional network structure is formed by surface pores and internal pores. It is possible to carry microorganisms or the like on the inner surface of the glass. Then, the bioreactor is filled in a state where microorganisms or the like are supported on the inner surface of the pores 12 and fixed. Further, the average outer diameter of the immobilizing carrier 10 is 1.5 to 4.0 mm, and when the surface is regarded as a smooth surface, the smooth surface is substantially spherical, and the total area of the smooth surface is 7 .0 to 50.0mm ² It has become.
[0023]
FIG. 2 is a schematic enlarged view showing a part of the surface of the immobilizing carrier 10. As shown in FIG. 2, the immobilizing carrier 10 has an appearance in which a large number of surface pores 13 are provided on the smooth surface 14. Here, in the carrier 10 for immobilization, the surface pores 13 have the following formulas (1) to (4);
S1 ≦ S0 × 0.035 (1)
S2 ≧ S0 × 0.150 (2)
N1 ≦ N0 × 0.55 (3)
N2 ≧ N0 × 0.03 (4)
[Where S0 (mm ² ) Indicates the entire area of the smooth surface 14 which is substantially spherical, and S1 (mm ² ) Has an opening area of 20 μm in the surface pores 13. ² Indicates the total area of the following, S2 (mm ² ) Has an opening area of 300 μm among the surface pores 13. ² Or more, preferably 300 to 3300 μm ² N0 (number) indicates the total number per unit area of the surface pores 13 opened on the surface, and N1 indicates the opening area of the surface pores 13 is 20 μm. ² The total number per unit area of the following is shown, and N2 (pieces) has an opening area of 300 μm among the surface pores 13. ² Or more, preferably 300 to 3300 μm ² The total number of units per unit area is shown. The smooth surface 14 includes the opening surface of the surface pore 13. ],
Preferably the following formulas (5) to (8);
S1 ≦ S0 × 0.015 (5)
S2 ≧ S0 × 0.250 (6)
N1 ≦ N0 × 0.45 (7)
N2 ≧ N0 × 0.10 (8)
[Wherein S0, S1, S2, N0, N1 and N2 are the same as those in the above formulas (1) to (4). ]
It is provided so as to satisfy the relationship shown in FIG.
[0024]
The material of the immobilization carrier 10 is not particularly limited, and various known materials can be used, but chitin / chitosan, alginic acid, carrageenan, and cellulose are particularly preferable, and chitin / chitosan is particularly preferable. Chitin here is an N-acetylglucosamine homopolymer or a copolymer of N-acetylglucosamine and glucosamine that does not dissolve in an acid aqueous solution, and chitosan is a glucosamine homopolymer or glucosamine and N-acetylglucosamine. Of the copolymer, it is soluble in an acid aqueous solution. Chitosan-based beads preferable as the immobilization carrier 10 can be produced as described below, but chitosan to be used is not particularly limited, and the degree of acetylation, molecular weight, and the like may be appropriately selected. In general, sterilization is performed prior to immobilizing microorganisms or the like on such an immobilization carrier 10, but the sterilization method is arbitrary. For example, autoclaving, sterilization with caustic soda, etc. Method, steam sterilization method and the like are used, and among these, the pressure sterilization method or steam sterilization method is preferable. Further, the method for immobilizing microorganisms or the like on the immobilization carrier 10 is not particularly limited, and examples thereof include a method of adding the immobilization carrier 10 to a suspension of microorganisms and stirring or circulating the liquid, or Other known methods may be used.
[0025]
A method for producing the immobilization carrier 10 is not particularly limited, and for example, the following method can be used.
(1) First, chitosan having an average molecular weight of about tens of thousands and polyethylene glycol having an average molecular weight of about tens of thousands are dissolved in an acetic acid aqueous solution. After this solution is dropped into a mixed solution consisting of caustic soda, ethanol and water by a predetermined amount from a nozzle having a pore diameter of 0.1 to 0.4 mm, preferably 0.2 to 0.3 mm, and solidified and regenerated. Then, it is sufficiently washed with water until it becomes neutral to obtain a porous chitosan granule having an average outer diameter of 1.5 to 4.0 mm. The temperature of the mixed solution is preferably 15 to 40 ° C, more preferably 20 to 35 ℃. At this time, by adjusting the solution temperature or the like to adjust the deposition (coagulation) rate of chitosan, it is possible to obtain a favorable pore distribution characteristic of the immobilization carrier of the present invention. Since the size of the pores opened on the surface of the porous chitosan granule obtained here is smaller than that of the carrier of the present invention, it is preferable to subsequently carry out acid treatment.
[0026]
(2) This acid treatment is preferably performed at a temperature of 15 to 40 ° C., more preferably in a few percent aqueous acetic acid solution after removing the adhering water from the porous chitosan granules obtained as described in (1) above. What is necessary is just to wash with water until it becomes neutral immediately after immersion treatment for about several minutes at 20-35 degreeC. Thereby, the carrier for immobilization of the present invention can be obtained. When this carrier is subjected to fermentation, the carrier may be dissolved if the fermentation broth becomes acidic. Also good.
[0027]
(3) The acetylation treatment is performed by sufficiently replacing the water contained in the acid-treated porous chitosan granular material obtained in (2) above with ethanol, and then adding ethanol containing acetic anhydride, preferably 15 to 40 ° C, More preferably, the reaction is performed at a temperature of 20 to 35 ° C. for more than 10 hours. Thereafter, unreacted acetic anhydride is sufficiently removed with ethanol, and then the contained ethanol is sufficiently replaced with water.
[0028]
Examples of microorganisms to be supported on the immobilization carrier 10 include raw material liquids to be fermented and those suitable for the fermentation, such as yeast, lactic acid bacteria, acetic acid bacteria, and the like, which should be selected according to the purpose. Can do. Moreover, as Saccharomyces cerevisiae, Saccharomyces ubalum etc. can be used as yeast. Examples of the yeast form include aggregating and non-aggregating types, and any of them may be used.
[0029]
Here, for example, even in the same Saccharomyces cerevisiae, the cohesiveness of yeast is affected by brewing water, raw material composition, aeration conditions, yeast culture conditions, fermentation vessels, yeast handling, etc. The cohesive strength may change depending on the season. Thus, the yeast strains also have strong and weak cohesiveness, and even the same kind of yeast can be non-aggregating and cohesive depending on the strain. Which yeast is used can be appropriately selected depending on the fermentation product to be produced, the bioreactor, the fermentation conditions, and the like. For example, when it is necessary to prevent sedimentation or accumulation in the bioreactor vessel and maintain the number of floating yeasts and fermentation efficiency in the raw material liquid, it may be desirable to use non-aggregating ones. is there. In addition, when an enzyme is supported on an immobilization carrier, these enzymes can include enzymes produced by the above-mentioned microorganisms, and these enzymes can be immobilized on an immobilization carrier by a known method. Good.
[0030]
According to the immobilization carrier 10 configured in this way, the whole surface has a three-dimensional network structure by the surface pores 13 and the internal pores, and a sufficient amount of microorganisms to perform good fermentation of the raw material liquid Etc. can be carried. Therefore, the fermentation efficiency of the raw material liquid can be sufficiently increased. In addition, the immobilization carrier 10 has a substantially spherical shape, and if the average outer diameter is less than 1.5 mm, it becomes difficult to carry a sufficient amount of microorganisms or the like on the immobilization carrier 10 and immobilization is performed. When the carrier 10 is placed in a bioreactor, particularly a fluidized bed reactor, it is difficult to ensure good fluidity and the fermentation efficiency tends to decrease. On the other hand, when the average outer diameter exceeds 4.0 mm, the density of the carrier 10 for immobilization in the bioreactor becomes sparse, the substantial contact area between the raw material liquid and microorganisms, etc. decreases and the fermentation efficiency decreases. Tend to. Therefore, since the average outer diameter of the immobilizing carrier 10 of the present invention is set to a value within the above range, the fermentation efficiency can be stably maintained at a high level.
[0031]
Furthermore, the immobilization carrier 10 is substantially spherical and is less susceptible to damage and wear due to mutual contact and abrasion of the immobilization carrier 10 than when the immobilization carrier 10 has corners. Therefore, the immobilization carrier 10 is scraped off during use (during flow), and the carried microorganisms and the like are sufficiently prevented from dropping off, so that the fermentation efficiency is reduced and the bioreactor container is blocked. It can be effectively prevented.
[0032]
Further, when the immobilization carrier 10 and the raw material liquid come into contact, the raw material liquid penetrates into the interior of the three-dimensional network structure, and the microorganisms and the like carried on the immobilization carrier 10 are sufficiently in contact with the raw material liquid for fermentation. Is done well. At this time, a gas component such as carbon dioxide gas is generated inside the immobilization carrier 10, but the gas component hardly stays inside the immobilization carrier 10, and the internal pores of the immobilization carrier 10. And discharged through the surface pores 13 to the outside. That is, when the surface of the immobilizing carrier 10 is regarded as a substantially spherical smooth surface 14, the opening area of the surface pores 13 is 20 μm. ² When the total area of the following (micropores) is 3.5% or less of the total area of the smooth surface 14 as shown in the above formula (1), the gas components generated inside the immobilizing carrier 10 are It is sufficiently prevented from being blocked by such fine holes and becoming difficult to be released to the outside.
[0033]
Further, when the total area of these micropores satisfies the above formula (5), it is possible to prevent the gas component from being blocked more sufficiently. Therefore, it is difficult for the gas component to stay inside the immobilization carrier 10, and the immobilization carrier 10 is prevented from floating on the raw material liquid, and good fluidity can be achieved. Therefore, when the immobilization carrier 10 is used in a fluidized bed reactor, the fermentation efficiency of the raw material liquid can be maintained extremely high and stable as compared with the conventional case.
[0034]
Further, the area of the surface pores 13 is 300 μm. ² When the total area of the above (large pores) is 15% or more of the total area of the smooth surface 14 as shown in the above formula (2), the gas component generated inside the immobilizing carrier 10 is extremely high. Easily released to the outside. When the total area of these large pores satisfies the above formula (6), the gas component is more easily released to the outside. As a result, it is possible to further prevent the gas component from staying inside the immobilization carrier 10, and the floating in the raw material liquid of the immobilization carrier 10 is further suppressed and the fluidity is improved. It is possible to carry out stable fermentation while maintaining the fermentation efficiency extremely high. Moreover, the area of the large pore is 3300 μm ² If it exceeds the range, it tends to be difficult to carry a sufficient amount of microorganisms.
[0035]
Furthermore, the total number of fine pores per unit area is 55% or less, preferably 45% or less of the total number of surface pores 13 as shown in the above formula (3), preferably the above formula (7). Therefore, blocking of gas components by such fine holes is further prevented. On the other hand, the total number of large pores per unit area is 3% or more, preferably 10% or more of the total number of surface pores 13 as shown in the above formula (4), preferably the above formula (8). The gas component generated inside the immobilizing carrier 10 is very easily released to the outside. Therefore, the immobilization carrier 10 is more reliably prevented from floating on the raw material liquid, and the fluidity of the immobilization carrier 10 in the raw material liquid can be further improved. Therefore, the fermentation efficiency of the raw material can be maintained better and more stably.
[0036]
Further, when the immobilization carrier 10 is made of chitosan, for example, formed of chitosan-based beads, the chitosan-based substance is hydrophilic and rich in porosity, so that the discharge of gas components becomes extremely easy. Since it is hard to be worn away and its density is close to the specific gravity of a commonly used raw material liquid, the fluidity in the raw material liquid is further improved. Furthermore, since chitosan-based beads have a relatively large amount of microorganisms or the like that can be held, the fermentation time of the raw material liquid tends to be shortened, and the microorganisms and the like are immobilized relatively slowly. This facilitates the growth and removal of microorganisms.
[0037]
Next, the manufacturing method of the fermented product of this invention is demonstrated. First, in a bioreactor suitable for the method for producing a fermented product of the present invention, immobilized microorganisms that are immobilized by supporting microorganisms or the like on the above-described immobilization carrier 10 are disposed inside, and a raw material liquid is disposed therein. And the immobilized microorganisms are brought into contact with each other for fermentation. Examples of such bioreactors include complete mixing tank reactors, packed bed reactors, membrane reactors, fluidized bed reactors, horizontal reactors, etc. A fluidized bed reactor is preferred for fermentation in which alcohol and carbon dioxide are produced by decomposition of the raw material.
[0038]
In the method for producing a fermented product according to the present invention, as such a fluidized bed type reactor, a fluidized bed part in which immobilized microorganisms are arranged, and a fermentation liquid (fermentation proceeds from the downstream side of the fluidized bed part). And a liquid circulation part that extracts a part of the raw material liquid) and returns it to the upstream side of the fluidized bed part. FIG. 3 is a schematic diagram showing an example of a fluidized bed reactor suitable for the method for producing a fermentation product according to the present invention. A fluidized bed reactor 1 shown in FIG. 3 includes a reaction tank 2 and a liquid circulation unit 3, and the reaction tank 2 includes a rectifying unit 4, a fluidized bed unit 5, and an empty cylinder unit 6 in order from the upstream side. A gas discharge part 7 is provided at the downstream end of the reaction tank 2. The liquid circulation unit 3 includes a pipe 31 connected to the downstream side of the fluidized bed unit 5 and the upstream side of the fluidized bed unit 5 (rectifying unit 4 in FIG. 3), a pump 32, and a valve 33. The pipe 31 is branched in the middle and connected to the product tank 8 via a valve 34. A raw material liquid tank 9 is connected to the upstream end of the reaction tank 2 by a pipe 92 having a pump 91.
[0039]
The fluidized bed part 5 is a part in which an immobilizing carrier 10 that is immobilized by supporting microorganisms or the like is disposed inside, and the liquid circulating part 3 is a part of the fermentation broth supplied to the reaction tank 2. This is a portion that is extracted from the downstream side of the fluidized bed portion 5 and returned to the upstream side of the fluidized bed portion 5 of the reaction tank 2 again. In the fluidized bed reactor 1 shown in FIG. 3, the extracted fermentation liquor is returned from the rectification unit 4 into the reaction tank 2 and the flow of the introduced liquid is adjusted here. Moreover, the empty cylinder part 6 is a part which gas-liquid-separates gas components, such as a carbon dioxide gas produced during fermentation, and fermentation liquid, and specifically, the part from the liquid level to the gas discharge part 7 of the upper part of the reaction tank 2 It is. And gas components, such as a carbon dioxide gas isolate | separated by the empty cylinder part 6, are discharged | emitted from the gas discharge part 7 out of a container.
[0040]
In the fluidized bed reactor 1 configured as described above, the immobilizing carrier 10 that supports and immobilizes microorganisms or the like is disposed in the fluidized bed portion 5, and the raw material liquid is supplied from the raw material liquid tank 9 using the pump 91. The rectification unit 4 is supplied to perform fermentation. Then, a part of the fermentation broth is extracted from the downstream side of the fluidized bed part 5 using the pump 32 of the liquid circulation part 3 and returned again to the upstream side of the fluidized bed part 5 of the reaction tank 2 (rectifying part 4 in FIG. 3). And fermenting while forming a fluidized bed. The carbon dioxide gas generated during the fermentation is discharged out of the container from the gas discharge part 7 through the empty cylinder part 6 without staying in the fluidized bed by circulating the fermentation liquid or the like. The liquid superficial velocity during fermentation (linear velocity of fluid per fluidized bed unit volume) can be varied depending on the density of the carrier 10 for immobilization, but is preferably 1 to 20 cm / min, preferably 1 to 12 cm / min. min is more preferable.
[0041]
According to the method for producing a fermented product using such a fluidized bed reactor 1, the immobilization carrier 10 according to the present invention is used. Is sufficiently prevented from floating, and the fluidity of the immobilizing carrier 10 in the fluidized bed portion 5 is improved. Therefore, the clogging of the container of the fluidized bed reactor 1 is suppressed, and the possibility that the fermentation efficiency is lowered and the discharge of gas components is hindered can be significantly reduced. As a result, fermentation can be performed efficiently and stably, and productivity and mass productivity of the fermentation product can be improved. Furthermore, since the gas component produced | generated during fermentation is prevented from staying in the fluidized bed part 5 by circulating a fermentation liquor etc., it becomes possible to maintain fermentation efficiency still higher. Furthermore, the use of the fluidized bed reactor 1 has the advantage that the gas components produced by fermentation can be easily discharged out of the system as compared with other types of bioreactors. Can be achieved reliably.
[0042]
Further, in the fluidized bed reactor 1 shown in FIG. 3, after the fermentation is completed, the fermentation finished liquid containing the fermentation product is taken out from the fluidized bed reactor 1 to the product tank 8 using the pump 32 and the valve 34. It is preferable to repeat the fermentation by supplying a new raw material liquid from the raw material liquid tank 9 to the fluidized bed reactor 1 using the pump 91. That is, after the fermentation is finished, the circulation is stopped and the fermentation finished liquid is taken out of the fluidized bed reactor 1 at the same time or immediately after that, by supplying a new raw material liquid to the fluidized bed reactor 1. The fermentation is repeated. As a method for operating a bioreactor such as the fluidized bed reactor 1, any of a batch type, a repeated batch type, and a continuous type is possible. By repeatedly performing fermentation as described above, a repeated batch type fermentation treatment is performed. Is extremely easy to implement. Therefore, a product having a good flavor can be obtained in a short time.
[0043]
In addition, microorganisms are grown and renewed during fermentation, a good distribution of microbial cells in the fluidized bed reactor 1 is achieved, and the physiological state and growth behavior of microorganisms (bacterial cells) are traditional liquors. Since it is similar to the batch-type operation which is the production method of the present invention, it is possible to improve the flavor of the product by carrying out the production method of the fermented product of the present invention by repeated batch-type.
[0044]
Moreover, as a raw material liquid for manufacturing a fermentation product, what is suitable for fermentation by microorganisms, such as a yeast to be used, should just be used, and a known thing can be used arbitrarily. For example, for the production of alcoholic beverages, usually, wort, fruit juice, sugar solution, cereal saccharified solution and the like can be used alone or mixed appropriately to form a raw material solution. In addition, it is also preferable to add an appropriate nutrient or the like to the raw material liquid as necessary. Furthermore, the fermentation conditions are basically the same as the known conditions. For example, in the case of malt alcoholic beverage brewing, the fermentation temperature is usually 15 ° C. or lower, preferably 8 to 10 ° C., and in the case of wine brewing. Usually 20 ° C. or lower, preferably 15 to 20 ° C.
[0045]
The fermented product that can be produced by the method for producing a fermented product of the present invention includes fermented products in various brewing fields such as malt alcoholic beverages, wine, sake, vinegar, and soy sauce. Alcoholic beverages such as wine and sake are preferred. Among these, the immobilization carrier of the present invention is excellent in the supportability of yeasts such as liquor yeasts, and is excellent in the release of gas components produced by fermentation as described above, so its effect From the viewpoint of drawing out the maximum amount, malt alcoholic beverages such as beer and happoshu are preferable. And the malt alcoholic beverage obtained by the manufacturing method of the fermented product of this invention has a very favorable flavor, for example, has a rich and gorgeous ester flavor.
[0046]
Moreover, since fermentation is performed stably and satisfactorily, the original flavor of the malt is not impaired, and conversely, the flavor is emphasized and a rich flavor is imparted. Further, grains other than malt may be added to the raw material, and in that case, the flavor and taste of these raw materials are not impaired, and each flavor can be tastefully harmonized individually. Therefore, according to the present invention, in addition to the merit of the production efficiency by using the conventional bioreactor that shortens the brewing period and does not require the input / recovery operation of microorganisms etc., it can be expected to provide a product that is preferable in terms of quality.
[0047]
【Example】
EXAMPLES Hereinafter, although the content of this invention is demonstrated more concretely by an Example and a comparative example, this invention is not limited to these Examples at all. “%” Indicating the content ratio indicates “wt%”.
[0048]
<Example 1>
80 g of chitosan having a degree of deacetylation of 78%, an average molecular weight of 52,000, and 100 g of polyethylene glycol having an average molecular weight of 20000 were dissolved in a 5% aqueous acetic acid solution so that the total amount became 1000 ml. This solution was dropped into a mixed solution consisting of 5% caustic soda, 20% ethanol, and 75% water at 25 ° C by a predetermined amount from a nozzle having a pore diameter of 0.25 mm to solidify and regenerate, and then washed thoroughly with water until it became neutral. Thus, 1 L of a porous chitosan granule having an average outer diameter of 3.5 mm (liter; the same applies hereinafter) was obtained. After removing the adhering water from the obtained porous chitosan granular material, it was immersed in 1 liter of 1% aqueous acetic acid at 25 ° C. for 1 minute, and then immediately washed with water until it became neutral, and an acid having an average outer diameter of 3.0 mm. 0.7 L of processed porous chitosan granules were obtained. The water contained in the obtained acid-treated porous chitosan granules was sufficiently substituted with ethanol, 0.7 L of ethanol containing 63 g of acetic anhydride was added, and the mixture was reacted at 25 ° C. for 16 hours. Unreacted acetic anhydride was sufficiently removed with ethanol, and then the contained ethanol was sufficiently replaced with water to obtain 0.7 L of an acetylated chitosan carrier (an immobilizing carrier) having an average outer diameter of 3.0 mm.
[0049]
<Example 2>
80 g of chitosan having a degree of deacetylation of 78%, an average molecular weight of 52,000, and 100 g of polyethylene glycol having an average molecular weight of 20000 were dissolved in a 5% aqueous acetic acid solution so that the total amount became 1000 ml. This solution was dropped into a mixed solution consisting of 5% caustic soda, 20% ethanol, and 75% water at 25 ° C by a predetermined amount from a nozzle having a pore diameter of 0.25 mm to solidify and regenerate, and then washed thoroughly with water until it became neutral. Thus, 1 L of porous chitosan granules having an average outer diameter of 3.5 mm were obtained. After removing the adhering water from the obtained porous chitosan granular material, it was immersed in 1 L of 1% acetic acid aqueous solution at 25 ° C. for 2 minutes, then immediately washed with water until neutral, and an acid having an average outer diameter of 3.0 mm. 0.6 L of treated porous chitosan granules were obtained. The water contained in the obtained acid-treated porous chitosan granules was sufficiently substituted with ethanol, and then 0.6 L of ethanol containing 54 g of acetic anhydride was added and reacted at 25 ° C. for 16 hours. Unreacted acetic anhydride was sufficiently removed with ethanol, and then the contained ethanol was sufficiently replaced with water to obtain 0.6 L of an acetylated chitosan carrier (an immobilizing carrier) having an average outer diameter of 3.0 mm.
[0050]
<Comparative example 1>
80 g of chitosan having a degree of deacetylation of 78%, an average molecular weight of 52,000, and 100 g of polyethylene glycol having an average molecular weight of 20000 were dissolved in a 4% aqueous acetic acid solution so that the total amount became 1000 ml. This solution was dropped into a mixed solution of 5% caustic soda, 20% ethanol, and 75% water at 35 ° C by a predetermined amount from a nozzle having a pore diameter of 0.25 mm to solidify and regenerate, and then washed thoroughly with water until neutral. Thus, 1 L of porous chitosan granules having an average outer diameter of 3.5 mm were obtained. After removing the adhering water from the obtained porous chitosan granule, it was immersed in 1 L of 0.5% acetic acid solution at 25 ° C. for 1 minute, and then immediately washed with water until it became neutral, with an average outer diameter of 3.0 mm. 0.8 L of acid-treated porous chitosan granules was obtained. The water contained in the obtained acid-treated porous chitosan granules was sufficiently substituted with ethanol, and then 0.8 L of ethanol containing 72 g of acetic anhydride was added and reacted at 25 ° C. for 16 hours. Unreacted acetic anhydride was sufficiently removed with ethanol, and then the contained ethanol was sufficiently replaced with water to obtain 0.8 L of an acetylated chitosan carrier (an immobilizing carrier) having an average outer diameter of 3.0 mm.
[0051]
<Surface analysis of immobilization carrier>
The average outer diameter of the acetylated chitosan carrier (immobilization carrier) prepared in Examples 1 and 2 and Comparative Example 1 was measured by measuring the outer diameter of the particles with an optical microscope equipped with a micrometer. The total area of the smooth surface when the surface of the carrier for immobilization was regarded as a smooth surface was calculated from the obtained average outer diameter. Further, the surface is magnified and observed with an electron microscope to obtain a magnified image, and the image is analyzed with an image processing apparatus (Image Analyzer V20, manufactured by Toyobo Co., Ltd.). The number of pores inside and the open area were measured. Based on the total area of the obtained smooth surface, the area of the observation field, and the opening area of the pores, the micropores (opening area is 20 μm) with respect to the total area of the smooth surface. ² The following pores) and large pores (opening area is 300 μm) ² The area ratio (opening ratio) on the smooth surface of the above pores) was calculated. Further, the distribution of the number of pores was determined based on the opening area of the pores and the count value. The results are summarized in Table 1.
[0052]
[Table 1]

[0053]
<Fermentation test>
A fluidized bed reactor 1 (all immobilized microorganisms) shown in FIG. 3 was obtained by immobilizing yeast on an acetylated chitosan carrier (an immobilizing carrier) prepared in Examples 1 and 2 and Comparative Example 1 (immobilized microorganism). (Volume: 80 liters) in the reaction tank 2 and under the following conditions, the main fermentation is carried out by the above-mentioned repeated batch fermentation method (repeated batch method). Evaluation and measurement of component amounts were carried out. The following required amount of the carrier for immobilization was obtained by repeating the operations of the examples and comparative examples.
[0054]
[Main fermentation conditions]
1) Carrier: Acetylated chitosan carrier prepared in Examples 1 and 2 and Comparative Example 1
2) Scale: carrier 28L, wort amount 40L
3) Yeast used: Beer yeast (Saccharomyces cerevisiae)
4) Yeast immobilization method: Immobilization by contacting with a carrier for immobilization for 2 days while circulating according to a conventional method (3 kg of muddy yeast with respect to the carrier 28L)
5) Liquid superficial velocity: 3 cm / min
6) Fermentation temperature: 8 ° C constant
7) Fermentation time: 48 hours / time
8) Number of continuous fermentation: 7 times (batch fermentation)
9) Wort used: wort adjusted to sugar level 11.5% Plato
[0055]
(1) Fluidized bed formation rate measurement
In fermentation using a conventional immobilization carrier, gas components generated by fermentation stay inside the immobilization carrier carrying microorganisms and the like, and a part of the immobilized microorganisms is a reaction tank of the fluidized bed reactor 1. 2 is floated on the raw material solution in 2 and the immobilized microorganisms are divided into a floating layer and a fluidized bed (a layer of an immobilizing carrier that does not float). Therefore, in the main fermentation step, the heights of the floating layer and the fluidized bed are measured, and the following formula (9);
η = Hr / (Hr + Hf) × 100 (9)
[In the formula, η represents a fluidized bed formation rate (%), Hr represents a fluidized bed height (mm), and Hf represents a floating layer height (mm). ]
The fluidized bed formation rate was determined using the above relationship. The results are shown in Table 2. When the immobilization carrier prepared in Comparative Example 1 (conventional product) was used, the fluidized bed formation rate was 46%, whereas the immobilization carrier of the present invention prepared in Examples 1 and 2 was used. The fluidized bed formation rate was 100% and 71%, respectively. In particular, when the immobilization support of Example 1 was used, no floating layer was formed, and this confirmed the superiority of the present invention. Table 2 also shows the results of measuring the number of yeasts immobilized on each immobilization carrier. From this result, it was confirmed that the immobilization carrier of the present invention can immobilize yeast more than before.
[0056]
[Table 2]

[0057]
(2) Fermentability evaluation
As an index for comparing fermentation rates, the amount of extract consumed for 24 hours from the start of fermentation was measured with a vibratory density meter (manufactured by Anton Paar, DMA58). The average value of each measurement value is also shown in Table 2 above. From this value, no significant difference was observed in the fermentation rate (about 6.5% / 24 hr) when using the immobilization carrier according to the present invention and when using the conventional product. However, when the immobilization carrier of the present invention (Examples 1 and 2) was used, the fermentation rate of each round was extremely stable, whereas the conventional immobilization carrier (Comparative Example 1) was changed. When used, the fermentation rate increased or decreased. At this time, a pressure load was generated in the fluidized bed reactor, and the fermentation was unstable as compared with Examples 1 and 2. From this, it was confirmed that if the immobilization carrier of the present invention is used, high fermentation efficiency can be stably maintained and good fermentation can be performed.
[0058]
(3) Component content measurement
After the main fermentation was completed, the raw wort extract, the true extract, and the alcohol in the fermentation broth were measured with an automatic beer analyzer (manufactured by SERVOCHEM). Moreover, the production amounts of diacetyl (DA) and ethyl acetate were measured by gas chromatography (manufactured by Shimadzu Corporation, GC-14B). The obtained results are also shown in Table 2 above. When the immobilization carrier of the present invention (Examples 1 and 2) was used, the amount of ethyl acetate was larger than when the conventional immobilization carrier (Comparative Example 1) was used, and the other components were almost the same. Met.
[0059]
【The invention's effect】
As described above, according to the immobilization carrier of the present invention, gas components generated by fermentation of the raw material liquid by microorganisms or the like supported on the immobilization carrier are prevented from staying inside, and the fluidized bed reactor In this way, good fluidity can be achieved, whereby stable fermentation can be carried out while maintaining high fermentation efficiency. Moreover, according to the method for producing a fermented product of the present invention, fermentation can be performed with high efficiency and stability by using such an immobilizing carrier. Furthermore, according to this invention, since fermentation is performed favorably, it becomes possible to obtain a fermented product having a good flavor.
[Brief description of the drawings]
FIG. 1 is a cross-sectional view schematically showing the internal structure of an embodiment of an immobilization carrier according to the present invention.
FIG. 2 is a schematic enlarged view showing a part of the surface of an embodiment of an immobilization carrier according to the present invention.
FIG. 3 is a schematic diagram showing an example of a fluidized bed reactor suitable for the method for producing a fermentation product according to the present invention.
FIG. 4 is a cross-sectional view schematically showing an internal structure of an example of a conventional immobilization carrier.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Fluidized bed reactor (bioreactor), 2 ... Reaction tank, 3 ... Liquid circulation part, 4 ... Rectification part, 5 ... Fluidized bed part, 6 ... Empty cylinder part, 7 ... Gas discharge part, 8 ... Product tank , 9 ... Raw material liquid tank, 10, 20 ... Immobilization carrier, 12, 22 ... Fine pore, 13 ... Surface fine pore (pore), 14 ... Smooth surface, 23 ... Hollow portion.

Claims (10)

A substantially spherical, and pores provided surface and inside, are arranged in the bioreactor while carrying microorganisms and / or enzymes, a carrier for immobilizing ing chitosan based beads,
The average outer diameter is 1.5 to 4.0 mm,
When the surface is regarded as a smooth surface, the total area of the smooth surface is 7.0 to 50.0 mm ² ;
A carrier for immobilization, wherein the pores open on the surface are provided so as to satisfy a relationship represented by the following formulas (1) and (2).
S1 ≦ S0 × 0.035 (1)
S2 ≧ S0 × 0.150 (2)
[In the formula, S0 (mm ² ) represents the total area of the smooth surface, and S1 (mm ² ) represents the total area of the pores having an opening area of 20 μm ² or less among the pores opening on the surface. S2 (mm ² ) indicates the total area of the pores having an opening area of 300 μm ² or more among the pores opened on the surface. ] A substantially spherical, and pores provided surface and inside, are arranged in the bioreactor while carrying microorganisms and / or enzymes, a carrier for immobilizing ing chitosan based beads,
The average outer diameter is 1.5 to 4.0 mm,
When the surface is regarded as a smooth surface, the total area of the smooth surface is 7.0 to 50.0 mm ² ;
The number of pores opened on the surface per unit area of the smooth surface is 3000 to 9000 / mm ² ;
A carrier for immobilization, wherein the pores opened on the surface are provided so as to satisfy a relationship represented by the following formulas (3) and (4).
N1 ≦ N0 × 0.55 (3)
N2 ≧ N0 × 0.03 (4)
[In the formula, N0 (pieces) represents the total number of the pores per unit area opened on the surface, and N1 is the total per unit area of the pores having an opening area of 20 μm ² or less. N2 (number) indicates the total number per unit area of the pores having an opening area of 300 μm ² or more among the pores opened on the surface. ] In a method for producing a fermentation product by performing fermentation using a bioreactor in which immobilized microorganisms are disposed,
As the immobilizing microorganism, an immobilizing carrier according to claim 1 or 2 that is supported by immobilizing a microorganism and / or an enzyme is used,
As the bioreactor, a fluidized bed type reactor having a fluidized bed portion in which the immobilized microorganism is disposed is used. The fermentation production according to claim 3 , wherein the fermentation product obtained by the fermentation is taken out from the fluidized bed reactor and a new raw material liquid is supplied to the fluidized bed reactor to repeatedly perform the fermentation. Manufacturing method. In a method for producing a fermentation product by performing fermentation using a bioreactor in which immobilized microorganisms are disposed,
As the immobilizing microorganism, an immobilizing carrier according to claim 1 or 2 that is supported by immobilizing a microorganism and / or an enzyme is used,
As the bioreactor, a fluidized bed part in which the immobilized microorganisms are disposed, and a liquid circulation part that extracts a part of the fermentation liquid from the downstream side of the fluidized bed part and returns it to the upstream side of the fluidized bed part. A method for producing a fermentation product, comprising using a fluidized bed reactor provided. Performing fermentation while forming a fluidized bed by extracting a part of the fermentation liquor from the downstream side of the fluidized bed part and returning it to the upstream side of the fluidized bed part,
Production of the resulting fermentation products a fermentation product according to claim 5, wherein the supplying a new material liquid to flow dynamic bed reactor repeating performing the fermentation is taken out from the fluidized bed reactor Method. The said microorganism is yeast, The manufacturing method of the fermented product as described in any one of Claims 3-6 characterized by the above-mentioned. The said yeast is liquor yeast, The said fermentation product is liquor, The manufacturing method of the fermentation product of Claim 7 characterized by the above-mentioned. The method for producing a fermented product according to claim 8, wherein the liquor yeast is beer yeast, and the liquor is a malt alcoholic beverage. A fermented product produced by the method for producing a fermented product according to any one of claims 3 to 9 .

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