WO1997013850A1

WO1997013850A1 - Living-yeast immobilization method and application device thereof

Info

Publication number: WO1997013850A1
Application number: PCT/CN1996/000083
Authority: WO
Inventors: Yaodong Wang
Original assignee: Yaodong Wang
Priority date: 1995-10-06
Filing date: 1996-10-07
Publication date: 1997-04-17
Also published as: AU7209696A; CN1147553A; CN1063487C

Abstract

The present invention relates to a living-yeast immobilization method and to an application device thereof. During the living-yeast free fermentation, there is a need to exchange seed frequently, and it is easy to be contaminated by the other bacterium. To overcome these shortcomings and to increase fermentative efficiency, the present invention provides an immobilization method, wherein the living-yeast inoculum is mixed with soluble alginate solution, the sponge is dipped into the mixture, the sponge filled with the mixture is taken out and placed into 2-10 % (w/w) soluble calcium salt solution. Once the calcium alginate gel has formed and filled the sponge cavity, the immobilized living-yeast carrier is formed. An application device specific for said immobilized living-yeast carrier fermentation is also provided. The carrier life, fermentative efficiency and economic benefit are all increased in the invention.

Description

Technical field

The invention relates to a method for fixing live yeast and a device used in the fermentation technology.

Background technique

In the specialized brewing industry, such as the use of free yeast abortion in the process of producing alcohol, the yeast also flows with the sap flowing, so that it is most susceptible to infectious bacteria. Generally, it must be changed once a week to maintain normal production. Production is inconvenient, and production has to be stopped when changing seeds, which also brings economic losses. In addition, when there is a bacterial infection, the fermentation rate will be reduced, the mature mash liquid will contain less alcohol, the residual sugar will increase, the fermentation by-products will increase, the production cost will increase, and the economic benefits will be reduced. Therefore, people are scrambling to find a method that is conducive to the proliferation of live yeast to reduce contamination by bacteria. The so-called method of immobilizing live yeast is to highly concentrate the live yeast on the carrier and continue to grow and proliferate to form a high concentration of bioenzyme catalyst, thereby greatly accelerating the reaction speed and making the bioreactor viable? ^ Ability has been greatly improved, and greater economic benefits have been achieved than traditional technology. The technology of producing ethanol from immobilized proliferative yeast carrier is a highly sophisticated technology in the field of contemporary bioengineering. Therefore, many foreign countries are also competing for research, but the progress is in the laboratory stage.

At present, the method for fixing live yeast uses calcium alginate gel to embed the live yeast in it, and it becomes a spherical or film carrier, and some are added with a plastic carrier enhancer; for example, Chinese patent S 5 1 0 5: 7 9 uses this carrier to fix live yeast. The disadvantages are poor resistance to erosion and abrasion, and it is difficult to change the shape and size of the carrier at any time according to different process needs.

Disclosure of invention

The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art, to overcome the shortcomings of frequent replacement of free yeast in the fermentation process, and to prevent the infection of miscellaneous bacteria during the fermentation process, improve the fermentation efficiency and strengthen the live yeast. The fixing in the carrier makes the carrier of the fixed live yeast resistant to erosion and abrasion during fermentation, thereby providing a new production method of fixed live yeast and an application device thereof.

Replacement page (Article 26) The task of the invention is achieved as follows:

The present invention provides a method for fixing a living yeast using the chemical principle of the reaction of soluble alginate and soluble calcium salt to produce calcium alginate gel precipitation, so that the soluble calcium salt solution penetrates into the inner cavity of the sponge network, so that the soluble alginate and live yeast The mixed solution forms the backbone network carrier structure of the calcium alginate gel in which the live yeast is embedded in the sponge. The use efficiency and life of the carrier are effectively improved, and a suitable fermentation device is designed for this carrier.

The method for fixing live yeast of the present invention is: putting a live yeast culture solution with a concentration of 0.8 ~ ί / liters into a soluble alginate solution with a concentration of Q.5-5% by weight, and the mixing ratio of the two 5% -40%. After mixing and stirring, put at least one foam sponge into the mixed solution of the above live yeast and soluble alginate solution to make the mixed solution infiltrate; generally immerse 5-30 in the inner cavity space of each cotton. Minutes; then, take out the sponge with the mixed solution and put it into a soluble calcium salt solution with a concentration of 2% to 10% by weight. At this time, a precipitate forms in the internal cavity of the sponge, and the precipitate is seaweed embedded with live yeast. The calcium acid gel (2) is fully gelled after 10 minutes and 20 hours. The cavity inside the sponge is filled with calcium alginate gel embedded with live yeast, and the fixed carrier of the fixed live yeast can be taken out ( Sponge) and ready to use. When the concentration of soluble alginate solution is less than 0.5% by weight, it is difficult to form a gel, and when it is higher than 5% by weight, it is difficult to dissolve. Slow, higher than 10% weight ratio, has toxic effects on live yeast. When the mixed solution of sodium alginate and live yeast penetrates into the inner cavity space of the sponge (1), put the sponge (|) containing the mixed solution into the soluble Among the calcium salt solutions, soluble alginate + soluble calcium salt-calcium alginate gel i + salt is produced.

A preferred solution of the method for fixing live yeast of the present invention is: On the basis of the above, the soluble alginate is sodium alginate, the solution concentration thereof is a weight ratio, the soluble calcium salt is calcium chloride, and the solution concentration thereof is 2% to 10 %weight ratio. For example: The full gelation time of a square sponge with a mixed solution of 8 cm ³ is about 2 hours. Yeast varieties include: Taiwan Yeast No. (F-396), AS. 2. I I89 AS. 2. I 190, Russ ;! (Ra sse II), German No. 2 yeast, Russ (Rass XII, German No. 12 yeast, K-shaped yeast, Nanyang yeast No. 5 (Gate 00), Nanyang mixed yeast, R se se yeast, yeast culture broth The concentration is 100 million pieces / liter.

The immobilized live yeast carrier produced by the method for fixing live yeast provided by the present invention has the following features: the network cavity of the foam sponge body is filled with calcium alginate gel embedded with the live yeast. at this time,

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Replacement page (Article 26) The sexually hardened calcium alginate gel supports the network cavity of the sponge, and in the network space, the calcium alginate gel (2) embedded with live yeast has a network skeleton. The skeleton supports the sponge like reinforced concrete. However, there is a certain gap between the sponge network cavity wall and the gel, and the fermentation broth can freely enter and exit the carrier through the gap, so that the fermentation of live yeast can be fully exerted from the surface to the inside of the carrier. In this way, the sponge hardened because it is supported by the gel network skeleton has strong impact resistance and abrasion resistance, and protects the live yeast from being lost due to impact and friction. For the needs of different fermentation processes, different numbers and different shapes and sizes of carriers can be selected. In addition, the carrier material used in the invention is easy to cut, and the hardness of the gel does not greatly affect the cutting. It is not difficult to cut or cut after making the carrier. It can be easily processed into various shapes according to the fermentation process of various products.

The live yeast immobilization application device of the present invention is: a cage with a mesh, and the prepared immobilized live yeast carrier is loaded into the cage, (the number, shape and size of the carrier (3) are according to the fermentation process Depending on the specific requirements). The cage is fixedly installed in the middle of the inner cavity of the fermentation tank shell. The upper part of the carcass has a feeding opening and a carbon dioxide gas discharge hole, and a lower part has a discharge port. The mesh of the cage (4) is designed to be smaller than the corresponding loading volume, the feeding opening, the discharging opening, the feeding of the fermentation raw materials (such as molasses, etc.) during fermentation, and the output of the finished products (such as alcohol, i, gas) The exhaust hole is used to exhaust the CO ₂ gas generated on the carrier. In addition, for the convenience of maintenance, an upper-access door with a cover can be opened in the upper part of the bath. In addition to the above, the device is used for The fermentation process is exactly the same as the traditional method. For generating excess heat during the fermentation process, you can use the method of spraying cooling water outside the carcass. Other traditional methods can also be used. For example: a spiral cooling pipe in a cage , The carcass is made of a sandwich type, several pipes are connected in parallel on the inner wall of the book.

The advantages of the present invention are-I. Advanced technology. First of all, it has strong resistance to infectious bacteria, which overcomes the shortcomings of frequent fermentation of free fermentation. In the new process, only one batch of carrier strains is used, and one pressing season (4 to 6 months) can be used without changing seeds, thereby reducing the economic loss caused by bacterial conversion. (If the starch alcohol factory uses the carrier to produce, Available for one year).

Using the live yeast fixed carrier of the present invention, the fermentation rate is fast and efficient, and the fermentation cycle only needs 16 to 24 hours (depending on the proportion of the carrier, the amount of the carrier is large, the cycle is short, and the amount of the carrier is small,

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Replacement page (by-law 26 糸) The cycle is slightly longer. The fermentation period of starch alcohol is 24-hours), which is 30% -150% shorter than the traditional free process, and the daily amount of molasses (or starch) is increased by 3Q%-%. The average content of mature crickets is 9.8%-10.7% (ν / ν), which can be up to I |% _ 12% (v / vl; sugar is low, and the amount of molasses alcohol residual sugar is generally between I.2% -1.7 %, Starch alcohol in 0.1% -0.5%.

3. Less investment, easy management, low labor intensity, easy to popularize and apply. The new fermentation tank can save investment by 20% -30%, and the use of the old tank can increase the production capacity by 30% -40%.

4. High economic benefits.

Brief description of the drawings

The present invention is described in detail below with reference to the drawings and embodiments:

0 I is a schematic cross-sectional view of the three fixed-type live yeasts of the present invention, namely A, B, and C.

Figure: It is a schematic longitudinal sectional view of a device using a live yeast fixed load of the present invention.

The illustration is as follows:

1 ― cavernous body 1 ― calcium alginate gel 3 live yeast fixed carrier

4 — cage 5 — fermenter carcass 6 — feed inlet

7 ― Outlet hole 8 ― Outlet port q ― Cooling pipe

The best way to implement the invention

Example 1: (Using sugar cane molasses to make alcohol 1

Production method of live yeast immobilized carrier (3!), Such as ES I—A

Take 20G ml of live yeast (AS 21189 variety) culture solution (the concentration of live yeast culture solution is 1 liter of yeast / liter I; take a piece of artificial foam and cut it into a square square sponge (|) block of ΙΟΟΰ em ³ Several pieces: Add the culture solution to 1QQ ml of sodium alginate solution with a concentration of .5% by weight, stir well, and then immerse the sponge (I) in the mixed solution of sodium alginate and live yeast to make the mixed solution penetrate the sponge Network cavity, soak for 10 minutes and then take out the sponge containing the mixed solution (11 immersed in IG for 5 hours in a calcium chloride solution with a concentration of 5% by weight; work out the sponge that has formed the immobilized live yeast carrier (;) (I), wash with water for later use. At this time, the structure of the carrier U) is: The network cavity in the foam sponge (|) is filled with calcium alginate gel (2) in which live yeast is embedded.

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Change pages carefully (Article 26) Example 2: A fermentation device using the above carrier (]) was prepared, as shown in FIG. 2: A stainless steel cage (4) with a mesh, with a volume of 30M ^{3 was} filled with the above load ^ (3), and The cage (4) is installed in the middle of the fermentation tank housing (5) and fixed with a bracket. The fermentation tank carcass (5) is a cylindrical body with a volume. There is a feed inlet W and a C0 ₂ gas exhaust hole (7) on the upper part, and a covered door for people to enter and repair. The lower part has a discharge port: a stainless steel cooling pipe (9) has two ends passing through A sleeve hole on the wall of the fermenter carcass (5), which is welded and sealed to the pipe (9). The pipe (9) penetrates and is placed in a cage (4) in a spiral shape. Its role is to remove excess heat generated during fermentation on the load and take it away through the cooling water flowing in the pipe (9). The fermentation temperature is generally 10'C-40'C.

Example (production of glycerin)

Except for using a special technology for producing glycerol by fermentation of starch hydrolyzed sugar, the rest are the same as in Example 1.

Example 4: Preparation of a fixed live yeast carrier (3), take Taiwan Yeast No. 96 (F-396) 40ml (concentration of 〖100 million yeasts / ml), add the live yeast culture solution to a concentration of 0. S 20% ml of sodium alginate solution by weight, stir and mix well, soak at room temperature for 10 minutes, then take out the sponge (I) that is saturated with the mixture and immerse it in a calcium chloride solution with a concentration of 10% by weight Five hours, a gel calcium alginate was embedded in the sponge cavity to embed the live yeast carrier.

Glycerol was produced by fermentation of starch-hydrolyzed sugar using the system shown in Figure 2.

Example 5: The rest of the process is the same as in Example I, and R is a live yeast (Ras s e H i German No. 2 yeast).

Example 6: (Alcohol fermentation by starch hydrolysis with sugar)

The live yeast used was Nanyang No. 5 (Gate i or Nanyang mixed yeast (Gate; its yeast culture solution concentration was 100 million per liter and sodium alginate solution (concentration 5% 1, and its mixing ratio was | 5%). The concentration of the calcium chloride solution used is 5%, and the soaking time is Π minutes. The prepared live yeast carrier is used to ferment alcohol by using starch hydrolysis sugar.

Example 7: The same process as in Example I, except that beer yeast was used.

After the fixed live yeast is prepared, the beer is prepared by the conventional beer fermentation process.

EXAMPLES The method for fixing yeast was the same as in Example 1, except that AS 2 1190 yeast was used.

Claims

Claim

1. A method for fixing live yeast, characterized in that: a concentration of live yeast of 80-200 million per liter of live yeast is added to a concentration of G. 5% -5% by weight soluble alginate solution The ratio of the live yeast culture solution to the soluble alginate solution is from 5% to 40%. Stir and mix well, and put at least one clean foam sponge (I) into the mixture of the live yeast and the soluble alginate solution. , Soak for 5-30 minutes to allow the mixed solution to penetrate into the inner cavity space of the sponge U), and then, take out the sponge (I) with the mixed solution and place it at a concentration of 2 ° /. -Soluble calcium salt solution in% by weight, gelation at l (rc-4 () ° C, reaction time is 1 minute-10 hours, wait for live yeast to be embedded in sponge (li After the calcium alginate has been coagulated, remove the sponge that has become a live yeast fixation carrier (3).

2. The method for fixing live yeast according to claim 1, wherein the soluble alginate is sodium alginate and potassium alginate.

3. The method for fixing live yeast according to claim 1 or 2, characterized in that: the immobilized live yeast carrier is filled with embedding in the foam network cavity in the foam sponge (I) Alginate gel from live yeast (2) ο

4. The application device of the method for fixing live yeast according to claim 1 or claim 5, characterized in that: the immobilized live yeast carrier (3) is packed into a cage (4i) with a mesh, and the cage (4) is fixedly installed in the fermentation The middle part of the inner cavity of the tank shell (5), the upper part of the shell (5) has a feed opening (0), and a carbon dioxide gas discharge hole, and a lower part has a discharge port).

5. The application device of the method for fixing live yeast according to Forklift claim 4, characterized in that: it comprises installing a cooling pipe (?> In the fermenter, and both ends of the cooling pipe w pass through the wall of the fermenter shell respectively The sleeve hole is tightly connected with the pipe (9).

6. The application device of the method for fixing live yeast according to claim 4, characterized in that the cooling pipe (9) comprises a spiral tube shape, a carcass body (5), and a plurality of pipes are sandwiched into a sandwich type. Parallel to the inner wall of the casing (5).

7. The method for fixing live yeast according to claim 1, wherein the soluble calcium salt is calcium chloride.

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Replacement page (Article 26)