CN108315230B - Reactor and method for preparing yeast culture by using reactor - Google Patents
Reactor and method for preparing yeast culture by using reactor Download PDFInfo
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- CN108315230B CN108315230B CN201810199308.1A CN201810199308A CN108315230B CN 108315230 B CN108315230 B CN 108315230B CN 201810199308 A CN201810199308 A CN 201810199308A CN 108315230 B CN108315230 B CN 108315230B
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 33
- 238000000855 fermentation Methods 0.000 claims abstract description 23
- 230000004151 fermentation Effects 0.000 claims abstract description 23
- 238000012258 culturing Methods 0.000 claims abstract description 20
- 238000009423 ventilation Methods 0.000 claims abstract description 6
- 239000001963 growth medium Substances 0.000 claims description 29
- 239000002518 antifoaming agent Substances 0.000 claims description 28
- 238000003756 stirring Methods 0.000 claims description 28
- 238000010298 pulverizing process Methods 0.000 claims description 16
- 238000004519 manufacturing process Methods 0.000 claims description 9
- 239000002609 medium Substances 0.000 claims description 9
- 230000003698 anagen phase Effects 0.000 claims description 5
- 239000013530 defoamer Substances 0.000 claims description 5
- 241000894007 species Species 0.000 claims description 4
- 239000007789 gas Substances 0.000 description 37
- 230000008569 process Effects 0.000 description 17
- 239000007788 liquid Substances 0.000 description 15
- 238000009630 liquid culture Methods 0.000 description 15
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 9
- 239000001301 oxygen Substances 0.000 description 9
- 229910052760 oxygen Inorganic materials 0.000 description 9
- 230000001954 sterilising effect Effects 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 235000015067 sauces Nutrition 0.000 description 5
- 210000005253 yeast cell Anatomy 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 239000006260 foam Substances 0.000 description 4
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 4
- 230000000630 rising effect Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
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- 239000008103 glucose Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 230000002035 prolonged effect Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
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- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 235000019764 Soybean Meal Nutrition 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 239000012267 brine Substances 0.000 description 2
- 239000000498 cooling water Substances 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- 239000004455 soybean meal Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
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- 239000010779 crude oil Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
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- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/02—Means for regulation, monitoring, measurement or control, e.g. flow regulation of foam
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Organic Chemistry (AREA)
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- Zoology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
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Abstract
The invention provides a reactor and a method for preparing yeast culture by using the reactor. Wherein the reactor comprises: a tank body; the gas distributor is arranged in the tank body and can be communicated with an external gas source, and a plurality of holes are distributed on the gas distributor; and a bubble crushing tray disposed above the gas distributor. The method for preparing the yeast culture by using the reactor comprises the following steps: performing expansion culture on the strain; inoculating the strain subjected to the expanded culture into the reactor for culture, wherein the gas distributor is used for uniformly distributing gas while ventilation is carried out in the reactor, and the bubble crushing disc is used for bubble crushing; and inoculating the strain cultured in the reactor into a fermentation tank for culturing to obtain a yeast culture.
Description
Technical Field
The invention relates to the fields of bioengineering, chemical engineering and environmental engineering, in particular to a reactor and a method for preparing a yeast culture by using the reactor.
Background
Yeast cultures are micro-ecological products formed by fermenting yeast sufficiently on a specially made medium under the control of specific process conditions. The general production process of the yeast culture mainly comprises a strain expansion culture stage and a fermentation tank fermentation stage, wherein the conventional strain expansion culture stage mainly comprises the stages of slant culture, triangular flask culture, karsch tank culture, seed tank culture and the like of original yeast strains. The seed tank is a device for culturing seeds by utilizing microorganisms, is suitable for fermentation, brewing, seasoning, pharmacy, chemical industry and fungus cultivation of yoghurt and lactobacillus beverage mother liquor, and has the functions of heating, heat preservation and cooling, and raw materials in the tank are mixed through a stirring system.
When the seed tank is used for culturing the strain of the yeast culture, the strain is usually required to be subjected to oxygen-introducing stirring culture, and a single air inlet pipe which is directly introduced into the bottom of the seed tank body is arranged in the seed tank body in the prior art, and compressed air realizes the oxygen-introducing operation on the strain cultured in the seed tank body through the air inlet pipe. In the usual case, without major concentration-enhancing changes in the ingredients (culture substrates) in the seed tankThe seed tank needs 3-4 days of production period, and the quality index of mature yeast seed liquid is that the number of yeast cells is 2.5-4×10 8 In order to improve the requirements of mature seed liquid indexes, the mature cells (colony monomers) also need longer culture time or change the nutrient components of the culture medium, so that the culture time is prolonged, more compressed gas needs to be introduced to cause low production efficiency and low equipment utilization rate, and meanwhile, the production cost is obviously improved by increasing the content of the nutrient components.
The matters in the background section are only those known to the inventors and do not, of course, represent prior art in the field.
Disclosure of Invention
In one aspect of the present invention, there is provided a reactor comprising:
a tank body;
the gas distributor is arranged in the tank body and can be communicated with an external gas source, and a plurality of holes are distributed on the gas distributor; and
and the bubble crushing disc is arranged above the gas distributor.
According to one aspect of the invention, wherein the gas distributor is an annular duct.
According to one aspect of the invention, the opening direction of the hole is 45 degrees relative to the plane of the pipeline.
According to one aspect of the invention, wherein the diameter of the hole is 4-5mm.
According to one aspect of the invention, wherein the bubble shredder plate is rotatable.
According to one aspect of the invention, wherein the bubble smashing disc is a disc, and the diameter of the disc is 1/2-2/3 of the diameter of the reactor tank body.
According to one aspect of the invention, a plurality of air holes are distributed on the bubble smashing disc, and the diameter of each air hole is 10-15mm.
According to one aspect of the invention, the bubble breaking disc is further provided with a plurality of bubble breaking teeth.
According to one aspect of the invention, the number of the bubble crushing teeth is 6, the included angle between every two adjacent bubble crushing teeth is 60 degrees, and meanwhile, the included angle between the crushing teeth and the plane of the disc is 60 degrees.
According to one aspect of the invention, the reactor further comprises a motor, a stirring shaft and a stirring paddle, wherein the stirring paddle is positioned in the tank body, fixedly installed on the stirring shaft and can rotate along with the stirring shaft, and the motor can drive the stirring shaft to rotate.
According to one aspect of the invention, wherein the bubble breaker plate is fixedly mounted on the agitator shaft.
In another aspect of the invention, the invention provides the use of the reactor described above as a seed tank.
In another aspect of the present invention, there is provided a method for preparing a yeast culture using the above-described reactor, comprising:
performing expansion culture on the strain;
inoculating the strain subjected to the expanded culture into the reactor for culture, wherein the gas distributor is used for uniformly distributing gas while ventilation is carried out in the reactor, and the bubble crushing disc is used for bubble crushing; and
and inoculating the strain cultured in the reactor into a fermentation tank for culturing to obtain a yeast culture.
According to one aspect of the invention, the step of performing the expansion culture on the strain comprises one or more of the following:
inoculating strain into the slant/plate culture medium to obtain slant/plate culture strain;
inoculating the slant/flat plate culture strain into a culture medium in a triangular flask for culturing to obtain a triangular flask culture strain; and
and inoculating the triangular flask culture strain into a culture medium in a Karsch tank for culturing to obtain the Karsch tank culture strain.
According to one aspect of the present invention, the method for producing a yeast culture further comprises adding an antifoaming agent to a culture medium before sterilization in the reactor before the culturing of the strain after the expansion culture in the reactor; and/or adding an antifoaming agent to the reactor during the culturing in the reactor.
According to one aspect of the invention, wherein the amount of antifoaming agent added to the medium before sterilization in the reactor is 20-30% of the total amount of antifoaming agent, wherein the total amount of antifoaming agent is 0.4-1.0% of the total amount of yeast culture. Preferably, the antifoaming agent is added to the reactor during the cultivation in the reactor in an amount of 70-80% of the total amount of the antifoaming agent.
According to one aspect of the invention, wherein the adding of the antifoaming agent to the reactor during the culturing in the reactor is performed in multiple times before the logarithmic growth phase of the strain.
According to one aspect of the invention, wherein said adding said antifoaming agent in multiple portions prior to the logarithmic growth phase of said bacterial species is performed as follows: 1 ratio the remaining 70-80% of defoamer was added in two portions.
In another aspect of the invention, a yeast culture is provided, prepared according to the method of preparing a yeast culture described above.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
FIG. 1 is a schematic illustration of a reactor according to one embodiment of the invention;
FIG. 2 is a schematic view of a gas distributor according to one embodiment of the present invention;
FIG. 3 is a schematic view of a gas distributor according to one embodiment of the present invention;
fig. 4 is a schematic bottom view of a bubble reducing tray provided with bubble reducing teeth in accordance with one embodiment of the present invention.
Detailed Description
Hereinafter, only certain exemplary embodiments are briefly described. As will be recognized by those of skill in the pertinent art, the described embodiments may be modified in various different ways without departing from the spirit or scope of the present invention. Accordingly, the drawings and description are to be regarded as illustrative in nature and not as restrictive.
In the description of the present invention, it should be understood that the directions or positional relationships indicated by the terms "upper", "lower", "bottom", "inner", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of description and simplification of the description, and do not indicate or imply that the apparatus or element in question must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, in the description of the present invention, the meaning of "a plurality" means one, two or more than two, unless explicitly defined otherwise.
In the description of the present invention, it should be noted that the term "mounted" should be interpreted broadly, unless explicitly stated and defined otherwise, such as for example, either fixedly mounted, detachably mounted, or integrally mounted. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.
The following disclosure provides many different embodiments, or examples, for implementing different features of the invention. In order to simplify the present disclosure, components and arrangements of specific examples are described below. They are, of course, merely examples and are not intended to limit the invention. Furthermore, the present invention may repeat reference numerals and/or letters in the various examples, which are for the purpose of brevity and clarity, and which do not themselves indicate the relationship between the various embodiments and/or arrangements discussed. In addition, the present invention provides examples of various specific processes and materials, but one of ordinary skill in the art will recognize the application of other processes and/or the use of other materials.
The preferred embodiments of the present invention will be described below with reference to the accompanying drawings, it being understood that the preferred embodiments described herein are for illustration and explanation of the present invention only, and are not intended to limit the present invention.
As shown in fig. 1, in a first embodiment of the present invention, a reactor 100 is provided. The reactor 100 includes: a tank 1, a gas distributor 2 and a bubble-pulverizing tray 3. The gas distributor 2 (the detailed structure will be described below) is disposed in the tank 1, and in the preferred embodiment of the present invention, the gas distributor 2 is disposed at the bottom of the tank 1, and those skilled in the art can also contemplate that the gas distributor 2 is disposed at other positions in the tank 1, which are all within the scope of the present invention. The gas distributor 2 may be in communication with an external gas source, for example an external air source (or a compressed air source) to effect ventilation of the interior of the tank 1. The gas distributor 2 may be a pipe and may be provided with a plurality of holes, for example 1, 2, 3 or more than 3 holes. The bubble crushing tray 3 (detailed structure will be described later) is provided above the gas distributor 2.
As shown in fig. 2, a schematic bottom view of the gas distributor 2 of the present invention is shown, according to a preferred embodiment of the present invention. The gas distributor 2 may be an annular pipe, such as an annular circular pipe or an annular square pipe, and in this embodiment, is shown as an integrally connected pipe, and it should be understood by those skilled in the art that the pipe may also be an open-ended pipe and a closed-ended pipe, as shown in fig. 3, and the closed position may also be disposed at any position of the pipe, which is within the scope of the present invention. A plurality of holes 21 may be uniformly distributed on the gas distributor 2, and the diameter of the holes 21 may be 4-5mm. According to a preferred embodiment of the present invention, the opening direction of the hole 21 may be that the plane of the pipe is 45 degrees downward, which is more beneficial to prolonging the residence time of the gas in the tank 1 after the gas introduced from the gas inlet pipe 6 is sprayed out through the gas distributor 2, so that the oxygen is better absorbed.
As shown in fig. 4, a schematic bottom view of the bubble breaker tray 3 of the present invention is shown, according to a preferred embodiment of the present invention. The bubble-pulverizing disc 3 may be a disc having a diameter of 1/2-2/3 of the diameter of the reactor tank 1, and it should be understood by those skilled in the art that the bubble-pulverizing disc 3 may be a square disc, an oval disc, etc., which are within the scope of the present invention. A plurality of air holes, for example, 1, 2, 3 or more than 3 air holes, can be uniformly arranged on the bubble crushing disc, and the diameter of the air holes can be 10-15mm. According to a preferred embodiment of the present invention, as shown in fig. 4, the bubble-pulverizing disc 3 may further be provided with a plurality of bubble-pulverizing teeth 4, for example, 2, 3, 4 or more than 4, in this embodiment, 6 bubble-pulverizing teeth 4 are shown, and an included angle between two adjacent bubble-pulverizing teeth may be 60 degrees, which is understood by those skilled in the art that an included angle between two adjacent bubble-pulverizing teeth may be any angle, and these are all within the scope of the present invention. According to a preferred embodiment of the invention, the bubble breaking teeth 4 are at an angle of 60 degrees to the bubble breaking disc 3. The bubble breaking teeth 4 may for example be narrow strips on which teeth are distributed. The bubble breaking teeth 4 may be provided on the bubble breaking disc 3 by means of welding, and other connection means, such as screw connections, are conceivable by a person skilled in the art, which are within the scope of the invention. According to a preferred embodiment of the invention, the bubble-pulverizing disc 3 or the combination of the bubble-pulverizing disc 3 and the bubble-pulverizing teeth 4 may be rotatable, e.g. by arranging the bubble-pulverizing disc on a spindle in the tank 1 to effect rotation thereof. Compressed gas sprayed out from the gas distributor 2 at the bottom of the tank body 1 can generate relatively violent stirring in the upward turning process, and the generated large bubbles can be crushed, stirred and sealed and mixed through the redistribution and buffering of the bubble crushing disc 3 and the bubble crushing teeth 4, so that the large bubbles are changed into smaller bubbles, the rising process is more uniformly distributed in the tank body 1, the rising time of the compressed gas is greatly prolonged, and the possibility that oxygen in the compressed gas is absorbed in the culture process is higher. Thus, the same oxygen supply amount can increase the amount of oxygen dissolved in the culture solution in the tank body 1 during the culture process, thereby being more beneficial to the culture, reducing the supply amount of compressed gas and saving the supply of the compressed gas.
According to a preferred embodiment of the present invention, as shown in fig. 1, the reactor 100 provided by the present invention further comprises a motor 7, a stirring shaft 9 and a stirring paddle 10. Wherein the stirring paddle 10 is positioned in the tank body 1, is fixedly arranged on the stirring shaft 9 and can rotate along with the stirring shaft 9, and the motor 7 can drive the stirring shaft 9 to rotate. Those skilled in the art will appreciate in view of this disclosure that multiple layers of paddles 10 may be alternatively employed, such as two layers, three layers, etc., and that paddles 10 may alternatively be employed in conventional anchor, paddle, turbine, push or frame configurations, etc. According to a preferred embodiment of the present invention, the bubble-pulverizing disc 3 or the combination structure of the bubble-pulverizing disc 3 and the bubble-pulverizing teeth 4 may be mounted on the stirring shaft 9, for example, on the lower end of the stirring shaft 9, for example, by means of a screw bolt, so as to be mounted on the lower end of the stirring shaft 9, and thus may rotate together with the stirring paddle 10 along with the rotation of the stirring shaft 9, thereby achieving better stirring of the gas in the culture solution in the tank 1. As shown in fig. 1, the reactor 100 provided by the invention further comprises an air inlet 5 and an air inlet pipe 6, wherein the air inlet 5 is used for communicating with an external air source, and the external air source is conveyed into the gas distributor 2 through the air inlet pipe 6 and used for ventilating the culture solution in the tank body 1. As shown in fig. 1, the reactor 100 provided by the present invention may further include a speed reducer 8, a cooling water inlet 11, a cooling water outlet 12, a baffle 13, a coil 14, a heat insulation layer 15, a tank inlet 16, a feed inlet 17, a discharge outlet 18 and a support leg 19, and those skilled in the art can reasonably modify these components according to the disclosure and the prior art, which are all within the scope of the present invention.
In a second embodiment of the present invention, the present invention provides a method for preparing a yeast culture using the reactor 100 disclosed herein. The method comprises the following steps:
performing expansion culture on the strain;
inoculating the strain subjected to the expanded culture into the reactor 100 provided by the invention for culture, wherein the gas distributor 2 is utilized to uniformly distribute gas while ventilation is carried out in the reactor 100, and the bubble crushing and capping are carried out by utilizing the bubble crushing disc 3 or the combined structure of the bubble crushing disc 3 and the bubble crushing teeth 4; and
the strain cultured in the reactor 100 is inoculated into a fermenter to be cultured, thereby obtaining a yeast culture.
According to a preferred embodiment of the invention, the step of performing an expanded culture of the species comprises one or more of the following:
inoculating strain into the slant/plate culture medium to obtain slant/plate culture strain;
inoculating the slant/flat plate culture strain into a culture medium in a triangular flask for culturing to obtain a triangular flask culture strain; and
and inoculating the triangular flask culture strain into a culture medium in a Karsch tank for culturing to obtain the Karsch tank culture strain.
According to a preferred embodiment of the present invention, the above slant/plate medium comprises the following components in parts by weight: malt extract powder 30.0g, soybean peptone 3.0g, agar 15.0g, water to 1000mL, pH 5.6+ -0.2 (the weight of the above components only represents the weight ratio among the components, not the actual mass). Heating the slant/plate culture medium to dissolve each component, packaging into several test tubes or plates, sealing test tubes or plates, sterilizing at 121deg.C for 30min, taking out test tubes, placing on slant or plates, and cooling. Placing the slant/plate culture medium in a drying oven or an incubator for 48 hours for aseptic test, inoculating yeast strains prepared in advance, inoculating 2-3 platinum fungus into each slant test tube or plate, and culturing at 30+ -1deg.C for 96 hours to obtain slant/plate culture strain.
According to a preferred embodiment of the invention, a triangular flask liquid culture medium is prepared according to the formula requirement, and the liquid culture medium comprises the following components in parts by weight: glucose 35-50g; sauce: 150-200ml (wherein, the salt content is 14-18g/100ml, and the total nitrogen is 1.5-1.7g/100 ml); brine: 250-300ml (salt content 22-23g/100 ml); adding water to a volume of 1000mL (ensuring that the nitrogen content is less than or equal to 0.25g/100mL, the sodium chloride is less than or equal to 10g/100mL, and the glucose is less than or equal to 5%). The sauce is prepared from defatted soybean meal and wheat flour in a ratio of 1:0.8 by a starter propagation process, and is prepared from the following raw materials: salt water is added at a ratio of about 1:2.2.+ -. 0.1 (w/w) to the concentrationBrine with the concentration of 22+/-0.1 g/100ml is mixed into a fermentation cabin, wherein yeast liquid (which can be, for example, previous fermented mash, is squeezed to obtain juice after fermentation for 6 months, is recycled, and is used as sauce after fermentation, flavoring yeast liquid is added according to the process requirement in the fermentation process of the fermented mash for flavoring) with the proportion of 1% is added, and after fermentation for 6 months, sauce mash is formed, and the sauce is obtained after squeezing. The prepared triangular flask liquid culture medium is divided into a plurality of 500mL triangular flasks, for example, 200-300mL liquid culture medium can be added into each triangular flask, and the pH value is regulated to 4.5-5.1; sealing, sterilizing at 121deg.C for 30min, and cooling. Under the aseptic condition, the liquid culture medium in each triangular flask is inoculated with the prepared slant/plate culture strain, for example, platinum ears or an inoculating loop can be used for transplanting the slant/plate culture strain to the liquid culture medium in the triangular flask for 2-3 times, so that the number of yeast strains in the liquid culture medium in the triangular flask reaches 5-8 x 10 4 About one/ml. Shake cultivation can be carried out on the triangular flask, the temperature is controlled to be 30+/-1 ℃, and the rotating speed can be 200-300r/min; culturing for 72-96h by shaking to obtain the triangular flask culture strain.
According to a preferred embodiment of the present invention, after the preparation of the triangular flask culture strain, the triangular flask culture strain may be completely inoculated into a casserole for culturing, so that the number of yeast cells in the liquid medium in the casserole reaches 2-5×10 6 About one volume/ml, the liquid culture medium in the Karsch tank can be the same as that in a triangular flask, for example, 20-30L of the liquid culture medium can be added into the Karsch tank, the pH value is regulated to 4.5-5.1, the liquid culture medium is aerated and cultured for 48-72 hours under the aseptic culture condition, the temperature is controlled to 30+/-1 ℃, and the aeration speed is controlled to be 0.1-0.3m 3 And (3) preparing a Karsch tank culture strain, wherein mature liquid indexes are as follows: the cell number is more than or equal to 2 multiplied by 10 8 The method comprises the steps of carrying out a first treatment on the surface of the Germination rate is 20-30%; the miscellaneous bacteria is less than or equal to 100cfu/ml; the pH is 4.5-5.5.
According to a preferred embodiment of the present invention, after the yeast strains are subjected to the expansion culture, the prepared yeast strains subjected to the expansion culture are transplanted into the reactor 100 provided by the present invention for continuous culture.A reactor liquid culture medium, for example, 2000-3000L of reactor liquid culture medium, is added in advance in the reactor 100, and the reactor liquid culture medium is prepared according to a formula, wherein the reactor liquid culture medium comprises the following components in parts by weight: glucose 75Kg, crude oil (total nitrogen: 1.5g/100ml, sodium chloride: 16.5g/100 ml): 330L, prepared saline (concentration: 22.0g/100 ml), 660L, constant volume to 2000L, and pH adjusted to 4.5-5.1. Sealing, sterilizing at 121deg.C for 30min, or indirectly steam sterilizing at 100-105deg.C for 25-40min, and maintaining the temperature for 20-30min. Cooling to 30+ -1deg.C, inoculating the strain after the expansion culture to make the yeast inoculation amount about 1% of the total weight of the mixture in the reactor 100, and cell concentration reaching: 1-3*10 6 And each ml. According to a preferred embodiment of the present invention, an antifoaming agent may be added to the liquid medium in the reactor 100 before the liquid medium in the reactor 100 is sterilized such that the weight content of the antifoaming agent is 0.4 to 1.0%. The invention preferably adopts 20-30% of the total amount of the defoamer before sterilization, can ensure that the defoamer is fully and uniformly mixed after being heated to form fine liquid drops, so as to ensure that the foam overflow phenomenon can not occur for a long time after the strain after the expansion culture is inoculated into the reactor. After the strain subjected to the expanded culture is connected into the reactor 100 according to the proportion of 1 (75-100), as shown in fig. 1, a tank body inlet hole 16 and a feed inlet 17 of the reactor 100 are closed, a motor 7 is started to stir, meanwhile, a valve of an air inlet 5 and a valve of a filter (not shown in fig. 1) are opened to enable an external air source to be sprayed into the reactor 100 through the filter, an air inlet pipe 6 and an air distributor 2 after purification treatment, relatively violent stirring can be generated in the air upturning process, the generated large bubbles can be crushed, stirred and sealed and mixed through a bubble crushing disc 3 and a bubble crushing tooth 4, so that the large bubbles are changed into smaller bubbles, the rising process is more evenly distributed in the tank body 1, the rising time of the large bubbles is greatly prolonged, and the possibility that oxygen in the compressed air is absorbed in the culture process is higher. Thus, the same oxygen supply amount can increase the amount of oxygen dissolved in the culture solution in the tank 1 during the culture process, thereby being more beneficial to the culture and simultaneouslySo as to reduce the supply amount of the compressed gas and save the supply of the compressed gas. Wherein the ventilation is set to 10-20m 3 And/h, performing automatic heat-preserving culture at 30+/-1 ℃ for 40-48h to obtain a reactor culture strain, wherein mature liquid indexes are as follows: the cell number is more than or equal to 5 multiplied by 10 8 The method comprises the steps of carrying out a first treatment on the surface of the Germination rate is 20-30%; the miscellaneous bacteria is less than or equal to 100cfu/ml; the pH is 4.5-5.5.
According to a preferred embodiment of the invention, an antifoaming agent may be added to the mixed solution of the reactor during the cultivation in the reactor, for example, the antifoaming agent may be added in a plurality of times before the logarithmic growth phase of the strain in the reactor, for example, the antifoaming agent may be added in two times in a ratio of 1:1, and the amount of the antifoaming agent added in the process may be 70-80% of the total amount of the antifoaming agent added to the reactor, so as to ensure that the strain does not overflow foam during the cultivation in the reactor 100. The invention adopts the liquid culture medium in the reactor 100 to be added with the defoamer for times before and after sterilization, which can fully ensure that the yeast strain is always covered by foam in the early culture process, but no foam overflows, thereby increasing more dissolved oxygen opportunities and reducing the introduced oxygen from rapidly falling out of the liquid surface to be discharged. Note that the present invention is not limited to the necessity of adding the antifoaming agent twice, and for example, the antifoaming agent may be added to the liquid medium in the reactor 100 just before the liquid medium in the reactor 100 is sterilized so that the weight content of the antifoaming agent is 0.4 to 1.0%.
According to a preferred embodiment of the present invention, after the reactor culture strain cultured in the reactor 100 is prepared, it is put into a fermenter to ferment. After fermenting the fermented mash in the fermentation tank for 30-40d in advance, heating the fermented mash, wherein the heating range is ensured to be about 1 ℃/day, and finally, slowly heating the fermentation tank to 22-30 ℃ and controlling the pH value to 5.0-5.5, and then, putting the yeast strain cultivated by the reactor into the fermentation tank for fermentation. Wherein the fermented mash is prepared from defatted soybean meal and wheat flour according to a ratio of about 1:0.8 by a starter propagation process, and the following raw materials: brine with the concentration of 22+/-0.1 g/100ml is added in the proportion of about 1:2.2+/-0.1 (w/w) and mixed into a fermentation tank for fermentation. Reactor cultures prepared by using the reactor disclosed by the inventionThe yeast cell number in the fermented mash can reach 5×10 by adding the strain into the fermentation tank according to the addition amount of about 1% 6 The addition amount of the yeast liquid is 3-5% of the amount of the fermented mash, and the number of yeast cells in the fermented mash reaches 10 5 --10 6 General technological requirements of about one per ml. The large number of yeast cells can be propagated more quickly at the beginning of fermentation, dominant thalli can be formed quickly in the fermentation process, the growth, propagation and metabolism of other miscellaneous bacteria are inhibited, other miscellaneous substances generated by other miscellaneous bacteria are inhibited, and the flavor substances generated by fermentation are ensured to be carried out towards the direction required by people.
In the process of preparing yeast culture by using the reactor provided by the invention, under the condition that ingredients in the reactor are unchanged, the index of the cell number of mature yeast liquid is improved by 40-70% and is increased to 5.5-6 multiplied by 10 after culturing for 40-48 hours 8 The culture time is shortened to 40-48h compared with the original 72-96h, and the equipment utilization rate is improved by more than 80%. Meanwhile, a small amount of reactors are added into the fermentation tank to cultivate strains, so that the fermentation effect can be achieved, and the production cost is low.
The invention also provides a yeast culture prepared according to the method for preparing the yeast culture.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (15)
1. A reactor, comprising:
a tank body;
the gas distributor is arranged in the tank body and can be communicated with an external gas source, and a plurality of holes are distributed on the gas distributor; and
the bubble crushing dish, bubble crushing dish sets up the top of gas distributor, bubble crushing dish is rotatable, it has a plurality of gas pockets to distribute on the bubble crushing dish, the downside of bubble crushing dish still is provided with a plurality of bubble crushing teeth, bubble crushing teeth follow the radial setting of bubble crushing dish, it has the tooth to distribute on the bubble crushing teeth.
2. The reactor of claim 1, wherein the gas distributor is an annular conduit; and/or the opening direction of the hole is the plane where the pipeline is positioned and is at an angle of 45 degrees; and/or the diameter of the hole is 4-5mm.
3. The reactor of claim 1 or 2, wherein the bubble-pulverizing disc is a disc having a diameter of 1/2-2/3 of the diameter of the reactor tank; and/or the diameter of the air hole is 10-15mm.
4. The reactor of any one of claims 1-2, wherein the bubble crushing teeth are at an angle of 60 degrees to the bubble crushing disc.
5. The reactor of claim 4, wherein the number of bubble crushing teeth is 6, and the included angle between two adjacent bubble crushing teeth is 60 degrees.
6. The reactor of any one of claims 1-2, wherein the reactor further comprises a motor, a stirring shaft, and a stirring paddle, the stirring paddle being located within the tank and fixedly mounted on and rotatable with the stirring shaft, the motor being operable to drive the stirring shaft to rotate.
7. The reactor of claim 6, wherein the bubble disk is fixedly mounted on the agitator shaft.
8. Use of the reactor of any one of claims 1-7 as a seed tank.
9. A method of preparing a yeast culture using the reactor of any one of claims 1-7, comprising:
performing expansion culture on the strain;
inoculating the strain subjected to the expansion culture into the reactor according to any one of claims 1 to 7 for culture, wherein the gas distributor is used for uniformly distributing gas while ventilation is carried out in the reactor, and the bubble crushing disc is used for bubble crushing; and
and inoculating the strain cultured in the reactor into a fermentation tank for culturing to obtain a yeast culture.
10. A method of preparing a yeast culture according to claim 9, the step of performing an expanded culture of the species comprising one or more of:
inoculating strain into the slant/plate culture medium to obtain slant/plate culture strain;
inoculating the slant/flat plate culture strain into a culture medium in a triangular flask for culturing to obtain a triangular flask culture strain; and
and inoculating the triangular flask culture strain into a culture medium in a Karsch tank for culturing to obtain the Karsch tank culture strain.
11. The method for producing a yeast culture according to claim 9, further comprising adding an antifoaming agent to the culture medium in the reactor before inoculating the strain after the expansion culture into the reactor according to any one of claims 1 to 7 for culturing; and/or adding an antifoaming agent to the reactor during the culturing in the reactor of any of claims 1-7.
12. The method for producing a yeast culture according to claim 11, wherein the amount of the antifoaming agent added to the medium in the reactor is such that the weight content of the antifoaming agent is 0.4 to 1.0%o and/or the antifoaming agent is added to the reactor during the cultivation in the reactor in an amount of 70 to 80% of the total amount of the antifoaming agent.
13. The method for producing a yeast culture according to claim 11, wherein the adding of an antifoaming agent to the reactor during the cultivation in the reactor according to any one of claims 1 to 7 is adding the antifoaming agent in a plurality of times before the logarithmic growth phase of the strain.
14. The method of preparing a yeast culture according to claim 13, wherein the adding the antifoaming agent in multiple portions prior to the logarithmic growth phase of the species is performed at a rate of 1: the defoamer was added in two portions at 1 ratio.
15. A yeast culture prepared according to the method of preparing a yeast culture of any one of claims 9-14.
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