CN110684680A - Preparation method of high-density yeast fermentation liquor - Google Patents
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Abstract
The invention relates to a preparation method of high-density yeast fermentation liquor, which comprises the steps of mixing corn bean pulp with water, adding high-temperature amylase, saccharifying enzyme and acidic protein respectively, carrying out thorough enzymolysis, and carrying out solid-liquid separation to obtain solid wet residue and dilute composite nutrient solution. Diluting a part of the dilute composite nutrient solution, inoculating the diluted composite nutrient solution into a yeast seed solution for culturing, and evaporating the rest of the dilute composite nutrient solution under reduced pressure to obtain a concentrated solution. And in the culture process, sufficient dissolved oxygen is controlled by feeding the concentrated composite nutrient solution and the nutrient salt, when the density of the yeast is maximum, feeding is stopped, the yeast fermentation liquid is obtained after the consumption of the fermentable sugar is completely consumed, and the high-density yeast liquid is obtained by standing. The invention is characterized in that different enzyme preparations are enzymolyzed to obtain sugar and amino acid required by yeast growth, and high-density yeast liquid is obtained in a batch discharging mode of continuous fed-batch fermentation, so that the yeast content is improved, the consumption of fermentation sterilization steam is low, the production water is low, the discharge amount of waste water from tank washing of a fermentation tank is reduced, and the effects of energy conservation, consumption reduction and clean production are achieved.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a preparation method of high-density yeast fermentation liquor.
Background
The saccharomyces cerevisiae has high nutritive value of mycoprotein, the protein content in yeast dry matter can reach 50 percent, the content of essential amino acid required by animals is high, the content is close to animal protein, and the utilization rate of net protein is far higher than that of plant protein. In addition, the saccharomyces cerevisiae also contains abundant nucleotide, B vitamins, minerals and other physiological active substances. With the development of microbial engineering and fermentation engineering, saccharomyces cerevisiae is more and more widely used for producing high-quality protein feed.
Meanwhile, with the wide application of yeast in feed, the realization of high-density fermentation of yeast in a fermentation tank space with limited volume is a key for maintaining the production scale of yeast products and controlling the cost, so that research on various characteristics of yeast, particularly fermentation characteristics, is necessary.
According to the traditional yeast value-added fermentation method, high-density fermentation of saccharomyces cerevisiae is realized by optimizing culture conditions, and a set of complete fermentation process is formed by integrating and optimizing a plurality of factors and mutual influences thereof, and the process is a work with complex procedure and high comprehensiveness and is not beneficial to the requirement of industrial development. In addition, the raw materials such as glucose and yeast extract are mostly used as reducing sugar and nitrogen source for yeast culture, and the production cost is high. Therefore, according to the utilization mode of the saccharomyces cerevisiae on reducing sugar and nitrogen source, the yeast high-density culture production process which is simple and convenient to operate, suitable for large-scale industrial production, energy-saving and environment-friendly by adopting raw materials with wide sources and low price is needed to be developed.
Disclosure of Invention
The invention aims to utilize cheap raw materials, carry out enzymolysis on the raw materials into sugar and an organic nitrogen source required by yeast growth through different enzyme preparations, and obtain high-density yeast liquid in a continuous fed-batch fermentation batch discharging mode, so that the yeast content is improved, the operation is simple, the consumption of fermentation and sterilization steam is low, the production water is less, the discharge of waste water generated by washing a fermentation tank of a fermentation tank is reduced, and the effects of energy conservation, consumption reduction and clean production are achieved.
The technical scheme adopted by the invention is as follows:
a preparation method of high-density yeast fermentation liquor is characterized by comprising the following steps:
①, preparing a dilute compound nutrient solution and a concentrated compound nutrient solution:
crushing corn and bean pulp, adding water for size mixing, sequentially adding high-temperature amylase, saccharifying enzyme and protease, performing temperature-controlled enzymolysis, and performing thorough enzymolysis to obtain liquid enzymolysis liquid; carrying out solid-liquid separation on the liquid enzymolysis liquid to prepare solid wet residues and a dilute composite nutrient solution;
reserving a part of the dilute composite nutrient solution as fermentation substrate sugar, and performing reduced pressure evaporation and concentration on the rest of the dilute composite nutrient solution to obtain a concentrated composite nutrient solution;
② preparation of high-density yeast liquid
In a full-automatic fermentation tank, diluting a part of dilute composite nutrient solution, sterilizing at high temperature, cooling, inoculating yeast seed solution, introducing sufficient sterile air for culture, controlling sufficient dissolved oxygen by respectively feeding the concentrated composite nutrient solution and diammonium hydrogen phosphate solution in a flowing manner in the culture process, stopping feeding when the yeast density is maximum, and preparing yeast fermentation liquor after the fermentable sugar is consumed;
keeping the fermentation tank at positive pressure, standing the yeast fermentation liquor, and discharging the precipitated yeast from the tank bottom after complete sedimentation to obtain high-density yeast liquid; and continuously adding the residual fermentation liquor in the fermentation tank with the concentrated composite nutrient solution and the diammonium hydrogen phosphate solution in a flowing manner to control sufficient dissolved oxygen, performing enrichment culture on the yeast again to the maximum density, standing, discharging high-density yeast liquid, continuously and circularly producing for multiple times, and collecting all the high-density yeast liquid.
Further, the yeast seed liquid is saccharomyces cerevisiae CICC 32280.
Further, the reaction conditions in step ① are as follows:
the mass ratio of the corn to the soybean meal is 1: 0.01-0.2; adding water to adjust the concentration of the dry matter to 10-40%; the addition amount of the high-temperature amylase is 0.1-1 kg per ton of the corn and soybean meal mixture, the pH value of the slurry is controlled to be 5.5-6.5, and the enzymolysis time is 0.5-2 hours;
the addition amount of the saccharifying enzyme is that 0.1-0.5 kg of controlled slurry pH 4.5-5.5 is added to each ton of corn and soybean meal mixture for enzymolysis for 0.5-20 h;
the addition amount of the protease is 0.1-1 kg per ton of the corn and soybean meal mixture, the pH of the slurry is controlled to be 4.5-5.5, and the enzymolysis time is 0.5-20 h;
the concentration of the concentrated dry matter of the dilute composite nutrient solution is 40-65%.
Preferably, the reaction conditions in step ① are as follows:
the mass ratio of the corn to the soybean meal is 1: 0.05-0.1; adding water to adjust the concentration of the dry matter to be 25-30%; the addition amount of the high-temperature amylase is 0.3-0.5 kg per ton of the corn and soybean meal mixture, the pH value of the slurry is controlled to be 5.8-6.2, and the enzymolysis time is 0.5-1 h;
the addition amount of the saccharifying enzyme is that 0.4-0.5 kg of controlled slurry pH4.8-5.2 is added to each ton of corn and soybean meal mixture for enzymolysis for 5-10 h;
the addition amount of the protease is that 0.5-0.6 kg of controlled slurry pH is added to each ton of corn and soybean meal mixture, the enzymolysis time is 15-20 h;
the concentration of the concentrated dry matter of the dilute composite nutrient solution is 40-45%.
Further, the reaction conditions in step ② are as follows:
diluting the dilute composite nutrient solution to a dry matter concentration of 0.01-5%; the mass ratio of the diammonium hydrogen phosphate to the water in the diammonium hydrogen phosphate solution is 10-50: 50-90; the feeding mass ratio of the concentrated compound nutrient solution to the diammonium hydrogen phosphate solution is 1: 0.001-0.1; the inoculation mass ratio of the yeast seed liquid is 0.2-20%;
when preparing the yeast fermentation liquor: fermentation temperature: 20-40 ℃; fermentation pH: 4.0 to 6.0; the stirring speed of aerobic fermentation is 20-1000 r/min; the ventilation ratio is 1: 0.5-3 v/v; the concentration of dissolved oxygen is 10-100%; and (3) fermentation period: 12 to 72 hours, the number of the bacteria at the end point of the yeast fermentation is 0.5 to 1.20 multiplied by 109cfu/ml；
The yeast culture end marker is: the content of fermentable sugar in the fermentation liquor is 0-0.1%, and the content of dissolved oxygen is 60-100%; standing and settling time is 1-20 h.
Preferably, the reaction conditions in step ② are as follows:
diluting the dilute composite nutrient solution to a dry matter concentration of 0.5-2%; the mass ratio of the diammonium hydrogen phosphate to the water in the diammonium hydrogen phosphate solution is 30-40: 60-70 parts of; the feeding mass ratio of the concentrated compound nutrient solution to the diammonium hydrogen phosphate solution is 1: 0.005-0.01; the inoculation mass ratio of the yeast seed liquid is 0.2-20%;
when preparing the yeast fermentation liquor: fermentation temperature: 30-35 ℃; fermentation pH: 4.0 to 4.5; the rotation speed of aerobic fermentation stirring is 100-150 r/min; the ventilation ratio is 1: 0.5-2 v/v; the concentration of dissolved oxygen is 20-40%; and (3) fermentation period: 48-56 h, the number of the bacteria at the end point of the yeast fermentation is 1.15-1.20 multiplied by 109cfu/ml；
The yeast culture end marker is: the content of fermentable sugar in the fermentation liquor is 0-0.05%, and the content of dissolved oxygen is 60-80%; standing and settling time is 4-6 h.
Further, in the step ①, the mass percentage of a part of the dilute composite nutrient solution in the dilute composite nutrient solution is 1-2%.
Further, the continuous cycle production times in the step ② are 8-10 times.
The invention has the beneficial effects that:
1. cheap corn and bean pulp are subjected to enzymolysis through amylase, glucoamylase and protease to form reducing sugar and organic nitrogen source mixed nutrient solution which is suitable for the growth of yeast in required proportion, the reducing sugar and organic nitrogen source required for the growth of yeast are provided, and in addition, diammonium hydrogen phosphate solution is added to effectively replace dipotassium hydrogen phosphate and ammonium sulfate in the traditional process, so that phosphorus source and inorganic ammonia nitrogen required for the growth of yeast can be met, the sulfate ion content of fermentation liquor can be reduced, and the production raw material cost is saved.
2. By means of continuous feeding and concentration of the composite nutrient solution and diammonium hydrogen phosphate solution, the method can effectively control the dissolved oxygen content of the fermentation liquor to be stable, provide reducing sugar, nitrogen source and phosphorus source for sufficient yeast growth, avoid substrate inhibition caused by one-time feeding and reduction of fermentation efficiency caused by oxygen deficiency, obtain the maximum increment speed and the maximum density of the yeast, and further, keep the yeast cells to settle after the fermentation is finished, the high-density yeast liquid is obtained by the bottom batch partial discharge mode, the rest fermentation liquid can be continuously fed and fermented without high-temperature sterilization and cooling inoculation again, the high-density yeast fermentation liquor obtained by the method not only has the yeast content increased to 18 percent, but also has the advantages of simple operation, small consumption of fermentation and sterilization steam, less production water, reduction of the discharge amount of waste water from tank washing of a fermentation tank, and achievement of the effects of energy conservation, consumption reduction and clean production.
Detailed Description
Example 1
A preparation method of high-density yeast fermentation liquor comprises the following specific operation methods:
(1) and preparing the dilute composite nutrient solution and the concentrated composite nutrient solution:
crushing 95kg of corn and 5kg of soybean meal, adding 200kg of water to prepare slurry, adjusting the pH value to 6.0, adding 0.04kg of high-temperature amylase, and carrying out enzymolysis at the high temperature of 90 ℃ for 1 hour; cooling to 55 deg.C, adjusting pH to 5.2, adding 0.04kg saccharifying enzyme, and performing enzymolysis for 5 hr; adjusting the pH value to 4.8, adding 0.06kg of protease, and keeping the temperature for enzymolysis for 15h to obtain liquid enzymolysis liquid;
filtering the liquid enzymolysis liquid to obtain 260kg of filtrate which is the dilute composite nutrient solution, and evaporating and concentrating the dilute composite nutrient solution under reduced pressure to obtain 163kg of concentrated composite nutrient solution with the solid content of 44%.
(2) The production method of the high-density yeast liquid comprises the following steps:
①, diluting 2kg of dilute composite nutrient solution to 85L in a 200L full-automatic fermentation tank, wherein the concentration of dry matters is 0.5%, the feed liquid is heated to 90 ℃, the tank body above the liquid level is heated to 121 ℃ for sterilization, the heat preservation sterilization time is 30 minutes, the temperature is reduced to 35 ℃, 5L of yeast seed solution is inoculated, the initial volume is observed to be 105L, the initial ventilation volume is adjusted to be 1:0.5v/v, the stirring speed is 100r/min, the rotating speed and the ventilation ratio are improved when the dissolved oxygen is lower than 20%, the culture is carried out for 8h, when the dissolved oxygen is suddenly increased to be more than 60%, the sterile concentrated composite nutrient solution and diammonium hydrogen phosphate solution are fed through a peristaltic pump, the feeding mass ratio of the concentrated composite nutrient solution and diammonium hydrogen phosphate is 1:0.005, the pH is controlled to be 4.0-4.5, the dissolved oxygen is controlled to be 20-40%, the stirring speed is increased to be 150r/min, the ventilation volume is 1:2v/v, the culture is carried out for 56h, the volume9cfu/ml, stopping feeding, and stopping ventilation and stirring until the dissolved oxygen rises to 70 percent. Standing for 4h to allow yeast to naturally settle, discharging 52L of the settled yeast through a discharge valve at the bottom of the tank, and detecting that the dry matter content of the yeast reaches 182 g/L.
②, closing a tank bottom valve, recovering the ventilation quantity to be 1:2v/v, stirring at the rotating speed of 150r/min, feeding sterile concentrated composite nutrient solution and diammonium phosphate solution again through a peristaltic pump, controlling the pH to be 4.0-4.5, controlling the dissolved oxygen to be 20% -40%, continuing fermentation culture for 52h, stopping feeding when the volume of fermentation liquor reaches 168L, stopping feeding until the dissolved oxygen rises back to 70%, stopping ventilation and stirring, standing for 6h, naturally settling the yeast, discharging 56L of precipitated yeast through a tank bottom discharge valve, and detecting that the content of dry matters in the yeast reaches 186 g/L.
Example 2
A preparation method of high-density yeast fermentation liquor comprises the following specific steps:
(1) and preparing the dilute composite nutrient solution and the concentrated composite nutrient solution:
pulverizing 1850kg of semen Maydis and 150kg of soybean meal, adding 4400kg of water to obtain slurry, adjusting pH to 6.0, adding 0.8kg of high temperature amylase, and performing enzymolysis at 90 deg.C for 1 hr; cooling to 55 deg.C, adjusting pH to 5.2, adding 0.8kg saccharifying enzyme, and performing enzymolysis for 5 hr; adjusting the pH value to 4.8, adding 1.2kg of protease, and keeping the temperature for enzymolysis for 15 h; 5416kg of filtrate is obtained by filtering, namely the dilute compound nutrient solution, and the concentrated compound nutrient solution 3520kg is obtained by carrying out reduced pressure evaporation and concentration on the dilute compound nutrient solution, wherein the solid content is 40%.
(2) The production method of the high-density yeast liquid comprises the following steps:
①, in a 2m3 fermentation tank, diluting 25kg of dilute composite nutrient solution to 0.9m3, wherein the concentration of dry matters is 0.6%, heating the feed liquid to 90 ℃, heating the tank above the liquid level to 121 ℃ for sterilization, keeping the temperature for sterilization for 30 minutes, cooling to 35 ℃, inoculating 100L of yeast seed liquid, observing and recording the initial volume to be 1.1m3, adjusting the initial ventilation amount to be 1:0.5v/v, stirring the rotation speed to be 100r/min, increasing the rotation speed and the ventilation ratio when the dissolved oxygen is lower than 20%, culturing for 6h, feeding sterile concentrated composite nutrient solution and diammonium hydrogen phosphate solution by a peristaltic pump when the dissolved oxygen is suddenly increased to be more than 60%, feeding the mass ratio of the concentrated composite nutrient solution to the diammonium hydrogen phosphate to be 1:0.004, controlling the pH to be 4.0-4.5, controlling the dissolved oxygen to be 20-40%, increasing the stirring rotation speed to be 150r/min, detecting the ventilation amount to be 1:1.8v/v, culturing for 56h, and detecting the volume of fermentation broth to be 1.54m3, and detecting the number of yeast9cfu/ml, stopping feeding, and stopping ventilation and stirring until the dissolved oxygen rises to 70 percent. Standing for 6 hr to allow yeast to settle naturally and openThe settled yeast is discharged by 0.44m3 through the discharge valve at the bottom of the tank, and the dry matter content of the yeast is detected to reach 182 g/L.
②, closing a tank bottom valve, recovering the ventilation quantity to be 1:1.8v/v, stirring at the rotating speed of 150r/min, feeding sterile concentrated composite nutrient solution and diammonium hydrogen phosphate solution by a peristaltic pump again, controlling the pH to be 4.0-4.5, controlling the dissolved oxygen to be 20% -40%, continuing fermentation culture for 50h, stopping feeding when the volume of fermentation liquor reaches 1.45m3, stopping feeding when the dissolved oxygen rises to 70%, stopping ventilation and stirring, standing for 6h, naturally settling the yeast, discharging the settled yeast by 0.44m3 through a tank bottom discharge valve, and detecting that the dry matter content of the yeast reaches 183 g/L.
③, closing a tank bottom valve, recovering the ventilation quantity to be 1:1.8v/v, stirring at the rotating speed of 150r/min, feeding sterile concentrated composite nutrient solution and diammonium hydrogen phosphate solution by a peristaltic pump again, controlling the pH to be 4.0-4.5, controlling the dissolved oxygen to be 20% -40%, continuing fermentation culture for 52h, stopping feeding when the volume of fermentation liquor reaches 1.46m3, stopping feeding when the dissolved oxygen rises back to 70%, stopping ventilation and stirring, standing for 6h, naturally settling the yeast, discharging the settled yeast by 0.46m3 through a tank bottom discharge valve, and detecting that the dry matter content of the yeast reaches 184 g/L.
④, 8 batches of the culture were repeated in this way, and the yeast still grew well and the yeast concentration reached 180g/L or more, and a yeast liquid with a high concentration of 3.67m3 was obtained in total.
Comparative example 1
Comparative example 1 is a conventional yeast preparation method, which specifically comprises:
① the fermentation medium comprises industrial glucose 65g/L, urea 1.8g/L, industrial corn flour 10g/L, KH2PO410g/L，MgSO44g/L，ZnSO410mg/L，FeSO412mg/L，MnSO42mg/L。
② feed medium is prepared, 500g/L glucose solution.
③ inoculating Saccharomyces cerevisiae seed liquid at 10% inoculation amount in 2m3 fermenter for fermentation at 30 deg.C, pH5.5, liquid loading 0.9m3, initial rotation speed 150r/min, initial aeration 1:2v/v, starting feeding at 2.4L/h when fermentation is 10h, stirring rotation speed is increased to 200r/min, adjusting aeration ratio to 1:2.5v/v, changing dissolved oxygen in middle and later stage of fermentation, if dissolved oxygen is suddenly increased to above 30%, slightly increasing feed supplement rate, controlling dissolved oxygen to maintain about 30%, ending fermentation period of 55h, dry weight of Saccharomyces cerevisiae cell of 122g/L, cell number of fermentation broth of 1.05 × 109cfu/ml。
The yeast cell number and dry matter concentration of fermentation broth of the first, example 1, example 2 and comparative example 1 are shown in table 1 below.
Table 1, example 2 and comparative example 1 fermentation broth yeast cell number and retort yeast dry matter concentration
As can be seen from Table 1, the fermentation mode of controlling dissolved oxygen by feeding and enzymolysis of the composite nutrient solution during fermentation can obtain 1.19X 10 cells at the end of fermentation9cfu/ml is improved by 13% compared with the traditional fermentation mode of example 1, the concentration of dry substances of high-concentration yeast liquid obtained by a standing precipitation batch discharging mode in the fermentation process reaches 186g/L, the supernatant can continue to adopt a feed supplement mode to ferment and culture yeast, the fermentation efficiency is improved by 52% compared with the traditional fermentation mode of example 1, the utilization rate of equipment is greatly improved, the operation is simple, the consumption of fermentation sterilization steam is small, the production water is less, the discharge amount of waste water in tank washing of a fermentation tank is reduced, and the effects of energy conservation, consumption reduction and clean production are achieved.
Claims (8)
1. A preparation method of high-density yeast fermentation liquor is characterized by comprising the following steps:
①, preparing a dilute compound nutrient solution and a concentrated compound nutrient solution:
crushing corn and bean pulp, adding water for size mixing, sequentially adding high-temperature amylase, saccharifying enzyme and protease, performing temperature-controlled enzymolysis, and performing thorough enzymolysis to obtain liquid enzymolysis liquid; carrying out solid-liquid separation on the liquid enzymolysis liquid to prepare solid wet residues and a dilute composite nutrient solution;
reserving a part of the dilute composite nutrient solution as fermentation substrate sugar, and performing reduced pressure evaporation and concentration on the rest of the dilute composite nutrient solution to obtain a concentrated composite nutrient solution;
② preparation of high-density yeast liquid
In a full-automatic fermentation tank, diluting a part of dilute composite nutrient solution, sterilizing at high temperature, cooling, inoculating yeast seed solution, introducing sufficient sterile air for culture, controlling sufficient dissolved oxygen by respectively feeding the concentrated composite nutrient solution and diammonium hydrogen phosphate solution in a flowing manner in the culture process, stopping feeding when the yeast density is maximum, and preparing yeast fermentation liquor after the fermentable sugar is consumed;
keeping the fermentation tank at positive pressure, standing the yeast fermentation liquor, and discharging the precipitated yeast from the tank bottom after complete sedimentation to obtain high-density yeast liquid; and continuously adding the residual fermentation liquor in the fermentation tank with the concentrated composite nutrient solution and the diammonium hydrogen phosphate solution in a flowing manner to control sufficient dissolved oxygen, performing enrichment culture on the yeast again to the maximum density, standing, discharging high-density yeast liquid, continuously and circularly producing for multiple times, and collecting all the high-density yeast liquid.
2. The method according to claim 1, wherein the method comprises the steps of: the yeast seed liquid is Saccharomyces cerevisiae CICC 32280.
3. The method of claim 1, wherein the reaction conditions in step ① are as follows:
the mass ratio of the corn to the soybean meal is 1: 0.01-0.2; adding water to adjust the concentration of the dry matter to 10-40%; the addition amount of the high-temperature amylase is 0.1-1 kg per ton of the corn and soybean meal mixture, the pH value of the slurry is controlled to be 5.5-6.5, and the enzymolysis time is 0.5-2 hours;
the addition amount of the saccharifying enzyme is that 0.1-0.5 kg of controlled slurry pH 4.5-5.5 is added to each ton of corn and soybean meal mixture for enzymolysis for 0.5-20 h;
the addition amount of the protease is 0.1-1 kg per ton of the corn and soybean meal mixture, the pH of the slurry is controlled to be 4.5-5.5, and the enzymolysis time is 0.5-20 h;
the concentration of the concentrated dry matter of the dilute composite nutrient solution is 40-65%.
4. The method of claim 3, wherein the reaction conditions in step ① are as follows:
the mass ratio of the corn to the soybean meal is 1: 0.05-0.1; adding water to adjust the concentration of the dry matter to be 25-30%; the addition amount of the high-temperature amylase is 0.3-0.5 kg per ton of the corn and soybean meal mixture, the pH value of the slurry is controlled to be 5.8-6.2, and the enzymolysis time is 0.5-1 h;
the addition amount of the saccharifying enzyme is that 0.4-0.5 kg of controlled slurry pH4.8-5.2 is added to each ton of corn and soybean meal mixture for enzymolysis for 5-10 h;
the addition amount of the protease is that 0.5-0.6 kg of controlled slurry pH is added to each ton of corn and soybean meal mixture, the enzymolysis time is 15-20 h;
the concentration of the concentrated dry matter of the dilute composite nutrient solution is 40-45%.
5. The method of claim 1, wherein the reaction conditions in step ② are as follows:
diluting the dilute composite nutrient solution to a dry matter concentration of 0.01-5%; the mass ratio of the diammonium hydrogen phosphate to the water in the diammonium hydrogen phosphate solution is 10-50: 50-90; the feeding mass ratio of the concentrated compound nutrient solution to the diammonium hydrogen phosphate solution is 1: 0.001-0.1; the inoculation mass ratio of the yeast seed liquid is 0.2-20%;
when preparing the yeast fermentation liquor: fermentation temperature: 20-40 ℃; fermentation pH: 4.0 to 6.0; the stirring speed of aerobic fermentation is 20-1000 r/min; the ventilation ratio is 1: 0.5-3 v/v; the concentration of dissolved oxygen is 10-100%; and (3) fermentation period: 12 to 72 hours, the number of the bacteria at the end point of the yeast fermentation is 0.5 to 1.20 multiplied by 109cfu/ml；
The yeast culture end marker is: the content of fermentable sugar in the fermentation liquor is 0-0.1%, and the content of dissolved oxygen is 60-100%; standing and settling time is 1-20 h.
6. The method of claim 5, wherein the reaction conditions in step ② are as follows:
diluting the dilute composite nutrient solution to a dry matter concentration of 0.5-2%; the mass ratio of the diammonium hydrogen phosphate to the water in the diammonium hydrogen phosphate solution is 30-40: 60-70 parts of; the feeding mass ratio of the concentrated compound nutrient solution to the diammonium hydrogen phosphate solution is 1: 0.005-0.01; the inoculation mass ratio of the yeast seed liquid is 0.2-20%;
when preparing the yeast fermentation liquor: fermentation temperature: 30-35 ℃; fermentation pH: 4.0 to 4.5; the rotation speed of aerobic fermentation stirring is 100-150 r/min; the ventilation ratio is 1: 0.5-2 v/v; the concentration of dissolved oxygen is 20-40%; and (3) fermentation period: 48-56 h, the number of the bacteria at the end point of the yeast fermentation is 1.15-1.20 multiplied by 109cfu/ml；
The yeast culture end marker is: the content of fermentable sugar in the fermentation liquor is 0-0.05%, and the content of dissolved oxygen is 60-80%; standing and settling time is 4-6 h.
7. The method for preparing high-density yeast fermentation broth according to claim 1, wherein the mass percentage of a part of the dilute composite nutrient solution in step ① in the dilute composite nutrient solution is 1-2%.
8. The method according to claim 1, wherein the number of continuous cycles in step ② is 8-10.
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| CN113462631A (en) * | 2021-06-30 | 2021-10-01 | 金华职业技术学院 | Strain fermentation process beneficial to improving strain density |
| CN113652361A (en) * | 2021-01-29 | 2021-11-16 | 上海源耀农牧科技有限公司 | High-density fermentation process of yellow wine yeast |
| CN116676203A (en) * | 2023-06-08 | 2023-09-01 | 珠海盈盛联科生物科技有限公司 | Preparation process of amino acid cereal yeast applied to skin care product |
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