CN112143661A - Method for producing microbial peptone by using vegetable oil as raw material - Google Patents
Method for producing microbial peptone by using vegetable oil as raw material Download PDFInfo
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Abstract
The invention discloses a method for producing microbial peptone by using vegetable oil as a raw material, which specifically comprises the following steps: 1. inoculating yeast seed liquid into a sterile fermentation medium containing vegetable oil, fermenting under aerobic conditions, and feeding the sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquid and supplementing nitrogen sources and trace nutrient substances into the fermentation liquid during fermentation to obtain fermentation liquid containing single-cell protein; 2. sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone. The method replaces the traditional carbon source in the fermentation medium with the vegetable oil, not only reduces the cost of raw materials, but also can utilize the vegetable oil and develop the application field of the vegetable oil; in addition, the conversion rate of yeast microorganisms is improved, the single-cell peptone with high added value is produced, the production cost is greatly reduced, and the production process is simplified.
Description
Technical Field
The invention relates to the technical field of biological fermentation, in particular to a method for producing microbial peptone by using vegetable oil as a raw material.
Background
Peptone is a soluble mixture obtained by partial enzymolysis or acid hydrolysis of proteins from different sources. Peptone is rich in organic nitrogen compounds and also contains some vitamins and sugars. It can be used as main raw material of microbial culture medium, and can be extensively used in the fields of antibiotic, medicine industry, fermentation industry, biochemical product and microbiological research, etc.
Peptone can be classified from a source into animal peptone, plant peptone, and microbial (yeast) peptone.
Animal and plant peptone is the major peptone in the market, but the growth state of animals and plants and the content of protein in the animals and plants can fluctuate due to the growth environment of the animals and plants and some external factors, and the storage, extraction and processing system of the protein is basically open, so that the quality of raw materials can change (such as discoloration, decay and the like) and the difference of nutrient components can be caused, and finally the quality of peptone products is unstable. In addition, animal peptone may have pathogenicity and colloquial contraindication problems, and vegetable peptone mainly has anaphylaxis and transgenic disputed problems. The protein raw material of the microorganism (yeast) peptone is derived from yeast, has no problems of transgenic steam, pathogenicity, allergic source, popular contraindication and the like, and is favorable for passing international certification of KOSHER, HALAL and the like.
In addition, from the nutrition aspect, the animal and plant source peptone has single nutrient component, and the microorganism (yeast) peptone is rich in protein, peptides, amino acid, nucleotide, B vitamins, biotin and the like, and can provide comprehensive and balanced nutrition for thalli. The combined use of yeast extract and other media components can promote the growth and metabolism of microorganisms, provide production efficiency, and thus the market demand of microbial (yeast) peptone is becoming more and more extensive.
Peptone is a soluble mixture obtained by partial enzymolysis or acid hydrolysis of proteins from different sources. However, in the production cost of protein (single cell protein), the raw material cost still accounts for more than 60% of the total production cost, wherein the cost of the carbon source in the raw material is the highest, and the yield of the single cell protein is not ideal, so that how to improve the conversion rate of microorganisms such as yeast and the like and produce the single cell peptone with high additional value is a difficult problem which is difficult to overcome in the field.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: the method for producing the microbial peptone by using the vegetable oil as the raw material is provided, the vegetable oil is used for replacing a conventionally used carbon source, so that the raw material cost is reduced, the vegetable oil can be used, and the application field of the vegetable oil is developed; in addition, the method optimizes the strains, the fermentation process and the culture medium, improves the conversion rate of yeast microorganisms, produces the single-cell peptone with high added value, greatly reduces the production cost and simplifies the production process.
In order to solve the problems, the method for producing the microbial peptone by using the vegetable oil as the raw material specifically comprises the following steps:
the method comprises the following steps: inoculating yeast seed liquid into a sterile fermentation medium containing vegetable oil, fermenting under aerobic conditions, and finally fermenting to obtain a fermentation liquid containing single-cell protein; feeding sterile vegetable oil and ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source into the fermentation liquor during the fermentation period, or feeding sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquor and supplementing the nitrogen source and micronutrients into the fermentation liquor during the fermentation period;
step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery microbial peptone.
Wherein the sterile vegetable oil is obtainable by autoclaving a vegetable oil. The vegetable oil can adopt one or more of the following (1), (2), (3) and (4) to mix:
(1) industrial vegetable oils and edible vegetable oils;
(2) vegetable oil with high acid value or not meeting the national quality standard;
(3) vegetable oil used for waste after frying food;
(4) distillate produced in the processes of deacidification and deodorization of the vegetable oil production process.
The aerobic condition refers to the existence of oxygen in the microbial fermentation environment, and can be obtained by the conventional method in the field, for example, a certain dissolved oxygen can be ensured by setting the stirring rotating speed and the ventilation quantity in the fermentation process. In the fermentation period, the dissolved oxygen in the fermentation liquor is 5 to 50 percent by the cooperation of aeration quantity and fermentation liquor stirring.
In the present invention, the yeast seed solution can be obtained by a method conventional in the art, and is usually obtained by inoculating yeast into a seed culture medium for fermentation. The seed medium is chosen by the person skilled in the art as a function of the type of yeast to be inoculated, and may be, for example, a glucose yeast peptone medium. The fermentation conditions for the seed liquid may be those conventional in the art suitable for seed liquids. In the first step, the yeast in the yeast seed liquid is: one or more of candida utilis, candida tropicalis, yarrowia lipolytica and oleaginous yeast. The content of the vegetable oil in the fermentation medium containing the vegetable oil is 5-20 g/L, and preferably 8-15 g/L. The inoculation concentration of the yeast seed liquid into the fermentation culture medium is 1-10%, preferably 3-5% of the volume of the fermentation culture medium.
The carbon source used in the fermentation medium of the invention is only vegetable oil. Usually, the seed liquid contains a certain carbon source, so when the seed liquid is inoculated into the fermentation medium, the fermentation liquid contains a very low concentration of carbon source, but the carbon source is rapidly consumed as the fermentation proceeds. However, as the initial fermentation medium, i.e., the fermentation medium before inoculation, the conventional fermentation carbon source is replaced with vegetable oil, and therefore, the fermentation medium does not contain any carbon source other than vegetable oil. In addition to replacing the carbon source with vegetable oil, other ingredients in the fermentation medium of the present invention are conventional ingredients used in microbial fermentation. In other words, the fermentation medium of the present invention can be regarded as a medium modified on the basis of a conventional fermentation medium in such a manner that all carbon sources in the fermentation medium are replaced with vegetable oils, and the remaining components are the same as those of the conventional fermentation medium. Therefore, the components of the fermentation medium used in the present invention, other than the vegetable oil, are components conventionally used in the field for fermentation of various microorganisms, but the concentration of each component in the medium is optimized according to the characteristics of yeast, unlike the conventional medium. The fermentation medium comprises the following components: 0.5-2.5 g/L of yeast extract, 1-3 g/L of magnesium sulfate, 5-20 g/L of nitrogen source, 2-5 g/L of monopotassium phosphate and/or dipotassium phosphate and one or more of trace elements; the trace elements comprise one or more components, and the content of each component is 0.002-0.08 g/L.
When the fermentation medium comprises a nitrogen source, the nitrogen source is ammonium sulfate.
When the components of the fermentation medium contain trace elements, the components of the trace elements specifically include: iron chloride: 0.01-0.02 g/L, manganese sulfate: 0.01-0.02 g/L, zinc sulfate: 0.01-0.02 g/L, copper sulfate: 2-4 mg/L, sodium molybdate: 2-4 mg/L, calcium chloride: 0.02-0.08 g/L, sodium iodide: 2-5 mg/L, folic acid: 2-8 mg/L, citric acid: 10-40 mg/L, vitamin C: 2-12 mg/L, vitamin D: 2-12 mg/L, vitamin E: 2-12 mg/L, vitamin B6: 2-20 mg/L, vitamin B12: 2-20 mg/L.
Further, in the method for producing microbial peptone by using vegetable oil as a raw material, in the first step, when micronutrients are fed into the fermentation broth during fermentation, the trace elements in the micronutrients are as follows: one or more of sodium, phosphorus, potassium, magnesium, calcium, iron, copper, zinc, molybdenum, iodine, folic acid, citric acid, vitamin C, vitamin D, vitamin E, vitamin B6, and vitamin B12.
Further, in the method for producing microbial peptone using vegetable oil as a raw material, the components of the micronutrients and the contents of the components in the fermentation broth after the components are added to the fermentation broth are as follows: magnesium sulfate: 1-3 g/L, monopotassium phosphate and/or dipotassium phosphate: 2-5 g/L, ferric chloride: 0.01-0.02 g/L, manganese sulfate: 0.01-0.02 g/L, zinc sulfate: 0.01-0.02 g/L, copper sulfate: 2-4 mg/L, sodium molybdate: 2-4 mg/L, calcium chloride: 0.02-0.08 g/L, sodium iodide: 2-5 mg/L, folic acid: 2-8 mg/L, citric acid: 10-40 mg/L, vitamin C: 2-12 mg/L, vitamin D: 2-12 mg/L, vitamin E: 2-12 mg/L, vitamin B6: 2-20 mg/L, vitamin B12: 2-20 mg/L.
Further, in the method for producing microbial peptone by using vegetable oil as a raw material, the fermentation temperature of the fermentation liquid during the fermentation is 25-35 ℃, preferably 29-31 ℃. The ventilation amount during the fermentation is 0.5-2.5 vvm; the ventilation amount during the fermentation period can be one of three ranges of 1-1.5 vvm, 1.5-2 vvm and 1.8-2.5 vvm. During fermentation, the dissolved oxygen in the fermentation liquor is 5 to 50 percent through the coordination of ventilation and fermentation liquor stirring; adding ammonia water into the fermentation liquor during fermentation to maintain the pH value of the fermentation liquor at 6.2-8, preferably 7.2-7.8; the ammonia water is selected with the concentration of 10-20%. And (3) feeding sterile vegetable oil into the fermentation liquor during the fermentation period so as to maintain the volume ratio of the total fat content in the fermentation liquor at 0.5-2%, preferably at one of the ranges of 0.5-1.5% and 1-2%.
Further, in the method for producing microbial peptone by using vegetable oil as a raw material, in the second step, when the fermentation broth containing the single-cell protein is subjected to precipitation separation and centrifugation in sequence, the centrifugation rotation speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes, so that the concentration of the single-cell protein in the fermentation broth containing the single-cell protein is about 20%.
When a high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150MPa, and the cell disruption rate under a microscope reaches more than 90 percent.
When the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃. Finally obtaining the powdered microbial peptone with the water content of less than 8%.
Further, in the method for producing microbial peptone by using vegetable oil as a raw material, in the second step, the enzymolysis process is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
The invention has the beneficial effects that: the method replaces the traditional carbon source in the fermentation culture medium with the vegetable oil, not only reduces the cost of raw materials, but also can utilize the vegetable oil and develop the application field of the vegetable oil; secondly, the strain, fermentation process and culture medium are optimized, the conversion rate of yeast microorganisms is improved, the single-cell peptone with high added value is produced, the production cost is greatly reduced, and the production process is simplified.
Detailed Description
The method for producing the microbial peptone by using the vegetable oil as the raw material can be applied to batch fermentation occasions and also can be applied to continuous fermentation occasions. The technical solution of the present invention will be further described in detail with reference to the preferred embodiments.
Example one
This example illustrates the batch fermentation production of microbial peptone in a 5L fermentor.
The method for producing microbial peptone by using vegetable oil as a raw material specifically comprises the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of fermentation medium containing sterile vegetable oil and sterilized by autoclaving at 121 ℃ is put into a 5L fermentation tank, the autoclaving time is 30 minutes, the pH value of the fermentation medium is 7.5, and the vegetable oil content in the fermentation medium containing vegetable oil is 5-20 g/L. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And (3) feeding sterile vegetable oil and ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source into the fermentation liquor during fermentation to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 5g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: 0.01g/L of ferric chloride, 0.02g/L of manganese sulfate, 0.02g/L of zinc sulfate, 2mg/L of copper sulfate and 2mg/L of sodium molybdate.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 15% concentrated ammonia water for automatic feeding regulation. And (2) initially feeding sterile vegetable oil into the fermentation tank according to the proportion of 10g/L, supplementing the sterile vegetable oil into the fermentation liquor when the total fatty acid content in the fermentation liquor is lower than 0.5%, and adjusting the total fatty acid content in the fermentation liquor by adding the sterile vegetable oil to maintain the total fatty acid content in the fermentation liquor at 0.5-2%.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying treatment on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
In the fermentation process, the mass conversion rate of the vegetable oil peptone is more than or equal to 45.1 percent.
When the vegetable oil is soybean oil, the total amount of the soybean oil is 46g/kg, the weight of the peptone is 23.8g/kg, and the mass conversion rate of the peptone prepared from the soybean oil is 51.7 percent.
When the vegetable oil is selected as palm oil, the total amount of palm oil used is 42g/kg, the weight of peptone obtained is 20.29g/kg, and the mass conversion rate of palm oil peptone is 48.3%.
When peanut oil is selected as the vegetable oil, 39g/kg of peanut oil is used together, the weight of peptone is 17.6g/kg, and the mass conversion rate of the peanut oil peptone is 45.1%.
When vegetable oil is selected from rapeseed oil, the total use amount of the rapeseed oil is 48g/kg, the weight of the peptone is 23.62g/kg, and the mass conversion rate of the rapeseed oil peptone is 49.2%.
Comparative example 1
And (3) taking a common carbon source on the market as a fermentation raw material to perform fermentation culture comparison, wherein the carbon source adopts glucose. The method for producing the microbial peptone by using glucose as a raw material specifically comprises the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of a fermentation medium containing sterile glucose autoclaved at 121 ℃ were placed in a 5L fermenter, and the autoclaving time was 30 minutes and the pH of the fermentation medium was 7.5. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And during fermentation, feeding sterile glucose solution, ammonia water for regulating the pH value of the fermentation liquor and supplementing a nitrogen source into the fermentation liquor to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 5g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: 0.01g/L of ferric chloride, 0.02g/L of manganese sulfate, 0.02g/L of zinc sulfate, 2mg/L of copper sulfate and 2mg/L of sodium molybdate.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 15% concentrated ammonia water for automatic feeding regulation. And initially feeding a sterile glucose solution with the concentration of 40% into the fermentation tank according to 10g/L, and when the concentration of the glucose in the fermentation liquor is lower than 5g/L, feeding the sterile glucose solution into the fermentation liquor to control the concentration of the glucose in the fermentation liquor to be 5-20 g/L.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
During this fermentation, a total of 66.3g/kg glucose was used, giving a peptone weight of 18.6g/kg and a glucose-made peptone mass conversion of 28.1%.
From the above first example and the first comparative example, it can be seen that: according to the invention, the vegetable oil is used for replacing the traditional carbon source in the fermentation medium, so that the raw material cost is reduced, and the vegetable oil can be utilized to develop the application field of the vegetable oil; in addition, the method optimizes the strains, the fermentation process and the culture medium, improves the conversion rate of yeast microorganisms, produces the single-cell peptone with high added value, greatly reduces the production cost and simplifies the production process.
Comparative example 2
This example illustrates the production of microbial peptone by automatic feeding of 25% strength ammonia water as a nitrogen source in a 5L fermenter.
The method for producing the microbial peptone by using the vegetable oil as the raw material specifically comprises the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of fermentation medium containing vegetable oil and sterilized at 121 ℃ under high pressure is put into a 5L fermentation tank, the autoclaving time is 30 minutes, the pH value of the fermentation medium is 7.5, and the vegetable oil content in the fermentation medium containing vegetable oil is 5-20 g/L. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And (3) feeding sterile vegetable oil and ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source into the fermentation liquor during fermentation to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 5g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: 0.01g/L of ferric chloride, 0.02g/L of manganese sulfate, 0.02g/L of zinc sulfate, 2mg/L of copper sulfate and 2mg/L of sodium molybdate.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 25% concentrated ammonia water for automatic feeding regulation. The automatic feeding regulation and control equipment is the same as that in the first embodiment, so that the volume of each part of concentrated ammonia water fed in a feeding way is the same, and the higher the concentration of the concentrated ammonia water is, the higher the nitrogen source content of each part of concentrated ammonia water fed in the fermentation liquor is.
And (2) initially feeding sterile vegetable oil into the fermentation tank according to the proportion of 10g/L, supplementing the sterile vegetable oil into the fermentation liquor when the total fatty acid content in the fermentation liquor is lower than 0.5%, and adjusting the total fatty acid content in the fermentation liquor by adding the sterile vegetable oil to maintain the total fatty acid content in the fermentation liquor at 0.5-2%.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
When soybean oil is selected as the vegetable oil in the fermentation process, 39g/kg of the soybean oil is used together, the weight of the peptone is 17.6g/kg, and the mass conversion rate of the peptone prepared from the soybean oil is 45.1%.
From the above first and second comparative examples: under the condition that the strains, the fermentation process, the using equipment and the culture medium are the same, the concentrated ammonia water with the use concentration of 25% is fed into the fermentation liquor during the fermentation period, the mass conversion rate of the finally obtained vegetable oil peptone is lower, the concentrated ammonia water with the use concentration of 10% -20% is fed into the fermentation liquor during the fermentation period, and the mass conversion rate of the finally obtained vegetable oil peptone is relatively higher.
Example two
This example differs from the first example in the selection of trace elements in the fermentation medium.
In this embodiment, the method for producing microbial peptone from vegetable oil as a raw material specifically includes the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of fermentation medium containing vegetable oil and sterilized at 121 ℃ under high pressure is put into a 5L fermentation tank, the autoclaving time is 30 minutes, the pH value of the fermentation medium is 7.5, and the vegetable oil content in the fermentation medium containing vegetable oil is 5-20 g/L. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And during fermentation, feeding sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source and trace nutrient substances into the fermentation liquor to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 5g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: iron chloride: 0.01g/L, manganese sulfate: 0.01g/L, zinc sulfate: 0.01g/L, copper sulfate: 3mg/L, sodium molybdate: 3mg/L, calcium chloride: 0.08g/L, sodium iodide: 3mg/L, folic acid: 4mg/L, citric acid: 20mg/L, vitamin C: 6mg/L, vitamin D: 6mg/L, vitamin E: 6mg/L, vitamin B6: 10mg/L, vitamin B12: 10 mg/L.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 15% concentrated ammonia water for automatic feeding regulation. And (2) initially feeding sterile vegetable oil into the fermentation tank according to the proportion of 10g/L, supplementing the sterile vegetable oil into the fermentation liquor when the total fatty acid content in the fermentation liquor is lower than 0.5%, and adjusting the total fatty acid content in the fermentation liquor by adding the sterile vegetable oil to maintain the total fatty acid content in the fermentation liquor at 0.5-2%.
The composition components of the micronutrients fed-batch to the fermentation liquor during the fermentation period and the content of each composition component in the fermentation liquor after being added into the fermentation liquor are as follows: 1g/L of magnesium sulfate, 5g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate, and ferric chloride: 0.01g/L, manganese sulfate: 0.01g/L, zinc sulfate: 0.01g/L, copper sulfate: 3mg/L, sodium molybdate: 3mg/L, calcium chloride: 0.08g/L, sodium iodide: 3mg/L, folic acid: 4mg/L, citric acid: 20mg/L, vitamin C: 6mg/L, vitamin D: 6mg/L, vitamin E: 6mg/L, vitamin B6: 10mg/L, vitamin B12: 10 mg/L.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution for a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction, the pH value of the second reaction solution is maintained at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
When soybean oil is selected as the vegetable oil in the fermentation process, the total amount of the soybean oil used is 64g/kg, the weight of the obtained peptone is 49.3g/kg, and the mass conversion rate of the peptone prepared from the vegetable oil is up to 77%.
EXAMPLE III
This example differs from the second example in the selection of the amount of the minor constituents in the fermentation medium.
In this embodiment, the method for producing microbial peptone from vegetable oil as a raw material specifically includes the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of fermentation medium containing vegetable oil and autoclaved at 121 ℃ is put into a 5L fermentation tank, the autoclaving time is 30 minutes, the pH value of the fermentation medium is 7.5, and the vegetable oil content in the fermentation medium containing vegetable oil is 5-20 g/L. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And during fermentation, feeding sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source and trace nutrient substances into the fermentation liquor to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 10g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: iron chloride: 0.02g/L, manganese sulfate: 0.02g/L, zinc sulfate: 0.02g/L, copper sulfate: 4mg/L, sodium molybdate: 4mg/L, calcium chloride: 0.08g/L, sodium iodide: 5 mg/L, folic acid: 8mg/L, citric acid: 40mg/L, vitamin C: 12mg/L, vitamin D: 12mg/L, vitamin E: 12mg/L, vitamin B6: 20mg/L, vitamin B12: 20 mg/L.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 15% concentrated ammonia water for automatic feeding regulation. And (2) initially feeding sterile vegetable oil into the fermentation tank according to the proportion of 10g/L, supplementing the sterile vegetable oil into the fermentation liquor when the total fatty acid content in the fermentation liquor is lower than 0.5%, and adjusting the total fatty acid content in the fermentation liquor by adding the sterile vegetable oil to maintain the total fatty acid content in the fermentation liquor at 0.5-2%.
The composition components of the micronutrients fed-batch to the fermentation liquor during the fermentation period and the content of each composition component in the fermentation liquor after being added into the fermentation liquor are as follows: 1g/L magnesium sulfate, 10g/L ammonium sulfate, 3g/L potassium dihydrogen phosphate and/or dipotassium hydrogen phosphate, and trace elements: iron chloride: 0.02g/L, manganese sulfate: 0.02g/L, zinc sulfate: 0.02g/L, copper sulfate: 4mg/L, sodium molybdate: 4mg/L, calcium chloride: 0.08g/L, sodium iodide: 5 mg/L, folic acid: 8mg/L, citric acid: 40mg/L, vitamin C: 12mg/L, vitamin D: 12mg/L, vitamin E: 12mg/L, vitamin B6: 20mg/L, vitamin B12: 20 mg/L.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution for a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction, the pH value of the second reaction solution is maintained at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
When soybean oil is selected as the vegetable oil in the fermentation process, the total amount of the soybean oil used is 48g/kg, the weight of the obtained peptone is 29.9g/kg, and the mass conversion rate of the peptone prepared from the vegetable oil is as high as 62.3 percent.
Example four
This example differs from the first example in the selection of the amount of the minor elements in the fermentation medium.
In this embodiment, the method for producing microbial peptone from vegetable oil as a raw material specifically includes the following steps:
the method comprises the following steps: first, Candida utilis was inoculated into 100ml of a seed medium and fermented to prepare a yeast seed solution. The seed culture medium is glucose yeast peptone culture medium, and the components of the culture medium generally comprise: 20% of glucose, 10% of yeast extract and 20% of peptone. The fermentation process can be carried out in a shaking table, the rotating speed of the shaking table is usually 100-200 rpm, and the fermentation temperature of the yeast seed liquid is controlled at 29 ℃.
2L of fermentation medium containing vegetable oil and autoclaved at 121 ℃ is put into a 5L fermentation tank, the autoclaving time is 30 minutes, the pH value of the fermentation medium is 7.5, and the vegetable oil content in the fermentation medium containing vegetable oil is 5-20 g/L. After obtaining the yeast seed liquid, inoculating the yeast seed liquid into a 5L fermentation tank according to the inoculation concentration of 5%, and carrying out fermentation culture under aerobic conditions, wherein the fermentation culture period is 72 hours. And during fermentation, feeding sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source and trace nutrient substances into the fermentation liquor to obtain the fermentation liquor containing the single-cell protein.
Wherein the temperature of the fermentation liquor in the fermentation tank is controlled at 29 ℃, the initial stirring rotating speed of stirring equipment in the fermentation tank is 200rpm, the initial ventilation in the fermentation tank is 1vvm, and the stirring rotating speed and the ventilation are set to ensure that the dissolved oxygen in the fermentation liquor is 5-50% in the whole fermentation culture process.
The fermentation medium in this example comprises the following components: 1g/L of yeast extract, 1g/L of magnesium sulfate, 20g/L of ammonium sulfate, 3g/L of monopotassium phosphate and/or dipotassium phosphate and trace elements: iron chloride: 0.01g/L, manganese sulfate: 0.01g/L, zinc sulfate: 0.01g/L, copper sulfate: 2mg/L, sodium molybdate: 2mg/L, calcium chloride: 0.02g/L, sodium iodide: 3mg/L, folic acid: 2mg/L, citric acid: 10mg/L, vitamin C: 2mg/L, vitamin D: 2mg/L, vitamin E: 2mg/L, vitamin B6: 2mg/L, vitamin B12: 2 mg/L.
After the fermentation culture is started, ammonia water is fed into the fermentation liquor, and the pH value of the fermentation liquor is kept at 7.5 by using 15% concentrated ammonia water for automatic feeding regulation. And (2) initially feeding sterile vegetable oil into the fermentation tank according to the proportion of 10g/L, supplementing the sterile vegetable oil into the fermentation liquor when the total fatty acid content in the fermentation liquor is lower than 0.5%, and adjusting the total fatty acid content in the fermentation liquor by adding the sterile vegetable oil to maintain the total fatty acid content in the fermentation liquor at 0.5-2%.
The composition components of the micronutrients fed-batch to the fermentation liquor during the fermentation period and the content of each composition component in the fermentation liquor after being added into the fermentation liquor are as follows: 1g/L magnesium sulfate, 20g/L ammonium sulfate, 3g/L potassium dihydrogen phosphate and/or dipotassium hydrogen phosphate, and trace elements: iron chloride: 0.01g/L, manganese sulfate: 0.01g/L, zinc sulfate: 0.01g/L, copper sulfate: 2mg/L, sodium molybdate: 2mg/L, calcium chloride: 0.02g/L, sodium iodide: 3mg/L, folic acid: 2mg/L, citric acid: 10mg/L, vitamin C: 2mg/L, vitamin D: 2mg/L, vitamin E: 2mg/L, vitamin B6: 2mg/L, vitamin B12: 2 mg/L.
Step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery peptone with the molecular weight of 180-3500 Da.
Wherein, when the fermentation liquor containing the single cell protein is sequentially subjected to precipitation separation and centrifugation, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes.
Wherein, when the high-pressure homogenizer is used for cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa.
Wherein, the specific process of enzymolysis is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution for a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction, the pH value of the second reaction solution is maintained at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
Wherein, when the spray drying treatment is adopted, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
When soybean oil is selected as the vegetable oil in the fermentation process, the total amount of the soybean oil used is 56g/kg, the weight of the obtained peptone is 38.1g/kg, and the mass conversion rate of the peptone prepared from the vegetable oil is as high as 68.0 percent.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, but any modifications or equivalent variations made in accordance with the technical spirit of the present invention are within the scope of the present invention as claimed.
The invention has the advantages that: the method replaces the traditional carbon source in the fermentation culture medium with the vegetable oil, not only reduces the cost of raw materials, but also can utilize the vegetable oil and develop the application field of the vegetable oil; secondly, the strain, fermentation process and culture medium are optimized, the conversion rate of yeast microorganisms is improved, the single-cell peptone with high added value is produced, the production cost is greatly reduced, and the production process is simplified.
Claims (10)
1. The method for producing the microbial peptone by using the vegetable oil as the raw material is characterized by comprising the following steps: the method specifically comprises the following steps:
the method comprises the following steps: inoculating yeast seed liquid into a sterile fermentation medium containing vegetable oil, and fermenting under an aerobic condition to obtain a fermentation liquid containing single-cell protein; feeding sterile vegetable oil and ammonia water for adjusting the pH value of the fermentation liquor and supplementing a nitrogen source into the fermentation liquor during the fermentation period, or feeding sterile vegetable oil, ammonia water for adjusting the pH value of the fermentation liquor and supplementing the nitrogen source and micronutrients into the fermentation liquor during the fermentation period;
step two: sequentially carrying out precipitation separation, centrifugation, cell crushing by a high-pressure homogenizer, enzymolysis, filtration, ion exchange electrodialysis, ultrafiltration and spray drying on the fermentation liquor containing the single-cell protein to finally obtain the powdery microbial peptone.
2. The process for the production of microbial peptones from vegetable oils as claimed in claim 1, characterized in that: in the first step, the yeast in the yeast seed liquid is: one or more of candida utilis, candida tropicalis, yarrowia lipolytica and oleaginous ester producing yeast; the vegetable oil content in the fermentation medium containing the vegetable oil is 5-20 g/L; the inoculation concentration of the yeast seed liquid into the fermentation culture medium is 1-10% of the volume of the fermentation culture medium; the fermentation medium comprises the following components: 0.5-2.5 g/L of yeast extract, 1-3 g/L of magnesium sulfate, 5-20 g/L of nitrogen source, 2-5 g/L of monopotassium phosphate and/or dipotassium phosphate and one or more of trace elements; the trace elements comprise one or more components, and the content of each component is 0.002-0.08 g/L.
3. The process for the production of microbial peptones from vegetable oils as claimed in claim 2, characterized in that: in the first step, the content of the vegetable oil in the fermentation medium containing the vegetable oil is 8-15 g/L; the inoculation concentration of the yeast seed liquid into the fermentation culture medium is 3-5% of the volume of the fermentation culture medium; when the fermentation medium comprises a nitrogen source, the nitrogen source is ammonium sulfate; when the components of the fermentation medium contain trace elements, the trace elements comprise the following components: iron chloride: 0.01-0.02 g/L, manganese sulfate: 0.01-0.02 g/L, zinc sulfate: 0.01-0.02 g/L, copper sulfate: 2-4 mg/L, sodium molybdate: 2-4 mg/L, calcium chloride: 0.02-0.08 g/L, sodium iodide: 2-5 mg/L, folic acid: 2-8 mg/L, citric acid: 10-40 mg/L, vitamin C: 2-12 mg/L, vitamin D: 2-12 mg/L, vitamin E: 2-12 mg/L, vitamin B6: 2-20 mg/L, vitamin B12: 2-20 mg/L.
4. The process for the production of microbial peptones starting from vegetable oils according to claim 1 or 2 or 3, characterized in that: in the first step, when the trace nutrient substances are fed into the fermentation liquor during the fermentation, the trace elements in the trace nutrient substances are as follows: one or more of sodium, phosphorus, potassium, magnesium, calcium, iron, copper, zinc, molybdenum, iodine, folic acid, citric acid, vitamin C, vitamin D, vitamin E, vitamin B6, and vitamin B12.
5. The process for the production of microbial peptones from vegetable oils as claimed in claim 4, characterized in that: in the first step, the components of the micronutrients and the content of each component in the fermentation broth after the components are added into the fermentation broth are as follows: magnesium sulfate: 1-3 g/L, monopotassium phosphate and/or dipotassium phosphate: 2-5 g/L, ferric chloride: 0.01-0.02 g/L, manganese sulfate: 0.01-0.02 g/L, zinc sulfate: 0.01-0.02 g/L, copper sulfate: 2-4 mg/L, sodium molybdate: 2-4 mg/L, calcium chloride: 0.02-0.08 g/L, sodium iodide: 2-5 mg/L, folic acid: 2-8 mg/L, citric acid: 10-40 mg/L, vitamin C: 2-12 mg/L, vitamin D: 2-12 mg/L, vitamin E: 2-12 mg/L, vitamin B6: 2-20 mg/L, vitamin B12: 2-20 mg/L.
6. The process for the production of microbial peptones from vegetable oils as claimed in claim 1, characterized in that: in the first step, the fermentation temperature of fermentation liquor in the fermentation period is 25-35 ℃; the ventilation amount during the fermentation is 0.5-2.5 vvm; during fermentation, the dissolved oxygen in the fermentation liquor is 5 to 50 percent through the coordination of ventilation and fermentation liquor stirring; adding ammonia water into the fermentation liquor during fermentation to maintain the pH value of the fermentation liquor at 6.2-8; and (3) feeding sterile vegetable oil into the fermentation liquor during the fermentation period to maintain the volume ratio of the total fat content in the fermentation liquor at 0.5-2%.
7. The process for the production of microbial peptones from vegetable oils as claimed in claim 6, characterized in that: in the first step, the fermentation temperature of fermentation liquor during fermentation is 29-31 ℃; the ventilation volume during the fermentation is one of three ranges of 1-1.5 vvm, 1.5-2 vvm and 1.8-2.5 vvm; adding 10-20% ammonia water into the fermentation liquor during fermentation to maintain the pH value of the fermentation liquor at 7.2-7.8; and (3) feeding sterile vegetable oil into the fermentation liquor during the fermentation period to maintain the volume ratio of the total fat content in the fermentation liquor at one of the two ranges of 0.5-1.5% and 1-2%.
8. The process for the production of microbial peptones from vegetable oils as claimed in claim 1 or 2 or 3 or 6, characterized in that: in the second step, when the fermentation liquor containing the single-cell protein is subjected to precipitation separation and centrifugation treatment in sequence, the centrifugation rotating speed is 5000 rpm-14000 rpm, and the centrifugation time is 3 minutes-20 minutes; when the high-pressure homogenizer carries out cell disruption treatment, the disruption pressure of the high-pressure homogenizer is 135MPa to 150 MPa; during spray drying treatment, the temperature of the spray airflow in the spray dryer is 175-185 ℃, and the temperature of the spray outlet in the spray dryer is 75-80 ℃.
9. The process for the production of microbial peptones from vegetable oils as claimed in claim 1 or 2 or 3 or 6, characterized in that: in the second step, the enzymolysis process is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
10. The process for the production of microbial peptones from vegetable oils as claimed in claim 8, characterized in that: in the second step, the enzymolysis process is as follows: firstly, adding a dissolving promoter into a fermentation liquor containing single-cell protein to carry out a first reaction, wherein the dissolving promoter is sodium chloride (calculated by 3% yeast solid solution) or ethanol, the reaction temperature of the first reaction is 50-60 ℃, the pH value of a reaction liquid in the first reaction process is maintained at 4.5-5.5, and the reaction time of the first reaction is 5-8 h; and after the first reaction is finished, adding papain into the first reaction solution to carry out a second reaction, wherein the adding mass of the papain is 0.5-0.8 per mill of the mass of the first reaction solution, the reaction temperature of the second reaction is 50-60 ℃, acetic acid is used for adjusting in the process of the second reaction to maintain the pH value of the second reaction solution at 5.5-6.5, and the reaction time of the second reaction is 15-20 hours.
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| CN110669797A (en) * | 2018-07-03 | 2020-01-10 | 上海凯赛生物技术股份有限公司 | Method for producing long-chain dicarboxylic acid by fermentation |
| CN110452825A (en) * | 2019-08-06 | 2019-11-15 | 苏州吉态来胺生物科技有限公司 | The fermentation process of single cell protein is produced using palm waste oil as raw material |
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Application publication date: 20201229 |