CN110564804A - Clear liquid fermentation medium for producing riboflavin and fermentation method - Google Patents
Clear liquid fermentation medium for producing riboflavin and fermentation method Download PDFInfo
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- CN110564804A CN110564804A CN201910866956.2A CN201910866956A CN110564804A CN 110564804 A CN110564804 A CN 110564804A CN 201910866956 A CN201910866956 A CN 201910866956A CN 110564804 A CN110564804 A CN 110564804A
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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Abstract
The invention relates to a clear liquid fermentation medium and a fermentation method for producing riboflavin, belonging to the technical field of biological pharmacy, wherein the clear liquid fermentation medium comprises the following components in percentage by weight: 20-30g/L of corn steep liquor dry powder, 15-25g/L of yeast extract, 0.5-1.0g/L of magnesium sulfate, 2-5g/L of betaine, 2-5g/L of dipotassium phosphate and 3-6g/L of diammonium phosphate. The clear liquid fermentation culture medium optimizes the traditional culture medium, reasonably prepares components, reduces the cost of raw materials, and obviously improves the yield of riboflavin by utilizing the clear liquid fermentation culture medium for production.
Description
Technical Field
The invention belongs to the technical field of biological pharmacy, and particularly relates to a clear liquid fermentation medium for producing riboflavin and a fermentation method.
Background
Riboflavin is also known as VB2, also known as vitamin G or lactoflavin, and belongs to water-soluble. It is indispensable to biological life activities, is an essential component constituting flavoenzymes, and directly participates in carbohydrate, protein and fat metabolism. If the food lacks riboflavin, a human body may develop a series of health problems due to the long-term lack of riboflavin. The method for producing riboflavin by fermentation has the advantages of mild reaction conditions, low cost, greenness, cleanness, environmental protection and the like. Therefore, the production of riboflavin by fermentation under the condition of low carbon and environmental protection has important significance.
The existing riboflavin production process has the disadvantages of complex fermentation material components, low conversion rate, long fermentation period and low yield, and simultaneously has serious sewage pollution after extraction, so that the production cost is high and the environmental cost is extremely high due to the influence of various aspects, which is mainly caused by the fermentation production process. Moreover, in the prior art, riboflavin is industrially produced by adopting a traditional culture medium for fermentation production, but the traditional culture medium generally contains cottonseed cake powder and beet molasses, the two components can provide nutrient substances required by growth and development for fermenting organisms, but long-term practice production and research show that the two components have some defects in the fermentation process, the solubility of the cottonseed cake powder is low, and the use of the cottonseed cake powder as a fermentation nitrogen source not only makes the fermentation liquor viscous but also reduces the concentration of dissolved oxygen in the fermentation liquor; the beet molasses contains more colloidal substances in the components, so that the fermentation liquor has early foaming time and larger foam in the fermentation process. These seriously affect the cycle time and yield of riboflavin production.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims at providing a clear liquid fermentation culture medium for producing riboflavin and aims at providing a method for producing riboflavin by fermentation by using the clear liquid fermentation culture medium. The clear liquid fermentation culture medium optimizes the traditional culture medium, reasonably prepares components, reduces the cost of raw materials, and obviously improves the yield of riboflavin.
In order to achieve the purpose, the invention adopts the specific scheme that:
A clear liquid fermentation medium for producing riboflavin comprises the following components in concentration: 20-30g/L of corn steep liquor dry powder, 15-25g/L of yeast extract, 0.5-1.0g/L of magnesium sulfate, 2-5g/L of betaine, 2-5g/L of dipotassium phosphate and 3-6g/L of diammonium phosphate.
As a further optimization of the above scheme, the components and their concentrations contained in the clear liquid fermentation medium are respectively: 25g/L of corn steep liquor dry powder, 20g/L of yeast extract, 0.8g/L of magnesium sulfate, 3g/L of betaine, 3g/L of dipotassium phosphate and 6g/L of diammonium phosphate.
The invention also provides a fermentation method for producing riboflavin, which inoculates riboflavin-producing bacteria bacillus subtilis into a clear liquid fermentation culture medium for fermentation culture under the following culture conditions: at 42 deg.C, pH7.2, 0.8m3Culturing at constant temperature of 0.5-1.0% and 0.05Mpa for 60h with dissolved oxygen controlled at 30-40% and residual sugar controlled at 60.5-1.0%.
has the advantages that:
1. The invention develops a new process for clear liquid fermentation culture on the premise of the existing riboflavin production strains, on one hand, the cost of raw materials is reduced, and the utilization rate of equipment is improved; on the other hand, the extraction yield is improved after the fermentation liquor quality is improved, and the product quality is ensured. The invention provides a new process for clear fermentation culture of riboflavin, which has the advantages of mild reaction conditions, low cost, greenness, cleanness, environmental protection and the like.
2. The clear liquid fermentation culture medium is optimized by the traditional fermentation culture medium, unnecessary components such as cottonseed cake powder, beet molasses and the like are removed, and the clear liquid culture medium containing components such as corn steep liquor dry powder, yeast extract, magnesium sulfate, betaine, dipotassium hydrogen phosphate, diammonium hydrogen phosphate and the like is adopted. The invention uses yeast extract to replace cottonseed cake powder, the yeast extract is a full-solubility nitrogen source, and the amino acids and the nucleotide substances in the components are more abundant than the cottonseed cake powder; the betaine is one of the main components of the beet molasses, and the invention uses the betaine to replace the beet molasses, not only retains the main components, but also removes colloidal substances which have adverse effect on riboflavin fermentation, and further improves the quality of fermentation liquor.
3. Compared with the traditional fermentation method, the method has the beneficial effects that: the fermentation method disclosed by the invention is adopted for fermentation for 60 hours, the yield of the riboflavin reaches 30-40g/L, the fermentation period is short, the yield is higher, the reaction condition is mild, the cost is low, and the fermentation method has the advantages of greenness, cleanness, environmental protection and the like.
Detailed Description
The technical scheme of the invention is further described by combining the specific examples of small test and large production.
Example 1
Carrying out fermentation culture by utilizing riboflavin-producing bacteria bacillus subtilis by adopting a clear liquid fermentation culture medium, wherein the clear liquid fermentation culture medium comprises the following components in percentage by weight: the addition amount of the corn steep liquor dry powder is 20g/L, the addition amount of the yeast extract is 15g/L, the addition amount of magnesium sulfate is 0.5g/L, the addition amount of betaine is 5g/L, the addition amount of dipotassium hydrogen phosphate is 2g/L and the addition amount of diammonium hydrogen phosphate is 6 g/L. The culture conditions are as follows: culturing at 42 deg.C, pH7.2, 0.8m3/h, 0.05Mpa, dissolved oxygen 30-40%, and residual sugar 0.5-1.0, fermenting in 50L fermenter for 60h, and riboflavin yield 35 g/L.
Example 2
Carrying out fermentation culture by utilizing riboflavin-producing bacteria bacillus subtilis by adopting a clear liquid fermentation culture medium, wherein the clear liquid fermentation culture medium comprises the following components in percentage by weight: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the yeast extract is 20g/L, the addition amount of magnesium sulfate is 0.8g/L, the addition amount of betaine is 3g/L, the addition amount of dipotassium hydrogen phosphate is 3g/L, and the addition amount of diammonium hydrogen phosphate is 6 g/L. The culture conditions were: culturing at 42 deg.C, pH7.2, 0.8m3/h, 0.05Mpa, dissolved oxygen 30-40%, and residual sugar 0.5-1.0, fermenting in a small-scale 50L fermentation tank for 60h, and increasing riboflavin yield to 38 g/L.
Example 3
Carrying out fermentation culture by utilizing riboflavin-producing bacteria bacillus subtilis by adopting a clear liquid fermentation culture medium, wherein the clear liquid fermentation culture medium comprises the following components in percentage by weight: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the yeast extract is 20g/L, the addition amount of magnesium sulfate is 0.8g/L, the addition amount of betaine is 3g/L, the addition amount of dipotassium hydrogen phosphate is 3g/L, and the addition amount of diammonium hydrogen phosphate is 6 g/L. The fermentation period of a large-scale production 130 cubic fermentor is 60 hours, and the yield of riboflavin is 32 g/L.
Example 4
Carrying out fermentation culture by utilizing riboflavin-producing bacteria bacillus subtilis by adopting a clear liquid fermentation culture medium, wherein the clear liquid fermentation culture medium comprises the following components in percentage by weight: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the yeast extract is 20g/L, the addition amount of magnesium sulfate is 0.8g/L, the addition amount of betaine is 3g/L, the addition amount of dipotassium hydrogen phosphate is 3g/L, and the addition amount of diammonium hydrogen phosphate is 6 g/L. The fermentation period of the 136 cubic fermentor in the production plant is 60 hours, and the yield of the riboflavin is 35 g/L.
Comparative example 1
The method comprises the following steps of (1) carrying out fermentation culture on riboflavin-producing bacteria bacillus subtilis by adopting a traditional culture medium, wherein the traditional culture medium comprises the following components in concentration: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the cottonseed cake powder is 20g/L, the addition amount of the magnesium sulfate is 0.8g/L, the addition amount of the beet molasses is 20g/L, the addition amount of the dipotassium phosphate is 3g/L and the addition amount of the diammonium phosphate is 6 g/L. The culture conditions were: at 42 deg.C, pH7.2, 0.8m3H, 0.05Mpa, dissolved oxygen control of 30-40%, residual sugar control of 0.5-1.0 constant culture, small test 50L fermentation tank fermentation period of 60h, and riboflavin output of 25 g/L.
Comparative example 2
The method comprises the following steps of (1) carrying out fermentation culture on riboflavin-producing bacteria bacillus subtilis by adopting a traditional culture medium, wherein the traditional culture medium comprises the following components in concentration: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the cottonseed cake powder is 20g/L, the addition amount of the magnesium sulfate is 0.8g/L, the addition amount of the beet molasses is 20g/L, the addition amount of the dipotassium phosphate is 3g/L, and the addition amount of the diammonium phosphate is 6 g/L. The culture conditions were: at 42 deg.C, pH7.2, 0.8m3H, 0.05Mpa, dissolved oxygen control of 30-40 percent and residual sugar control of 0.5-1.0 constant culture, the fermentation period of a large-scale production 130 cubic fermentation tank is 60h, and the yield of riboflavin is 23 g/L.
Comparative example 3
The method comprises the following steps of (1) carrying out fermentation culture on riboflavin-producing bacteria bacillus subtilis by adopting a traditional culture medium, wherein the traditional culture medium comprises the following components in concentration: the addition amount of the corn steep liquor dry powder is 25g/L, the addition amount of the cottonseed cake powder is 20g/L, the addition amount of the magnesium sulfate is 0.8g/L, the addition amount of the beet molasses is 20g/L, the addition amount of the dipotassium phosphate is 3g/L, and the addition amount of the diammonium phosphate is 6 g/L. The fermentation period of the 136 cubic fermentor in the production plant is 60 hours, and the yield of the riboflavin is 20 g/L.
It should be noted that the above-mentioned embodiments illustrate rather than limit the scope of the invention, which is defined by the appended claims. It will be apparent to those skilled in the art that certain insubstantial modifications and adaptations of the present invention can be made without departing from the spirit and scope of the invention.
Claims (3)
1. A clear fermentation medium for the production of riboflavin, characterized in that: the clear liquid fermentation medium comprises the following components in percentage by weight: 20-30g/L of corn steep liquor dry powder, 15-25g/L of yeast extract, 0.5-1.0g/L of magnesium sulfate, 2-5g/L of betaine, 2-5g/L of dipotassium phosphate and 3-6g/L of diammonium phosphate.
2. A clear fermentation medium for the production of riboflavin according to claim 1, characterized in that: the clear liquid fermentation medium comprises the following components in percentage by weight: 25g/L of corn steep liquor dry powder, 20g/L of yeast extract, 0.8g/L of magnesium sulfate, 3g/L of betaine, 3g/L of dipotassium phosphate and 6g/L of diammonium phosphate.
3. A fermentation process for producing riboflavin using the clear fermentation medium according to claim 1 or 2, characterized in that: the method comprises the steps of inoculating the riboflavin-producing bacteria bacillus subtilis into a clear liquid fermentation culture medium for fermentation culture, wherein the culture conditions are as follows: at 42 deg.C, pH7.2, 0.8m3culturing at constant temperature of 0.5-1.0% and 0.05Mpa for 60h with dissolved oxygen controlled at 30-40% and residual sugar controlled at 60.5-1.0%.
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CN113249261A (en) * | 2021-05-26 | 2021-08-13 | 浙江新和成股份有限公司 | Bacillus subtilis and application thereof in production of riboflavin |
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