CN113201563B - A kind of nutrient salt and its application for improving the yield of Sclerotinia polysaccharide - Google Patents

Abstract

The invention provides a nutritive salt for improving the yield of sclerotium rolfsii polysaccharide and application thereof, belonging to the field of microbial fermentation. The nutrient salt for improving the yield of sclerotium rolfsii polysaccharide comprises the following components: disodium 6-phosphofructose, pyruvic acid, acetic acid, acetaldehyde and glutamic acid. The invention also provides application of the nutrient salt in producing sclerotium rolfsii polysaccharide. The nutritive salt is added at different time periods of fermentation and the pH values at different time periods are adjusted and controlled in a matching way, so that the synthesis of the sclerotium rolfsii polysaccharide is facilitated, and the aims of improving the yield of the sclerotium rolfsii polysaccharide, improving the utilization rate of a substrate and reducing the environmental pollution are fulfilled. The invention opens up a new way for improving the yield of the sclerotium rolfsii polysaccharide by adding the intermediate metabolite, can achieve more ideal polysaccharide yield compared with the prior art, overcomes the defects of complicated fermentation operation and high carbon source demand, and is beneficial to promoting the application and development of the sclerotium rolfsii polysaccharide in industry.

Description

A kind of nutrient salt and its application for improving the yield of Sclerotinia polysaccharide

technical field

The invention relates to the field of microbial fermentation, in particular to a nutrient salt for improving the yield of Sclerotinia polysaccharide and its application.

Background technique

Microbial exopolysaccharides are widely used in chemical industry, medicine, food, cosmetics, ecological protection and other fields due to their unique physical and chemical properties, rheological characteristics and biological safety. Sclerotin, also known as scleroglucan, is a microbial exopolysaccharide synthesized and secreted by some filamentous fungi of the genus Sclerotinia, among which the production of Sclerotin by fermentation of Sclerotinia is the most typical. Sclerotin has remarkable rheology and stability in environments with large pH, salinity and temperature changes. The field has broad application prospects.

However, the yield and yield of Sclerotin produced by fungal fermentation is not high, which leads to its high price, which seriously affects its application and development in industry. Therefore, how to increase the production of sclerotin is a current research hotspot.

At present, the methods for improving Sclerotin in the prior art mainly include the following three aspects: screening and mutagenesis of high-yield strains, selection of fermentation medium and optimization of fermentation conditions. For example, the Chinese non-patent literature "Production of Sclerotium polysaccharides by fermentation" discloses a high-yield strain Sclerotium rolfsii No.1, in a 16-liter self-controlled tank fermentation test, the polysaccharide yield reached 14.14g/L; Chinese patent literature CN108441429A discloses Sclerotium rolfsii WSH-G01, a Sclerotium rolfsii, was fermented for 56 hours under the condition of a glucose carbon source concentration of 75g/L to achieve a yield of 20g/L of scleroglucan. The polysaccharide yield of the above two schemes still needs to be improved. As another example, the Chinese non-patent literature "The Feed Control Strategy of Fermentation of Sclerotinia Sclerotin to Produce Scleroglucan" determined to control the stirring speed at 400r/min, start fermentation at 40h, and feed the total glucose concentration of 400g/L at a constant flow rate , the flow rate is 6g/(L h), and the feed feeding strategy is 32h. Although this method has improved the yield of polysaccharides, it needs to feed continuously for 32h during the fermentation process. The operation is complicated and the demand for carbon sources is high. .

To sum up, it is an urgent technical problem to be solved in this field to provide a solution that can increase the yield of Sclerotin, is easy to operate, and saves the amount of carbon source used.

Contents of the invention

Therefore, the technical problem to be solved by the present invention is to overcome the defects of low Sclerotin production, complex fermentation operation and high carbon source demand in the prior art, so as to provide a nutrient salt for increasing Sclerotin production and its application.

In the first aspect, the present invention provides a nutrient salt for improving the production of Sclerotin, which includes disodium fructose 6-phosphate, pyruvic acid, acetic acid, acetaldehyde and glutamic acid.

Further, the nutrient salts for increasing the yield of Sclerotin include, in parts by weight: 0.01-0.2 parts of disodium fructose 6-phosphate, 0.01-0.1 parts of pyruvate, 0.01-0.1 parts of acetic acid, acetaldehyde 0.01 to 0.1 parts and 0.01 to 0.1 parts of glutamic acid.

Further, the nutrient salts for increasing the yield of Sclerotin, in parts by weight, include: 0.06-0.15 parts of disodium fructose 6-phosphate, 0.01-0.1 parts of pyruvate, 0.05-0.1 parts of acetic acid, acetaldehyde 0.02 to 0.1 parts and 0.01 to 0.1 parts of glutamic acid.

Further, the nutrient salt for improving the yield of Sclerotin includes component A and component B, wherein, in parts by weight,

The component A includes: 0.01-0.1 parts of disodium fructose 6-phosphate, 0.01-0.1 parts of pyruvic acid and 0.01-0.1 parts of acetic acid;

The component B comprises: 0.05-0.1 part of disodium fructose 6-phosphate, 0.01-0.1 part of acetaldehyde and 0.01-0.1 part of glutamic acid.

Further, the component A includes: 0.01-0.05 parts of disodium fructose 6-phosphate, 0.01-0.1 parts of pyruvic acid and 0.05-0.1 parts of acetic acid;

The component B includes: 0.05-0.1 part of disodium fructose 6-phosphate, 0.02-0.1 part of acetaldehyde and 0.01-0.1 part of glutamic acid.

Further, the component A includes: 0.01 part of disodium fructose 6-phosphate, 0.01 part of pyruvic acid, and 0.05 part of acetic acid;

The component B includes: 0.05 part of disodium fructose 6-phosphate, 0.02 part of acetaldehyde and 0.01 part of glutamic acid.

Further, the component A includes: 0.05 part of disodium fructose 6-phosphate, 0.1 part of pyruvic acid and 0.1 part of acetic acid;

The component B includes: 0.1 part of disodium fructose 6-phosphate, 0.1 part of acetaldehyde and 0.1 part of glutamic acid.

In the second aspect, the present invention provides the application of the nutrient salt for increasing the yield of Sclerotin in the production of Sclerotin.

Further, the method for producing Sclerotin comprises: inoculating Sclerotinia neat into a fermentation medium for fermentation, and adding the nutrient salt to the fermentation medium during the fermentation process.

Further, the nutrient salts include: disodium fructose 6-phosphate 0.01-0.2g/L, pyruvate 0.01-0.1g/L, acetic acid 0.01-0.1g/L, acetaldehyde 0.01-0.1g/L and glutamine Acid 0.01～0.1g/L,

Further, the nutrient salt includes component A and component B, based on the volume of the fermentation medium,

The component A includes: disodium fructose 6-phosphate 0.01-0.1 g/L, pyruvic acid 0.01-0.1 g/L and acetic acid 0.01-0.1 g/L;

The component B includes: disodium fructose 6-phosphate 0.05-0.1g/L, acetaldehyde 0.01-0.1g/L and glutamic acid 0.01-0.1g/L,

Further, the component A includes: disodium fructose 6-phosphate 0.01g/L, pyruvate 0.01g/L, acetic acid 0.05g/L;

The component B includes: disodium fructose 6-phosphate 0.05g/L, acetaldehyde 0.02g/L, glutamic acid 0.01g/L,

Further, the component A includes: disodium fructose 6-phosphate 0.05g/L, pyruvate 0.1g/L and acetic acid 0.1g/L;

The component B includes: 0.1 g/L of disodium fructose 6-phosphate, 0.1 g/L of acetaldehyde and 0.1 g/L of glutamic acid.

Further, adding the component A to the fermentation medium during 0-12 hours of fermentation;

Adding the component B to the fermentation medium when fermenting for 12 to 24 hours,

Further, adding the component A to the fermentation medium when fermenting for 12 hours;

The component B was added to the fermentation medium at 24 hours of fermentation.

Further, the pH value of the fermentation medium is controlled to be 4±0.2 from the beginning of fermentation to 48h of fermentation;

The pH of the fermentation medium is controlled to be 3±0.2 from 48 hours of fermentation to the end of fermentation.

Further, the fermentation temperature is 28° C., the rotation speed of the shaker is 220 rpm, and the fermentation time is 72 hours.

Further, based on the volume of the fermentation medium, the fermentation medium includes: 75-100 g/L of sucrose or glucose, 0.5-1.5 g/L of yeast extract, 2.25-3 g/L of sodium nitrate, 1 g/L of dipotassium hydrogen phosphate ～2g/L, magnesium sulfate heptahydrate 0.4～0.5g/L, potassium chloride 0.4～0.5g/L, ferrous sulfate 0～0.05g/L and citric acid 0.7～1.5g/L,

Further, the fermentation medium comprises:

Sucrose 100g/L, yeast extract 1g/L, sodium nitrate 2.25g/L, dipotassium hydrogen phosphate 2g/L, magnesium sulfate heptahydrate 0.5g/L, potassium chloride 0.5g/L, ferrous sulfate 0.05g/L L and citric acid 0.7g/L; or

Glucose 75g/L, yeast extract 1g/L, sodium nitrate 2.25g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate heptahydrate 0.5g/L, potassium chloride 0.5g/L and citric acid 1.5g/L ;or

Glucose 95g/L, yeast extract 1g/L, sodium nitrate 3g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate heptahydrate 0.5g/L, potassium chloride 0.5g/L and citric acid 1.5g/L.

The technical solution of the present invention has the following advantages:

1. The nutrient salts that improve the output of Sclerotin provided by the present invention include disodium fructose 6-phosphate, pyruvic acid, acetic acid, acetaldehyde and glutamic acid, by introducing nutrient salts in the fermentation medium, Sclerotin is made Production has been greatly increased.

2. Based on the staged characteristics of the whole fermentation process, the nutrient salt is further divided into component A and component B, and the nutrient salt supplement is carried out through different fermentation stages, which is more in line with the synthesis law of Sclerotinia polysaccharide and is conducive to further improvement Sclerotin production.

3. The present invention also provides the application of nutrient salts for improving the yield of Sclerotin in the production of Sclerotin. Adding nutrient salts in different periods of fermentation and cooperating with regulating the pH value in different periods is beneficial to promote the production of Sclerotin. Synthesis, the output can reach 34.1g/L, and then achieve the purpose of increasing the yield of Sclerotin, increasing the utilization rate of substrates and reducing environmental pollution.

4. The present invention provides a brand-new scheme for increasing the yield of Sclerotinia polysaccharide, and opens up a new way to increase the yield of Sclerotinia polysaccharide by adding intermediate metabolites, which can achieve a relatively ideal polysaccharide yield compared with the prior art , and overcome the defects of complex fermentation operation and high carbon source demand, which is conducive to promoting the application and development of Sclerotin in industry.

Detailed ways

The following examples are provided in order to further understand the present invention better, are not limited to the best implementation mode, and do not limit the content and protection scope of the present invention, anyone under the inspiration of the present invention or use the present invention Any product identical or similar to the present invention obtained by combining features of other prior art falls within the protection scope of the present invention.

If no specific experimental steps or conditions are indicated in the examples, it can be carried out according to the operation or conditions of the conventional experimental steps described in the literature in this field. The raw materials or instruments used are commercially available conventional products, including but not limited to the raw materials or instruments used in the examples of this application.

The raw material source used in the embodiment:

The strain of Sclerotium rolfsii used in the examples is Sclerotium rolfsii ATCC15205, which was purchased from China Industrial Microorganism Culture Collection and Management Center.

The PDA liquid medium used in the examples was prepared by the following method: 200g of potatoes, peeled, cut into pieces, added distilled water and boiled for 30min, filtered to obtain the filtrate, added 20g of glucose, and replenished with water to 1000mL.

Other raw materials used in the embodiment and their specifications and sources are shown in Table 1.

Table 1 Specifications and sources of raw materials

Example 1

Improve the nutrient salt of sclerotin and its application in the production of sclerotin:

Inoculate the mature Sclerotinia sclerotinum liquid into PDA liquid medium (100mL/250mL), the inoculation amount is 5%, 28°C, 220rpm, cultivate for 72h, and obtain a seed liquid containing a large amount of hyphae;

The seed liquid obtained is inoculated in the fermentation medium according to the inoculation amount of 5%, 28 ℃, 220rpm, cultivate 72h, wherein, add the component A of nutrient salt in the fermentation medium when fermentation starts, when fermenting 12h, add to Component B of nutrient salts is added to the fermentation medium, and the pH value of the fermentation medium is controlled to be 4±0.2 from the beginning of fermentation to 48 hours of fermentation, and the pH value of the fermentation medium is controlled to be 3±0.2 from 48 hours of fermentation to the end of fermentation.

In this example, the composition of the fermentation medium is: sucrose 100g/L, yeast extract 1g/L, sodium nitrate (NaNO ₃ ) 2.25g/L, dipotassium hydrogen phosphate (K ₂ HPO ₄ ) 2g/L, Magnesium sulfate heptahydrate (MgSO ₄ ·7H ₂ O) 0.5g/L, potassium chloride (KCl) 0.5g/L, ferrous sulfate (FeSO ₄ ) 0.05g/L, citric acid 0.7g/L.

In this embodiment, the nutrient salt is composed of component A and component B,

Component A: 0.01 g of disodium fructose 6-phosphate, 0.01 g of pyruvate, and 0.05 g of acetic acid were added to each 1 L of fermentation medium;

Component B: 0.05 g of disodium fructose 6-phosphate, 0.02 g of acetaldehyde, and 0.01 g of glutamic acid were added to 1 L of fermentation medium.

Comparative example 1-1

The seed solution obtained in Example 1 was inoculated in the fermentation medium of Example 1 according to the inoculation amount of 5%, and cultivated for 72 hours at 28°C and 220rpm. No nutrient salt was added to the fermentation medium during the cultivation process, nor was it adjusted. pH manipulation.

Comparative example 1-2

The seed liquid obtained in Example 1 was inoculated in the fermentation medium of Example 1 according to an inoculum size of 5%, cultivated for 72 hours at 28° C., 220 rpm, and no nutrient salt was added to the fermentation medium during the cultivation process. The pH value of the fermentation medium was controlled to be 4±0.2 before 48 hours of fermentation, and the pH value of the fermentation medium was controlled to be 3±0.2 during 48 hours of fermentation until the end of fermentation.

Example 2

Improve the nutrient salt of sclerotin and its application in the production of sclerotin:

The method for obtaining seed liquid is the same as in Example 1;

The obtained seed liquid is inoculated in the fermentation medium according to the inoculum amount of 5%, cultivated for 72 hours at 28° C. at 220 rpm, wherein, when fermenting for 12 hours, component A of nutrient salt is added to the fermentation medium, and when fermenting for 24 hours, it is added to Component B of nutrient salts is added to the fermentation medium, and the pH value of the fermentation medium is controlled to be 4±0.2 from the beginning of fermentation to 48 hours of fermentation, and the pH value of the fermentation medium is controlled to be 3±0.2 from 48 hours of fermentation to the end of fermentation.

In this example, the composition of the fermentation medium is: glucose 75g/L, yeast extract 1g/L, sodium nitrate (NaNO ₃ ) 2.25g/L, dipotassium hydrogen phosphate (K ₂ HPO ₄ ) 1g/L, Magnesium sulfate heptahydrate (MgSO ₄ ·7H ₂ O) 0.5g/L, potassium chloride (KCl) 0.5g/L, citric acid 1.5g/L.

In this embodiment, the nutrient salt is composed of component A and component B,

Component A: Add 0.025 g of disodium fructose 6-phosphate, 0.05 g of pyruvate, and 0.05 g of acetic acid into each 1 L of fermentation medium;

Component B: 0.05 g of disodium fructose 6-phosphate, 0.025 g of acetaldehyde and 0.05 g of glutamic acid were added to each 1 L of fermentation medium.

Comparative example 2-1

The seed solution obtained in Example 1 was inoculated into the fermentation medium of Example 2 according to the inoculation amount of 5%, and cultivated for 72 hours at 28°C and 220rpm. No nutrient salt was added to the fermentation medium during the cultivation process, nor was it adjusted. pH manipulation.

Comparative example 2-2

The seed liquid obtained in Example 1 was inoculated in the fermentation medium of Example 2 according to an inoculum size of 5%, and cultivated for 72 hours at 28° C. at 220 rpm. No nutrient salt was added to the fermentation medium during the cultivation process. The pH value of the fermentation medium was controlled to be 4±0.2 before 48 hours of fermentation, and the pH value of the fermentation medium was controlled to be 3±0.2 during 48 hours of fermentation until the end of fermentation.

Example 3

Improve the nutrient salt of sclerotin and its application in the production of sclerotin:

The method for obtaining seed liquid is the same as in Example 1;

The obtained seed liquid is inoculated in the fermentation medium according to the inoculum amount of 5%, cultivated for 72 hours at 28° C. at 220 rpm, wherein, when fermenting for 12 hours, component A of nutrient salt is added to the fermentation medium, and when fermenting for 24 hours, it is added to Component B of nutrient salts is added to the fermentation medium, and the pH value of the fermentation medium is controlled to be 4±0.2 from the beginning of fermentation to 48 hours of fermentation, and the pH value of the fermentation medium is controlled to be 3±0.2 from 48 hours of fermentation to the end of fermentation.

In this example, the composition of the fermentation medium is: glucose 95g/L, yeast extract 1g/L, sodium nitrate (NaNO ₃ ) 3g/L, dipotassium hydrogen phosphate (K ₂ HPO ₄ ) 1g/L, seven Magnesium sulfate water (MgSO ₄ ·7H ₂ O) 0.5g/L, potassium chloride (KCl) 0.5g/L, citric acid 1.5g/L.

In this embodiment, the nutrient salt is composed of component A and component B,

Component A: add 0.05 g of disodium fructose 6-phosphate, 0.1 g of pyruvic acid, and 0.1 g of acetic acid into each 1 L of fermentation medium;

Component B: 0.1 g of disodium fructose 6-phosphate, 0.1 g of acetaldehyde, and 0.1 g of glutamic acid were added to each 1 L of fermentation medium.

Comparative example 3-1

The seed solution obtained in Example 1 was inoculated into the fermentation medium of Example 3 according to the inoculation amount of 5%, and cultivated for 72 hours at 28°C and 220rpm. No nutrient salt was added to the fermentation medium during the cultivation process, and no adjustment was made. pH manipulation.

Comparative example 3-2

The seed liquid obtained in Example 1 was inoculated in the fermentation medium of Example 3 according to an inoculum size of 5%, cultivated for 72 hours at 28° C., 220 rpm, and no nutrient salt was added to the fermentation medium during the cultivation process. The pH value of the fermentation medium was controlled to be 4±0.2 before 48 hours of fermentation, and the pH value of the fermentation medium was controlled to be 3±0.2 during 48 hours of fermentation until the end of fermentation.

Example 4

Improve the nutrient salt of sclerotin and its application in the production of sclerotin:

The method for obtaining seed liquid is the same as in Example 1;

The obtained seed liquid is inoculated in the fermentation medium according to the inoculum amount of 5%, cultivated for 72 hours at 28° C. at 220 rpm, wherein, when fermenting for 12 hours, component A of nutrient salt is added to the fermentation medium, and when fermenting for 24 hours, it is added to Component B of nutrient salt is added to the fermentation medium, and the pH value of the fermentation medium is controlled to be 3±0.2 from the beginning of the fermentation to the end of the fermentation.

In this embodiment, the composition of the fermentation medium and the composition of the nutrient salt are the same as those in Embodiment 3.

Example 5

Improve the nutrient salt of sclerotin and its application in the production of sclerotin:

The method for obtaining seed liquid is the same as in Example 1;

The obtained seed liquid was inoculated in the fermentation medium according to the inoculum amount of 5%, cultivated for 72 hours at 28° C. and 220 rpm, wherein, when the fermentation was 12 hours, component A and component B of nutrient salts were added to the fermentation medium at the same time, The pH value of the fermentation medium was controlled to be 4±0.2 from the beginning of the fermentation to 48 hours of fermentation, and the pH value of the fermentation medium was controlled to be 3±0.2 during 48 hours of fermentation to the end of the fermentation.

In this embodiment, the composition of the fermentation medium and the composition of the nutrient salt are the same as those in Embodiment 3.

Experimental example

Sampling was carried out on the fermented liquid when fermented for 72 hours in Examples and Comparative Examples, and Sclerotin was extracted and its content was determined.

Extraction method of Sclerotin: After 72 hours of fermentation, filter the fermentation liquid to obtain the fermentation filtrate, add four times the volume of absolute ethanol, alcohol precipitation overnight at 4°C, and centrifuge at 5000rmp for 20 minutes to obtain the precipitate, redissolve the precipitate with water, and obtain the Test samples.

The content determination method of Sclerotin: phenol-sulfuric acid method, the determination system is 1.00mL of the sample to be tested, add 1.0mL 5% phenol solution, then quickly add 5.0mL concentrated sulfuric acid in an ice bath, let it stand at room temperature for 30min, measure the absorbance at 490nm , calculate the concentration of Sclerotin according to the standard curve, wherein,

Standard curve: y=9.0153x- ^0.0129R2 =0.9963

y-absorbance x-sclerotin concentration

Calculate the Sclerotin weight according to the measured Sclerotin concentration and the volume of the sample to be tested, and calculate the Sclerotin output according to the following formula:

Sclerotin yield=(Sclerotin weight/fermentation broth volume)×100%.

The yields of Sclerotin obtained in each example and comparative example are shown in Table 2.

Table 2 Sclerotin production

Sclerotin production (g/L) Example 1 32.2 Comparative example 1-1 21.7 Comparative example 1-2 29.4 Example 2 25.4 Comparative example 2-1 18.7 Comparative example 2-2 22.3 Example 3 34.1 Comparative example 3-1 21.5 Comparative example 3-2 29.6 Example 4 23.4 Example 5 31.2

As can be seen from Table 2, compared with the scheme (comparative example 1-1) that does not add nutrient salts and does not adjust the pH value in the fermentation process, the output of Example 1 increases by 48.4%. For the scheme of pH value (comparative example 1-2), the yield increased by 9.5%; compared with the scheme (comparative example 2-1) in which no nutrient salt was added and the pH value was not adjusted in the fermentation process in Example 2, the yield increased by 35.8%, Compared with the scheme (comparative example 2-2) that does not add nutrient salts in the fermentation process and adjusts the pH value accordingly, the yield increases by 13.9%; compared with the scheme (comparative example 2-2) that does not add nutrient salts and does not adjust the pH value in the fermentation process ( Comparative example 3-1), the output increased by 58.6%, compared with the scheme (comparative example 3-2) that did not add nutrient salt in the fermentation process, and correspondingly adjusted the pH value, the output increased by 15.2%; The scheme (Example 4) of adjusting the pH value increased the yield by 45.7%; compared with the scheme (Example 5) of adding component A and component B simultaneously in Example 3, the yield increased by 9.3%.

This proves that adopting the nutrient salt provided by the present invention can effectively increase the yield of Sclerotin, and the yield can be further increased in conjunction with the stage-by-stage regulation of the pH value. In addition, compared with the one-time addition of nutrient salts, the staged addition of components A and B can also improve the yield to a certain extent.

Apparently, the above-mentioned embodiments are only examples for clear description, rather than limiting the implementation. For those of ordinary skill in the art, other changes or changes in different forms can be made on the basis of the above description. It is not necessary and impossible to exhaustively list all the implementation manners here. And the obvious changes or changes derived therefrom are still within the scope of protection of the present invention.

Claims (6)

1. The application of a nutritive salt for improving the yield of sclerotium rolfsii polysaccharide in the production of sclerotium rolfsii polysaccharide is characterized in that the method for producing sclerotium rolfsii polysaccharide comprises the following steps: inoculating sclerotium rolfsii into a fermentation culture medium for fermentation, adding the nutrient salt into the fermentation culture medium in the fermentation process,

the nutrient salt comprises a component A and a component B, based on the volume of the fermentation medium,

the component A comprises: 0.01-0.1 g/L of 6-disodium fructose phosphate, 0.01-0.1 g/L of pyruvic acid and 0.01-0.1 g/L of acetic acid;

the component B comprises: 0.05-0.1 g/L of 6-disodium fructose phosphate, 0.01-0.1 g/L of acetaldehyde and 0.01-0.1 g/L of glutamic acid,

adding the component A into the fermentation medium when fermenting for 0-12 h;

adding the component B into the fermentation medium when fermenting for 12-24 h,

controlling the pH value of the fermentation medium to be 4 +/-0.2 from the beginning of fermentation to 48 hours before fermentation;

controlling the pH value of the fermentation medium to be 3 +/-0.2 from 48h to the end of fermentation.

2. The use according to claim 1,

the component A comprises: 0.01g/L of 6-disodium fructose phosphate, 0.01g/L of pyruvic acid and 0.05g/L of acetic acid;

the component B comprises: 0.05g/L of 6-disodium fructose phosphate, 0.02g/L of acetaldehyde, 0.01g/L of glutamic acid, or

The component A comprises: 0.05g/L of 6-disodium fructose phosphate, 0.1g/L of pyruvic acid and 0.1g/L of acetic acid;

the component B comprises: 0.1g/L of 6-disodium fructose phosphate, 0.1g/L of acetaldehyde and 0.1g/L of glutamic acid.

3. The use according to claim 1,

adding the component A into the fermentation medium during fermentation for 12 h;

the component B was added to the fermentation medium at the time of fermentation for 24 h.

4. The use according to claim 1, wherein the fermentation temperature is 28 ℃, the shaker speed is 220rpm, and the fermentation time is 72h.

5. Use according to claim 1, wherein the fermentation medium comprises, based on the volume of the fermentation medium: 75-100 g/L of sucrose or glucose, 0.5-1.5 g/L of yeast extract, 2.25-3 g/L of sodium nitrate, 1-2 g/L of dipotassium phosphate, 0.4-0.5 g/L of magnesium sulfate heptahydrate, 0.4-0.5 g/L of potassium chloride, 0-0.05 g/L of ferrous sulfate and 0.7-1.5 g/L of citric acid.

6. Use according to claim 5, wherein the fermentation medium comprises, based on the volume of the fermentation medium:

100g/L of sucrose, 1g/L of yeast extract, 2.25g/L of sodium nitrate, 2g/L of dipotassium phosphate, 0.5g/L of magnesium sulfate heptahydrate, 0.5g/L of potassium chloride, 0.05g/L of ferrous sulfate and 0.7g/L of citric acid; or

75g/L glucose, 1g/L yeast extract, 2.25g/L sodium nitrate, 1g/L dipotassium phosphate, 0.5g/L magnesium sulfate heptahydrate, 0.5g/L potassium chloride and 1.5g/L citric acid; or

95g/L glucose, 1g/L yeast extract, 3g/L sodium nitrate, 1g/L dipotassium phosphate, 0.5g/L magnesium sulfate heptahydrate, 0.5g/L potassium chloride and 1.5g/L citric acid.