CN119242513B - Fermentation medium for improving surfactant content, application and fermentation method thereof - Google Patents
Fermentation medium for improving surfactant content, application and fermentation method thereof Download PDFInfo
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Abstract
本申请提供一种提高表面活性素含量的发酵培养基、其应用及发酵方法。发酵培养基用于枯草芽孢杆菌的发酵；发酵培养基包括以下重量百分比的物质：碳源4‑7％；氮源0.65‑2.25％；磷酸盐0.13‑0.6％；硫酸镁0.015‑0.05％；氯化钙0.0007‑0.02％；硫酸锰0.0005‑0.001％；硫酸铁0.0006‑0.06％；氨基酸0.5‑1％；大豆纤维0.5‑3％，余量为水；其中，发酵培养基的pH为6.5‑7；发酵培养基中枯草芽孢杆菌菌液的接种量以体积分数计为3‑5％；其中，碳源选自糖蜜、葡萄糖、红糖、麦芽糊精与淀粉中的至少一种；氮源包括玉米浆干粉、酵母膏和酵母粉中的至少一种与尿素的组合。
The present application provides a fermentation medium for increasing the content of surfactin, its application and fermentation method. The fermentation medium is used for fermentation of Bacillus subtilis; the fermentation medium includes the following substances in weight percentage: 4-7% carbon source; 0.65-2.25% nitrogen source; 0.13-0.6% phosphate; 0.015-0.05% magnesium sulfate; 0.0007-0.02% calcium chloride; 0.0005-0.001% manganese sulfate; 0.0006-0.06% iron sulfate; 0.5-1% amino acid; 0.5-3% soybean fiber, and the balance is water; wherein the pH of the fermentation medium is 6.5-7; the inoculation amount of Bacillus subtilis liquid in the fermentation medium is 3-5% by volume fraction; wherein the carbon source is selected from at least one of molasses, glucose, brown sugar, maltodextrin and starch; the nitrogen source includes a combination of at least one of corn slurry powder, yeast extract and yeast powder and urea.
Description
Technical Field
The application relates to the technical field of fermentation engineering, in particular to a fermentation medium for improving the content of surfactant, application thereof and a fermentation method.
Background
Biosurfactants are metabolites produced by microorganisms such as bacteria, yeasts, fungi from various substrates (sugars, oils, alkanes, etc.) and having both hydrophobicity and hydrophilicity, which can reduce surface and interfacial tension and produce micelles, enhancing the solubility of hydrocarbons in water or water in hydrocarbons. Biosurfactants have received extensive attention in recent years due to their low toxicity, surface activity and environmental friendliness.
Surfactants (Surfactin) are one of the most effective lipopeptid biosurfactants produced by various strains of bacillus subtilis (Bacillus subtilis). The main structure is a cyclic lipopeptide which is formed by connecting 12-16 carbon chain length fatty acids and 7 amino acids through internal ester bonds. The surfactant has antibacterial, antiviral, antitumor and fibrin clot formation inhibiting effects in addition to high surface activity, low toxicity, high biodegradability and biocompatibility. These excellent properties make it useful in the petroleum industry, environmental control has wide application prospect in the fields of cosmetics, agriculture, medicine and the like.
In the related art, the yield of the surfactant is low.
Disclosure of Invention
In view of the above, the present application aims to provide a fermentation medium for increasing the content of surfactant, its application and fermentation method.
Based on the purposes, the application provides a fermentation medium for improving the content of surface active substances, which is used for fermenting bacillus subtilis, and comprises, by weight, 4-7% of carbon source, 0.65-2.25% of nitrogen source, 0.13-0.6% of phosphate, 0.015-0.05% of magnesium sulfate, 0.0007-0.02% of calcium chloride, 0.0005-0.001% of manganese sulfate, 0.0006-0.06% of ferric sulfate, 0.5-1% of amino acid, 0.5-3% of soybean fibers and the balance of water, wherein the pH of the fermentation medium is 6.5-7, and the inoculation amount of bacillus subtilis bacterial liquid in the fermentation medium is 3-5% in terms of volume fraction;
The carbon source is at least one selected from molasses, glucose, brown sugar, maltodextrin and starch, and the nitrogen source comprises at least one of corn steep liquor dry powder, yeast extract and yeast powder in combination with urea.
In some embodiments, the soybean fiber is added in an amount of 1-3%.
In some embodiments, the nitrogen source is selected from the group consisting of yeast extract and urea, wherein the yeast extract is 0.45-0.55% by mass and the urea is 0.45-0.55% by mass.
In some embodiments, the carbon source is selected from brown sugar, wherein the mass percentage of the brown sugar is 4-7%.
In some of these embodiments, the phosphate is selected from the group consisting of potassium dihydrogen phosphate in combination with disodium hydrogen phosphate and the amino acid is selected from the group consisting of leucine.
The embodiment of the application also provides an application of the fermentation medium for improving the content of the surfactant in the fermentation product of bacillus subtilis.
The embodiment of the application also provides a fermentation method for improving the content of the surfactant, which comprises the following steps:
inoculating bacillus subtilis into a liquid seed culture medium, and carrying out seed culture for 16-18h to obtain fermentation strains;
Inoculating a fermentation strain into a fermentation culture medium for fermentation culture for 22-26 hours, wherein the inoculation amount of the fermentation strain is 3-5% in terms of volume fraction, and the fermentation culture medium is the fermentation culture medium in any embodiment;
And adding a feed medium, and carrying out feed culture for 46-50h.
In some embodiments, the feed medium comprises brown sugar, urea and leucine, and the feed medium is added in an amount such that the ratio of the added volume of the feed medium to the volume of the fermentation medium is 10-30%.
In some embodiments, the feed medium comprises, by weight, 20-35% brown sugar, 1-2.5% urea, and 4-5% leucine.
In some of these embodiments, the seed culture, the fermentation culture and the feed culture are at a temperature of 30-40 ℃.
The fermentation medium for improving the content of the surfactant is used for fermenting bacillus subtilis, and comprises, by weight, 4-7% of a carbon source, 0.65-2.25% of a nitrogen source, 0.13-0.6% of phosphate, 0.015-0.05% of magnesium sulfate, 0.0007-0.02% of calcium chloride, 0.0005-0.001% of manganese sulfate, 0.0006-0.06% of ferric sulfate, 0.5-1% of amino acid, 0.5-4% of soybean fibers and the balance of water, wherein the pH of the fermentation medium is 6.5-7, the inoculum size of bacillus subtilis bacterial liquid in the fermentation medium is 3-5% in terms of volume fraction, the carbon source is selected from at least one of molasses, glucose, brown sugar, maltodextrin and starch, and the nitrogen source comprises at least one of corn steep liquor dry powder, yeast paste and yeast powder in combination with urea, and the foam, so that the bacillus subtilis is capable of remarkably improving the industrial production of the surface of the bacillus subtilis, and the industrial production of the bacillus subtilis is remarkably improved.
Drawings
In order to more clearly illustrate the technical solutions of the present application or related art, the drawings that are required to be used in the description of the embodiments or related art will be briefly described below, and it is apparent that the drawings in the following description are only embodiments of the present application, and other drawings may be obtained according to the drawings without inventive effort to those of ordinary skill in the art.
FIG. 1 is a schematic flow chart of a fermentation method for increasing the content of surfactant according to an embodiment of the application.
Fig. 2 shows the difference significance results between example 1, example 2 and example 4.
Fig. 3 is a graph showing the foam suppressing effect in example 2 and example 3.
Detailed Description
The present application will be further described in detail below with reference to specific embodiments and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the present application more apparent.
It should be noted that unless otherwise defined, technical or scientific terms used in the embodiments of the present application should be given the ordinary meaning as understood by one of ordinary skill in the art to which the present application belongs. The terms "first," "second," and the like, as used in embodiments of the present application, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect.
Although the surfactant has important characteristics for wide application in industry, its high production cost and low yield in practical production limit its application. Thus, increasing the yield of surfactants and reducing the production costs remain fundamental solutions for commercial applications of surfactants. In the related art, since the production of a surfactant by fermentation of bacillus subtilis requires aeration and stirring, foam is easily produced, and thus there is a problem that foam is easily produced and the yield of the surfactant is low. The optimization of the culture medium formula is one of the effective means for improving the yield of the surfactant.
Based on the above, the embodiment of the application provides a fermentation medium and a fermentation method for improving the content of the surfactant, and by adding the soybean fiber, the yield of the surfactant produced by fermenting bacillus subtilis can be obviously improved while foam is inhibited, so that the fermentation medium is favorable for realizing industrialized mass production, and has a huge industrial application prospect. Can solve the problems of low yield and high cost of the prior surfactant to a certain extent.
The embodiment of the application provides a fermentation medium for improving the content of surfactant, which is used for fermenting bacillus subtilis, and comprises, by weight, 4-7% of a carbon source, 0.65-2.25% of a nitrogen source, 0.13-0.6% of phosphate, 0.015-0.05% of magnesium sulfate, 0.0007-0.02% of calcium chloride, 0.0005-0.001% of manganese sulfate, 0.0006-0.06% of ferric sulfate, 0.5-1% of amino acid, 0.5-4% of soybean fibers and the balance of water, wherein the pH of the fermentation medium is 6.5-7, and the inoculation amount of bacillus subtilis bacterial liquid in the fermentation medium is 3-5% in terms of volume fraction;
The carbon source is at least one selected from molasses, glucose, brown sugar, maltodextrin and starch, and the nitrogen source comprises at least one of corn steep liquor dry powder, yeast extract and yeast powder in combination with urea.
In some embodiments, the carbon source may be selected from brown sugar, and the mass percentage of the brown sugar may be 4-7%. Compared with molasses, glucose, maltodextrin and starch, the quality percentage of brown sugar can better improve the content of the surfactant obtained by fermenting bacillus subtilis.
In some embodiments, the nitrogen source is selected from the group consisting of yeast extract and urea, wherein the yeast extract is 0.45-0.55% by mass and the urea is 0.45-0.55% by mass. The yeast extract is selected, so that the content of the surfactant obtained by fermenting the bacillus subtilis can be better improved compared with corn steep liquor dry powder and yeast powder.
In some of these embodiments, the phosphate may be selected from the group consisting of potassium dihydrogen phosphate and disodium hydrogen phosphate. The mass percentage of the potassium dihydrogen phosphate may be 0.03%, and the mass percentage of the disodium hydrogen phosphate may be 0.1%.
In some of these embodiments, the amino acid may be selected from leucine. Leucine as a component of the surfactant can provide an essential component for the growth of the surfactant, thereby better increasing the content of the surfactant obtained by fermentation of bacillus subtilis.
In some embodiments, the soybean fiber can be added in an amount of 1-3%, so that the content of the surfactant obtained by fermenting the bacillus subtilis can be better improved. The addition of the soybean fiber can improve the viscosity of the fermentation medium, so that the fermentation medium is more viscous, foam generated in the fermentation process can be inhibited to a certain extent, and meanwhile, the growth of bacillus subtilis thalli and the production of product surfactant can be promoted.
Based on the same inventive concept, the embodiment of the application also provides an application of the fermentation medium for improving the surfactant content in the fermentation product of bacillus subtilis. The bacillus subtilis can be bacillus subtilis with a preservation number of CGMCC No. 24947.
Based on the same inventive concept, the embodiment of the application also provides a fermentation method for improving the content of the surfactant, as shown in fig. 1, which can include:
Step S100, inoculating bacillus subtilis into a liquid seed culture medium, and culturing the seeds for 16-18 hours to obtain fermentation strains;
Step 200, inoculating a fermentation strain into a fermentation medium, and performing fermentation culture for 22-26 hours, wherein the inoculation amount of the fermentation strain is 3-5% in terms of volume fraction, and the fermentation medium is the fermentation medium in any embodiment;
And step S300, adding a feed medium, and carrying out feed culture for 46-50h.
In some embodiments, the feed medium comprises brown sugar, urea and leucine, and the feed medium is added in an amount such that the ratio of the added volume of the feed medium to the volume of the fermentation medium is 9-11%.
In some embodiments, the feed medium comprises, by weight, 20-35% brown sugar, 1-2.5% urea, and 4-5% leucine. Generally, since the basic body of the fermentation culture is relatively viscous, the viscous state can be maintained without adding soybean fibers in a subsequent feed medium, and the fermentation culture can also have a certain capability of inhibiting foam and promoting the growth of bacillus subtilis thalli and the production of product surfactant.
In some of these embodiments, the dissolved oxygen content in the seed culture, the fermentation culture and the feed culture may be controlled to be DO >20%, and the culture temperature may be 30-40 ℃. Under the condition of shaking culture, the rotation speed of the shaking table can be 180-220r/min. The rotation speed can be 300-500rpm under the condition of fermentation tank culture.
The technical scheme of the invention is further described below with reference to the specific embodiments.
The experimental methods in the following examples are conventional methods unless otherwise specified.
The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores. In the following examples, bacillus subtilis is taken as an initial strain, and the bacillus subtilis can be bacillus subtilis with a preservation number of CGMCC No. 24947. The lipopeptide content was determined by High Performance Liquid Chromatography (HPLC) in the examples below.
Specifically, HPLC quantification may include:
(1) The chromatographic conditions are that the chromatographic column has the island fluid C18 ODS-3 (4.6 mm multiplied by 250mm,5 μm), the flow rate is 1.0mL/min, the detection wavelength is 205nm, and the column temperature is normal temperature. Mobile phase comprising acetonitrile containing 0.1% TFA and 0.1% TFA, instrument comprising Shimadzu L-033 chromatograph, gradient setting ：0～7min,A(10％～90％),B(90％～10％);7～24min,A(90％),B(10％);24～29min,A(90％～10％),B(10％～90％);29～36min,A(10％),B(90％).
(2) The measuring method comprises the steps of collecting fermentation liquor after the fermentation of bacillus subtilis in a fermentation medium is finished, adding ethanol for extraction, filtering the ethanol extraction by a filter membrane, and measuring the content by the detection method.
Example 1 Effect of Nitrogen Source on surfactant production
The test method comprises inoculating single colony of Bacillus subtilis into LB liquid medium, culturing at 37deg.C and 200rpm for 16-18h, inoculating into fermentation medium with different nitrogen sources according to inoculum size of 4v/v%, fermenting at 37deg.C and 200rpm for 24h, adding feeding medium with volume of 10% of that of the initial medium, and continuously culturing at 37deg.C and 200rpm until 48h fermentation is completed.
The basic components (namely the components of the fermentation medium) comprise 6.5 percent of maltodextrin, 0.5 percent of urea, 0.03 percent of monopotassium phosphate, 0.1 percent of disodium hydrogen phosphate, 0.015 percent of magnesium sulfate, 0.0007 percent of calcium chloride, 0.0006 percent of manganese sulfate, 0.0006 percent of ferric sulfate, 0.5 percent of leucinic acid and the pH value of 7. To the fermentation medium having the above composition, nitrogen sources such as corn steep liquor dry powder, yeast extract or yeast powder were added in an amount of 0.5%.
The formula of the feed supplement culture medium comprises 32.5% of maltodextrin, 2.5% of urea and 5% of leucine, and the pH is natural. The fermentation broth obtained after 48 hours of fermentation culture was subjected to HPLC to determine the content of the surfactant.
The experimental results are shown in Table 1 below.
TABLE 1 Effect of different kinds of Nitrogen sources on surfactant production
As a result, it was found from Table 1 that the addition of different nitrogen sources to the fermentation medium had different effects on the content of the surfactant in the fermentation product of Bacillus subtilis. Wherein, the yeast extract is added as a nitrogen source, and the obtained fermentation product of the bacillus subtilis has the highest content of the surfactant.
Example 2 Effect of carbon sources on surfactant production
The experimental method comprises inoculating single colony of Bacillus subtilis into LB liquid medium, culturing at 37deg.C and 200rpm for 16-18h, inoculating into fermentation medium with different nitrogen sources according to inoculum size of 4v/v%, fermenting at 37deg.C and 200rpm for 24h, adding feeding medium with volume of 10% of that of the initial medium, and continuously culturing at 37deg.C and 200rpm until 48h fermentation is completed.
The basic components (namely, the components of the fermentation medium) comprise 0.5% of yeast extract, 0.5% of urea, 0.03% of monopotassium phosphate, 0.1% of disodium hydrogen phosphate, 0.015% of magnesium sulfate, 0.0007% of calcium chloride, 0.0006% of manganese sulfate, 0.0006% of ferric sulfate, 0.5% of leucinic acid and pH7. 7% of one of carbon sources such as maltodextrin, glucose, brown sugar, molasses and starch is added into the above composition.
The formula of the feed supplement culture medium comprises 2.5% of urea and 5% of leucine, and the pH is natural. 35% of molasses, glucose, brown sugar, maltodextrin, starch and other carbon sources are added into the above components.
The fermentation broth obtained after 48 hours of fermentation culture was subjected to HPLC to determine the content of the surfactant.
The results of the experiment are shown in Table 2 below and FIG. 3. Wherein, the control group in fig. 3 is a foam state diagram when the carbon source is brown sugar.
TABLE 2 Effect of different kinds of carbon sources on the production of surfactant
As a result, it is clear from Table 2 that the addition of different kinds of carbon sources to the fermentation medium has different effects on the content of the surfactant in the fermentation product of Bacillus subtilis. Wherein brown sugar is added as a carbon source, and the obtained fermentation product of the bacillus subtilis has the highest content of surfactant.
Example 3 Effect of legume materials on surfactant production
The test method comprises inoculating single colony of Bacillus subtilis into LB liquid medium, culturing at 37deg.C and 200rpm for 16-18h, inoculating into fermentation medium with different nitrogen sources according to inoculum size of 4v/v%, fermenting at 37deg.C and 200rpm for 24h, adding feeding medium with volume of 10% of that of the initial medium, and continuously culturing at 37deg.C and 200rpm until 48h fermentation is completed.
The basic components (namely the components of the fermentation medium) comprise 7% of brown sugar, 0.5% of yeast extract, 0.5% of urea, 0.03% of monopotassium phosphate, 0.1% of disodium hydrogen phosphate, 0.015% of magnesium sulfate, 0.0007% of calcium chloride, 0.0006% of manganese sulfate, 0.0006% of ferric sulfate, 0.5% of leucinic acid and pH7. To the above composition, 3% of one of bean materials such as soybean meal, soybean powder, soybean fiber, etc. was added.
The formula of the feed supplement culture medium comprises 35% of brown sugar, 2.5% of urea, 5% of leucine and natural pH.
The fermentation broth obtained after 48 hours of fermentation culture was subjected to HPLC to determine the content of the surfactant.
The results of the experiment are shown in Table 3 below and FIG. 3.
TABLE 3 Effect of different classes of soybean substances on surfactant production
As a result, it was found from Table 3 that the addition of different types of soybean substances to the fermentation medium can inhibit foaming to some extent, but the effect on the content of the surfactant in the fermentation product of Bacillus subtilis is different. Wherein, after adding soybean powder and soybean fiber, only a small amount of foam was suppressed compared to the control in which the carbon source was brown sugar in example 2. In contrast, when the soybean meal was added, a large amount of foam could be suppressed as compared with the control in which the carbon source was brown sugar in example 2. The soybean meal is residue left in the oil extraction process of soybean, and mainly contains protein, fiber, mineral and other components. The soybean powder is powder prepared from soybean as raw material, and mainly contains proteins, fats, carbohydrates, minerals, dietary fibers, etc. The soybean fiber mainly refers to the general name of macromolecular saccharides which cannot be digested by digestive enzymes, and comprises cellulose, pectin, xylan, mannose and other substances. Therefore, the soybean fiber is added as a soybean substance, so that the soybean fiber has a good foam inhibition effect, and the obtained fermentation product of the bacillus subtilis has the highest content of the surfactant.
Example 4 Effect of soy fiber concentration on surfactant production
The test method comprises inoculating single colony of Bacillus subtilis into LB liquid medium, culturing at 37deg.C and 200rpm for 16-18h, inoculating into fermentation medium with different nitrogen sources according to inoculum size of 4v/v%, fermenting at 37deg.C and 200rpm for 24h, adding feeding medium with volume of 10% of that of the initial medium, and continuously culturing at 37deg.C and 200rpm until 48h fermentation is completed.
The basic components (namely fermentation medium components) comprise 7% of brown sugar, 0.5% of yeast extract, 0.5% of urea, 0.03% of monopotassium phosphate, 0.1% of disodium hydrogen phosphate, 0.015% of magnesium sulfate, 0.0007% of calcium chloride, 0.0006% of manganese sulfate, 0.0006% of ferric sulfate, 0.5% of leucinic acid and pH7. One of the soybean fibers was added to the above composition at a concentration of 0.5%, 1% and 3%.
The formula of the feed supplement culture medium comprises 35% of brown sugar, 2.5% of urea, 5% of leucine and natural pH.
The fermentation broth obtained after 48 hours of fermentation culture was subjected to HPLC to determine the content of the surfactant.
The results of the experiment are shown in Table 4 below and FIG. 2.
TABLE 4 influence of different concentrations of Soybean fibers on the surface active substances
As a result, it was found from Table 4 that the effect of adding soybean fibers at different concentrations to the fermentation medium was different in the content of the surfactant in the fermentation product of Bacillus subtilis. Wherein, soybean fiber with the concentration of 3% is added, and the obtained fermentation product of the bacillus subtilis has the highest surfactant content.
Comparing example 4 with examples 1 and 2, it is seen that the concentration of soybean fiber added was 3%, and the surfactant content in the fermentation product of Bacillus subtilis was increased by approximately 50% as compared to the nitrogen source of the yeast extract type in example 1 without soybean fiber added. The resulting fermentation product of bacillus subtilis has a nearly 30% increase in surfactant content compared to the brown sugar type carbon source of example 2 without added soybean fibers.
Fig. 2 is a graph comparing the yields under the optimal conditions in examples 1,2 and 4, and it can be seen from fig. 2 that p≤0.001 shows that the addition of the soybean fiber can increase the content of the surfactant while suppressing the foam between examples 1 and 2, examples 4 and 1, and examples 4 and 2.
EXAMPLE 5 production of surfactant in 30L fermentors
The test method comprises inoculating single colony of Bacillus subtilis into LB liquid medium at 37deg.C and 200rpm for 16-18h, inoculating into the following fermentation medium according to 4v/v% inoculum size, fermenting for 16h, feeding feed medium, continuously culturing until 48h fermentation is completed, fermenting at 37deg.C and aeration rate of 20m 2/h at rotation speed of 300-500rpm, maintaining DO >20%, and regulating and maintaining pH above 6.0 with NaOH.
The basic components (namely the components of the fermentation medium) comprise 7% of brown sugar, 0.5% of yeast extract, 0.5% of urea, 0.03% of monopotassium phosphate, 0.1% of disodium hydrogen phosphate, 0.015% of magnesium sulfate, 0.0007% of calcium chloride, 0.0006% of manganese sulfate, 0.0006% of ferric sulfate, 0.5% of leucinic acid and pH of 7. Soybean fiber 3%.
The formula of the feed supplement culture medium comprises 35% of brown sugar, 2.5% of urea, 5% of leucine and natural pH. The volume of the feed medium added was 10% of the volume of the fermentation medium.
The fermentation broth obtained after 48h of fermentation culture was determined by HPLC.
The experiment result shows that the content of the obtained surfactant is 22.04g/L.
It will be appreciated by persons skilled in the art that the foregoing discussion of any embodiment is merely exemplary and is not intended to imply that the scope of the disclosure, including the claims, is limited to these examples, that the steps may be implemented in any order and that many other variations of the different aspects of the disclosed embodiments described above are present, which are not provided in detail for the sake of brevity, and that the features of the above embodiments or of the different embodiments may also be combined within the spirit of the disclosure.
While the present disclosure has been described in conjunction with specific embodiments thereof, many alternatives, modifications, and variations of those embodiments will be apparent to those skilled in the art in light of the foregoing description.
The disclosed embodiments are intended to embrace all such alternatives, modifications and variances which fall within the broad scope of the appended claims. Accordingly, any omissions, modifications, equivalents, improvements, and the like, which are within the spirit and principles of the embodiments of the disclosure, are intended to be included within the scope of the disclosure.
Claims (10)
1. A fermentation medium for improving the content of surface active substances is characterized by being used for fermenting bacillus subtilis (Bacillus subtilis), and comprises, by weight, 4-7% of a carbon source, 0.65-2.25% of a nitrogen source, 0.13-0.6% of phosphate, 0.015-0.05% of magnesium sulfate, 0.0007-0.02% of calcium chloride, 0.0005-0.001% of manganese sulfate, 0.0006-0.06% of ferric sulfate, 0.5-1% of amino acid, 0.5-3% of soybean fibers and the balance of water, wherein the pH of the fermentation medium is 6.5-7, and the inoculation amount of bacillus subtilis bacterial liquid in the fermentation medium is 3-5% in terms of volume fraction;
The carbon source is at least one selected from molasses, glucose, brown sugar, maltodextrin and starch, and the nitrogen source comprises at least one of corn steep liquor dry powder, yeast extract and yeast powder in combination with urea.
2. The fermentation medium for increasing a surfactant content according to claim 1, wherein the soybean fiber is added in an amount of 1 to 3%.
3. The fermentation medium for increasing the surfactant content according to claim 2, wherein the nitrogen source is selected from the group consisting of yeast extract and urea, the mass percentage of the yeast extract is 0.45-0.55%, and the mass percentage of the urea is 0.45-0.55%.
4. The fermentation medium for increasing a surfactant content according to claim 1, wherein the carbon source is selected from brown sugar, and the mass percentage of the brown sugar is 4-7%.
5. The fermentation medium of claim 1, wherein the phosphate is selected from the group consisting of potassium dihydrogen phosphate and disodium hydrogen phosphate, and the amino acid is selected from the group consisting of leucine.
6. Use of a fermentation medium according to any one of claims 1 to 5 for increasing the surfactant content in a fermentation product of bacillus subtilis.
7. A fermentation process for increasing the level of surfactant comprising:
inoculating bacillus subtilis into a liquid seed culture medium, and carrying out seed culture for 16-18h to obtain fermentation strains;
Inoculating a fermentation strain into a fermentation culture medium for fermentation culture for 22-26h, wherein the inoculation amount of the fermentation strain is 3-5% in terms of volume fraction, and the fermentation culture medium is the fermentation culture medium according to any one of claims 1-6;
And adding a feed medium, and carrying out feed culture for 46-50h.
8. The method for increasing a surfactant content according to claim 7, wherein the feed medium comprises brown sugar, urea and leucine, and the feed medium is added in an amount such that the ratio of the volume of the feed medium to the volume of the fermentation medium is 9-11%.
9. The method for increasing a surfactant content according to claim 8, wherein the feed medium comprises, by weight, 20-35% of brown sugar, 1-2.5% of urea and 4-5% of leucine.
10. The fermentation process for increasing surfactant content according to claim 7, wherein the seed culture, the fermentation culture and the feed culture are conducted at a temperature of 30-40 ℃.
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