Pediococcus pentosaceus with cholesterol-lowering and antibacterial effects and high-density industrial production fermentation medium thereof

Abstract

The application discloses a Pediococcus pentosaceus with the functions of lowering cholesterol and inhibiting bacteria and a high-density industrial production fermentation medium of the Pediococcus pentosaceus, the Pediococcus pentosaceus is Pediococcus pentosaceus PG-2, and the preservation number is GDMCC No: 62392.The high-density industrial fermentation medium of the Pediococcus pentosaceus PG-2 contains 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, 1.5 g / L of dipotassium hydrogen phosphate, 5 g / L of sodium acetate, 1 mL / L of Tween 80, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate heptahydrate, 0.05 g / L of manganese sulfate tetrahydrate, and the rest is water.The Pediococcus pentosaceus is cultured by using the medium, the bacterial concentration is significantly higher than that of MRS medium, a high-density bacterial body amount is effectively obtained, the utilization rate of industrial by-products is improved, the production cost is greatly reduced, and the production efficiency is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, in particular to a Pediococcus pentosaceus with the functions of reducing cholesterol and inhibiting bacteria and a high-density industrial production fermentation medium thereof. BACKGROUND

[0002] Hyperlipidemia is a metabolic disease caused by the disruption of lipid metabolism balance in the body, resulting in higher blood lipid than the average level. According to the investigation of the World Health Organization, the main factor of the onset of diseases such as atherosclerosis and coronary heart disease is hyperlipidemia. Related reports indicate that if the cholesterol level in the body is higher than the average level of 1 mmol / L, there is a 35% probability of suffering from coronary heart disease. Data shows that the mortality rate caused by hyperlipidemia and related diseases accounts for one half of the mortality rate in the world.

[0003] The main method to regulate the cholesterol level in the body is to prevent the endogenous cholesterol from synthesizing itself, to reduce the absorption of exogenous cholesterol through diet adjustment, and to promote the excretion of cholesterol. In the 1970s and 1980s, foreign countries began to study the effect of probiotics on reducing blood lipids. Two strains of bacteria with deep research on the function of reducing blood lipids in China belong to Lactobacillus plantarum, which are L. plantarum P8 and ST-III, respectively. Animal experiments show that both L. plantarum P8 and ST-III have significant effects on reducing blood lipids. Therefore, the development of safe and effective lactic acid bacteria for regulating blood lipid balance has broad prospects in the fields of food and medicine. Pediococcus pentosaceus is an inherent strain of animal intestinal microorganisms, which can inhibit the proliferation of pathogenic bacteria and improve the body's disease resistance and immune function by itself. However, the existing production process produces Pediococcus pentosaceus with low viable count and high production cost, which directly affects its use effect and application promotion. Therefore, the fermentation production of Pediococcus pentosaceus needs to be further developed. SUMMARY

[0004] The present application aims to solve the problems of low viable count and high production cost of Pediococcus pentosaceus in the prior art and alleviate the severe situation of the increasing number of hyperlipidemia population in China. The present application provides a Pediococcus pentosaceus with the functions of reducing cholesterol and inhibiting bacteria and a high-density industrial production fermentation medium thereof. By using the medium, a high-density bacterial mass can be obtained, and the viable count after fermentation is significantly higher than that of the MRS medium, which improves the viable count of Pediococcus pentosaceus and reduces the production and fermentation cost.

[0005] The first object of the present application is to provide a Pediococcus pentosaceus PG-2, which has a preservation number of GDMCC No: 62392.

[0006] The Pediococcus pentosaceus PG-2 is derived from the feces of Asian small-clawed otter. The colony is milky white, smooth, with neat edges and opaque under naked eye observation. The colony is purple under Gram staining, and is gram-positive. The colony is round under microscope observation. The 16S rRNA sequence of the Pediococcus pentosaceus PG-2 is 1356 bp in length, and the specific sequence is shown as SEQ ID NO. 1. The Pediococcus pentosaceus PG-2 is identified as Pediococcus pentosaceus, and is named as Pediococcus pentosaceus PG-2.

[0007] The second object of the present application is to provide a preparation containing the Pediococcus pentosaceus PG-2 or the fermentation liquor thereof as an active ingredient.

[0008] The third object of the present application is to provide the Pediococcus pentosaceus PG-2 or the preparation as described above in at least one of the following (1)-(4):

[0009] (1) preventing and treating pathogenic bacteria;

[0010] (2) preparing a drug or health product for reducing cholesterol;

[0011] (3) preparing animal feed or feed additive;

[0012] (4) preparing food or food additive.

[0013] Preferably, the pathogenic bacteria include Staphylococcus aureus, Escherichia coli and Salmonella enteritidis.

[0014] The fourth object of the present application is to provide a high-density industrial production fermentation medium of Pediococcus pentosaceus, which contains 30-40 g / L of sugarcane molasses, 20-25 g / L of corn syrup dry powder, 1.5 g / L of dipotassium hydrogen phosphate, 5 g / L of sodium acetate, 1 mL / L of Tween 80, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate heptahydrate, 0.05 g / L of manganese sulfate tetrahydrate, and water as a solvent.

[0015] Preferably, the fermentation medium contains 40 g / L of sugarcane molasses, 20 g / L of corn syrup dry powder, 1.5 g / L of dipotassium hydrogen phosphate, 5 g / L of sodium acetate, 1 mL / L of Tween 80, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate heptahydrate, 0.05 g / L of manganese sulfate tetrahydrate, and water as a solvent.

[0016] The fifth object of the present application is to provide the use of the high-density industrial production fermentation medium as described above in the high-efficiency fermentation of Pediococcus pentosaceus.

[0017] A sixth object of the present application is to provide a method for efficiently fermenting Pediococcus pentosaceus, which inoculates the above-mentioned Pediococcus pentosaceus PG-2 into the above-mentioned high-density industrial production fermentation medium to perform fermentation culture.

[0018] Preferably, the method comprises the following steps: activating the preserved Pediococcus pentosaceus PG-2 on an MRS agar medium plate, inoculating a single colony into an MRS liquid medium to perform culture, and then transferring the bacterial liquid into a high-density industrial production fermentation medium to perform fermentation culture.

[0019] Preferably, the inoculation of the single colony into the MRS liquid medium to perform culture is performed under the following conditions: 37℃ for 14 hours; the transferring is performed by transferring the bacterial liquid at a volume fraction of 1%; and the fermentation culture is performed under the following conditions: 37℃ for 48 hours.

[0020] Compared with the prior art, the present application has the following beneficial effects:

[0021] (1) The Pediococcus pentosaceus PG-2 of the present application can produce antibacterial and bacteriostatic substances, has good bacteriostatic effect on common livestock and poultry and aquatic breeding pathogenic bacteria, including good inhibition effect on Staphylococcus aureus, Escherichia coli and Salmonella enteritidis, and also has excellent cholesterol-lowering effect, with a cholesterol removal rate as high as (40.66±0.25) %;

[0022] (2) The Pediococcus pentosaceus PG-2 high-density industrial production fermentation medium of the present application is suitable for large-scale fermentation of Pediococcus pentosaceus. The industrial by-product sugarcane molasses is used as a carbon source, and corn syrup dry powder is used as a main nitrogen source to culture Pediococcus pentosaceus, thereby reducing the production cost and being beneficial to the rapid growth of Pediococcus pentosaceus, and the number of viable bacteria after fermentation is significantly improved compared with MRS medium.

[0023] The Pediococcus pentosaceus PG-2 of the present application was preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on April 18, 2022, at address No. 59, Building 5, 100, Martyrs' Road, Guangzhou, with postcode 510070, and preservation number GDMCC No: 62392. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 It is a colony morphology diagram of the Pediococcus pentosaceus PG-2 of the present application.

[0025] Figure 2 It is a morphology diagram of the Pediococcus pentosaceus PG-2 of the present application under a microscope.

[0026] Figure 3Columnar diagram and bacteriostatic picture of the supernatant of the fermentation liquor of Pediococcus pentosaceus PG-2 after centrifugation on the inhibition effect on three pathogenic bacteria.

[0027] Figure 4 Effect of single inorganic salt on the viable count of Pediococcus pentosaceus PG-2.

[0028] Figure 5 Effect of multiple inorganic salts on the viable count of Pediococcus pentosaceus PG-2. DETAILED DESCRIPTION

[0029] The following is a further elaboration of the content of the present application combined with implementation cases, but is not a limitation of the present application. In the following examples, all consumables, instruments and related supplies and detection methods are conventional unless otherwise specified.

[0030] Example 1: Isolation, purification, identification and preservation of Pediococcus pentosaceus PG-2

[0031] (1) Isolation, screening and preservation of Pediococcus pentosaceus PG-2

[0032] The Pediococcus pentosaceus PG-2 of the present application is isolated from the feces of Asian small-clawed otter.

[0033] MRS agar medium: 10 g of casein enzymatic digest, 10 g of beef extract powder, 4 g of yeast extract powder, 20 g of glucose, 2 g of potassium phosphate dibasic, 2 g of triammonium citrate, 5 g of sodium acetate, 0.2 g of magnesium sulfate heptahydrate, 0.05 g of manganese sulfate tetrahydrate, 1.08 g of Tween-80, 15 g of agar, mix the above ingredients, add water to 1000 mL, adjust pH to 5.7±0.2, divide into 250 mL triangular bottles, 100 mL per bottle; sterilize at 121℃ high temperature and high pressure for 20 min, pour flat plate.

[0034] The feces of Asian small-clawed otter is coated on the plate containing MRS agar medium by dilution coating method, and strain PG-2 is obtained after purification. The colony morphology is observed by naked eye, and the results are shown in Figure 1 . The colony is milky white, smooth in surface, regular in edge, and opaque; gram staining is purple, which is gram-positive bacteria; the bacterial shape is observed under a microscope Figure 2 ).

[0035] The genomic DNA of the strain PG-2 was extracted, and a PCR product was obtained by amplification with bacterial 16S rRNA gene universal primers 27F / 1492R (27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-TACGACTTAACCCCAATCGC-3'), and was sent to Shanghai Meiji Biomedicine Technology Co., Ltd. (Guangzhou Branch) for sequence sequencing. The 16S rRNA sequence obtained after sequencing has a length of 1356 bp, and the specific sequence is shown as SEQ ID NO. 1. The sequencing results were subjected to homology comparison analysis with the 16S rRNA sequences in the EzBioCloud database, combined with morphological observation, and the isolated strain PG-2 was preliminarily identified as Pediococcus pentosaceus. It was named: Pediococcus pentosaceus PG-2. The strain was preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on April 18, 2022, the address is: No. 59 Building, 5th Floor, Institute of Martyrs, Guangzhou, Guangdong Province, China, postcode: 510070, and the preservation number is GDMCC No: 62392.

[0036] The 16S rRNA sequence of Pediococcus pentosaceus PG-2 (SEQ ID NO. 1) is specifically as follows:

[0037]

[0038] Example 2: Pediococcus pentosaceus PG-2 bacteriostatic effect determination

[0039] (1) Preparation of test bacteria supernatant: The activated Pediococcus pentosaceus PG-2 was inoculated into MRS liquid medium (except that no agar was added, the formula was the same as MRS agar medium) at a volume fraction of 5%, and cultured in a 37℃ three-gas incubator for 18h, the oxygen content in the three-gas incubator was 5%, and the carbon dioxide content was 10%; after centrifugation, the supernatant was taken and stored at 4℃.

[0040] (2) Preparation of bacteria-containing plates: The test strain was inoculated into nutrient broth medium (formula: proteose peptone 10.0g / L, beef extract powder 3.0g / L, sodium chloride 5.0g / L, pH 7.4±0.2, solvent water) and cultured in a 30℃ constant temperature incubator (as shown in Table 1), and the OD value was adjusted to 0.1 by ultraviolet spectrophotometer, then 0.1mL of bacterial solution was taken and added to the corresponding 100mL medium melted at about 45℃, and then poured into plates after shaking, and then the plates were naturally solidified, and then punched with a puncher to make holes (hole diameter 9mm) on the same plate. 600

[0041] (3) Bacteriostatic test

[0042] 100μL of Pediococcus pentosaceus PG-2 supernatant after centrifugation was added to each hole of the plate containing the test pathogenic bacteria, and each group had three repeats, and then cultured in a 30℃ incubator for about 24h, then the bacteriostatic effect was observed, and the diameter of the bacteriostatic ring was measured (results shown in Figure 3 ).

[0043] Table 1. Test strains for Pediococcus pentosaceus PG-2 bacteriostatic effect test

[0044]

[0045] From Figure 3 it can be seen that the supernatant of Pediococcus pentosaceus PG-2 has good inhibitory effect on Staphylococcus aureus, Escherichia coli and Salmonella enteritidis.

[0046] Example 3: Determination of BSH enzyme activity of Pediococcus pentosaceus PG-2

[0047] (1) Preparation of taurine standard curve

[0048] The standard curve is prepared by taking the concentration (μmol / mL) of taurine as the abscissa and the absorbance value (A570nm) as the ordinate, and the standard curve of taurine is y=0.2286x-0.0041, R 2 =0.9923. ​

[0049] (2) Preparation and determination of samples

[0050] After the Pediococcus pentosaceus PG-2 of the application was activated for 3 times, it was inoculated into MRS liquid medium at a volume fraction of 3%, and cultured at 37°C for 24 hours. Then, a proper amount of fermentation liquor was centrifuged to collect the bacterial cells. The bacterial cells were washed twice with 0.1 mol / L (pH 7.0) phosphate buffer, and the absorbance thereof was adjusted to 1.0 at 600 nm. Then, dithiothreitol was added to prevent the oxidation of bile salt hydrolase (BSH). 1 mL of the adjusted bacterial liquor was ultrasonically broken, and centrifuged to obtain a cell-free extract. 10 μL of the cell-free extract was mixed with 180 μL of 0.1 mol / L phosphate buffer, 10 μL of 200 mmol / L sodium taurocholate and 10 μL of paraffin oil, and then placed at 37°C for 30 minutes. Then, the same volume of trichloroacetic acid was added to terminate the reaction, and the mixture was centrifuged to obtain the supernatant. 0.1 mL of the supernatant was mixed with 1.9 mL of ninhydrin color developing solution, and then heated in boiling water for 14 minutes. After cooling with tap water, the absorbance of the mixture was determined at 570 nm by using an enzyme label meter. The positive control strain was Lactobacillus plantarum GDMCC 1.140.

[0051] Definition of the crude enzyme activity of BSH: the amount of substance of amino acids produced by the hydrolysis of conjugated bile salts per unit time and per unit volume, and the unit is μmol / (h·mL). BSH can hydrolyze conjugated bile salts to produce amino acids and free bile acids, and the free bile acids can form a complex with cholesterol to be discharged out of the body, thereby reducing the content of cholesterol in the blood. Therefore, BSH can be used as an important indicator for screening strains capable of degrading cholesterol.

[0052] The results show that the crude enzyme activity of the BSH-producing Pediococcus pentosaceus PG-2 is as high as 0.75 μmol / (h·mL), and the crude enzyme activity of the positive control Lactobacillus plantarum GDMCC 1.140 is 0.70 μmol / (h·mL), indicating that the Pediococcus pentosaceus PG-2 has the potential for high-efficiency cholesterol reduction.

[0053] Example 4: Determination of the cholesterol removal effect of Pediococcus pentosaceus PG-2

[0054] (1) Preparation of a standard curve of cholesterol

[0055] The standard curve of cholesterol was prepared by taking the concentration of cholesterol (mg / mL) as the horizontal coordinate and the absorbance (A500 nm) as the vertical coordinate, and the standard curve of cholesterol was y=24.393x-0.0125, R 2 = 0.9944.

[0056] (2) Preparation and determination of samples

[0057] After the Pediococcus pentosaceus PG-2 of the application is activated for 3 times, the Pediococcus pentosaceus PG-2 is inoculated into a MRS medium containing cholesterol (MRS-CHOL medium) at a volume fraction of 3%, and is cultured at 37°C for 24 hours, 1 mL of fermentation liquor is centrifuged to obtain a supernatant, and then the content of cholesterol in the supernatant is determined by using an o-phthaldehyde method, and the removal rate of cholesterol of different strains is calculated according to the following formula. The MRS-CHOL medium is a MRS liquid medium to which 3% cholate and 1% cholesterol are added. The positive control strain is Lactobacillus plantarum GDMCC1.140.

[0058]

[0059] In the formula, A is the concentration of cholesterol in the medium before inoculation; and B is the concentration of cholesterol in the medium after inoculation and fermentation.

[0060] The results show that the cholesterol removal rate of the Pediococcus pentosaceus PG-2 is as high as (40.66±0.25)%, and the cholesterol removal rate of the positive control Lactobacillus plantarum GDMCC1.140 is 45%, which indicates that the Pediococcus pentosaceus PG-2 has a high cholesterol-lowering capacity. The strain PG-2 of the application can be applied to products with a cholesterol-degrading function, such as feed additives.

[0061] Example 5: Comparison of fermentation single-factor test results of inorganic salts in the medium

[0062] The composition of each mass component of the fermentation medium is as follows:

[0063] Medium 1: MRS liquid medium;

[0064] Medium 2: 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, 1.5 g / L of potassium phosphate dibasic, 5 g / L of sodium acetate, 1 mL / L of Tween80, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate heptahydrate, 0.05 g / L of manganese sulfate tetrahydrate, and water as a solvent;

[0065] Medium 3: 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, and water as a solvent;

[0066] Medium 4: 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, 1.5 g / L of potassium phosphate dibasic, and water as a solvent;

[0067] Medium 5: 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, 5 g / L of sodium acetate, and water as a solvent;

[0068] Culture medium 6: 40 g / L sugarcane molasses solution, 20 g / L corn steep liquor powder, 1 mL / L Tween 80, with water as the solvent;

[0069] Culture medium 7: 40 g / L sugarcane molasses solution, 20 g / L corn steep liquor powder, 2 g / L triammonium citrate, and water as solvent;

[0070] Culture medium 8: 40 g / L sugarcane molasses solution, 20 g / L corn steep liquor powder, 0.2 g / L magnesium sulfate heptahydrate, with water as the solvent;

[0071] Culture medium 9: 40 g / L sugarcane molasses solution, 20 g / L corn steep liquor powder, 0.05 g / L manganese sulfate tetrahydrate, with water as the solvent;

[0072] The pH of all the above culture media is 6.0.

[0073] The preparation method of the above culture medium is as follows: mix the corresponding components of the above culture medium evenly, dissolve them with distilled water, put the prepared culture medium into a 250mL Erlenmeyer flask, stir evenly, and sterilize at 115℃ for 20min.

[0074] To achieve the best results from the culture medium, the fermentation process for *Pediococcus pentosaceus* using the above-mentioned medium is as follows:

[0075] (1) Activate the frozen Pediococcus pentosacchari PG-2 on an MRS agar plate (the composition and preparation method are the same as in Example 1), pick a single colony and inoculate it into a test tube containing MRS liquid medium, and incubate at 37°C for about 14 hours until the liquid medium becomes obviously turbid to obtain the seed culture solution.

[0076] (2) According to the culture medium formula, prepare 50 mL of each of the above 9 fermentation culture media and put them into 250 mL Erlenmeyer flasks. After sterilization, lower the temperature to 37℃.

[0077] (3) The seed culture solution obtained was inoculated into Erlenmeyer flasks containing different fermentation media at an inoculation amount of 1% by volume; 3 replicates per group.

[0078] (4) The fermentation conditions are set as follows: temperature 37℃, fermentation time 48 hours, and the fermentation liquid after fermentation is completed is diluted and coated.

[0079] The viable cell counts of the bacterial cultures obtained using the above nine fermentation media are shown in the figure. Figure 4 . Figure 4 The results showed that *Pediococcus pentosaceus* PG-2 had the highest viable count in medium 2, followed by medium 1 and medium 7. Since the viable count in medium 7 was significantly higher than that in basal medium 3, medium 7 was selected as the optimized fermentation medium.

[0080] Example 6: Comparison of the results of the inorganic salt fermentation factor test in the medium

[0081] The composition of each mass component of the fermentation medium is as follows:

[0082] Medium 1: MRS liquid medium

[0083] Medium 2: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, dipotassium hydrogen phosphate 1.5 g / L, sodium acetate 5 g / L, Tween 80 1 mL / L, triammonium citrate 2 g / L, magnesium sulfate heptahydrate 0.2 g / L, manganese sulfate tetrahydrate 0.05 g / L, solvent is water

[0084] Medium 3: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, solvent is water

[0085] Medium 4: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, triammonium citrate 2 g / L, dipotassium hydrogen phosphate 1.5 g / L, solvent is water

[0086] Medium 5: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, triammonium citrate 2 g / L, Tween 80 1 mL / L, solvent is water

[0087] Medium 6: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, triammonium citrate 2 g / L, magnesium sulfate heptahydrate 0.2 g / L, solvent is water

[0088] Medium 7: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, triammonium citrate 2 g / L, manganese sulfate tetrahydrate 0.05 g / L, solvent is water

[0089] Medium 8: Sugarcane molasses liquid 40 g / L, corn syrup dry powder 20 g / L, triammonium citrate 2 g / L, solvent is water

[0090] The pH of the above-mentioned medium is 6.0. Each group has 3 repeats.

[0091] The medium preparation method and the fermentation process are the same as in Example 5.

[0092] The viable cell count of the bacterial liquid obtained by using the above-mentioned 8 kinds of fermentation medium for fermentation culture is shown in Table 2 Figure 5 . Figure 5The results show that the number of viable cells of P. pentosaceus PG-2 in medium 2 is the highest, followed by medium 1 and medium 4. The number of viable cells in medium 1 and medium 2 is significantly higher than that in medium 4, and the number of viable cells in medium 4 and medium 8 is not significantly different, so medium 2 is preferred as the industrial high-density fermentation medium of P. pentosaceus PG-2, which is helpful for the growth of P. pentosaceus PG-2. And the carbon source and nitrogen source of this medium use industrial by-products, so the fermentation cost is significantly lower than that of MRS liquid medium.

[0093] The above is only the preferred embodiment of the present application, it should be noted that the above preferred embodiment should not be considered as a limitation of the present application, the protection scope of the present application should be limited by the scope defined by the claims. For ordinary skilled in the art, several improvements and refinements can be made without departing from the spirit and scope of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A strain of Pediococcus pentosaceus ( Pediococcus pentosaceus PG-2, with accession number GDMCC No: 62392.

2. A formulation characterized in that, Pediococcus pentosaceus containing the Pediococcus pentosaceus of claim 1 Pediococcus pentosaceus ) PG-2 as an active ingredient.

3. The P. pentosus of claim 1 (P. pentosus (P. cerevisiae)) PG-2 or the preparation of claim 2 for use in at least one of the following (1)-(2): Pediococcus pentosaceus ) PG-2 or the preparation of claim 2 for use in at least one of the following (1)-(2): (1) preparing the medicine for reducing cholesterol or the health care product for assisting reducing cholesterol; (2) preparing food or food additive.

4. Use of a high-density industrial production fermentation medium in the efficient fermentation of Pediococcus pentosaceus, characterized in that, The Pediococcus pentosaceus described in claim 1 ( Pediococcus pentosaceus PG-2 was inoculated into the high-density industrial fermentation medium for fermentation culture. The high-density industrial fermentation medium contained 30-40 g / L sugarcane molasses, 20-25 g / L corn steep liquor powder, 1.5 g / L dipotassium hydrogen phosphate, 5 g / L sodium acetate, 1 mL / L Tween 80, 2 g / L triammonium citrate, 0.2 g / L magnesium sulfate heptahydrate, 0.05 g / L manganese sulfate tetrahydrate, and water as the solvent.

5. Use according to claim 4, characterized in that, The high-density industrial production fermentation medium contains 40 g / L of sugarcane molasses liquid, 20 g / L of corn syrup dry powder, 1.5 g / L of potassium phosphate dibasic, 5 g / L of sodium acetate, 1 mL / L of Tween 80, 2 g / L of triammonium citrate, 0.2 g / L of magnesium sulfate heptahydrate, 0.05 g / L of manganese sulfate tetrahydrate, and water as a solvent.

6. Use according to claim 4, characterized in that, Preserved Pediococcus pentosaceus ( Pediococcus pentosaceus PG-2 was activated on MRS agar plates, and single colonies were picked and inoculated into MRS liquid medium for culture. The bacterial culture was then transferred to high-density industrial fermentation medium for fermentation culture.

7. Use according to claim 6, characterized in that, The single colony is inoculated in MRS liquid medium for culture, and the culture condition is 37℃ for 14 hours; the transfer is carried out by inoculating the bacterial liquid with a volume fraction of 1%; and the fermentation culture is carried out under the culture condition of 37℃ for 48 hours.

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