CN113930367A

CN113930367A - Lactic acid bacteria with cholesterol-reducing performance and application thereof

Info

Publication number: CN113930367A
Application number: CN202111342225.1A
Authority: CN
Inventors: 刘玲彦; 杜维力; 李啸; 谈亚丽; 白玛卓嘎; 王庆宇
Original assignee: Hubei Anqi Biology Group Co ltd
Current assignee: Angel Biotechnology Co ltd
Priority date: 2021-11-12
Filing date: 2021-11-12
Publication date: 2022-01-14
Anticipated expiration: 2041-11-12
Also published as: CN113930367B

Abstract

The invention provides a lactic acid bacterium with cholesterol-lowering performance and application thereof. The Weissella cibaria strain provided by the invention is a Weissella cibaria LMLC4560-1 strain (Weissella cibaria LMLC4560-1), which is preserved in China Center for Type Culture Collection (CCTCC) at 7 months and 2 days in 2021, and the preservation number is CCTCC NO: M2021811. The Weissella cibaria LMLC4560-1 strain provided by the invention has high cholesterol-lowering capability and strong antioxidant capability, and has good gastrointestinal adverse environment resistance.

Description

Lactic acid bacteria with cholesterol-reducing performance and application thereof

Technical Field

The invention belongs to the field of microorganisms, and particularly relates to a lactic acid bacterium with cholesterol-lowering performance and application thereof.

Background

Weissella belongs to lactic acid bacteria, is widely distributed, contains various Weissella in the environments of food, soil, gastrointestinal tract, respiratory tract and the like, and the fermented food is a rich source of Weissella. Some Weissella derived from fermented foods can significantly reduce biogenic amines in fermented foods due to the fact that the genome contains the sufI gene encoding copper oxidase. The Weissella metabolizes to produce bacteriocin, which can prevent the growth of spoilage organisms and prolong the shelf life of food. Weissella also has the ability to synthesize oligosaccharides, and the product can be used as flavoring agent in food for diabetes patients and obesity patients, and can also be used in cosmetic and feed industries. Research shows that Weissella cibaria has the activities of resisting cancer, regulating immunity, inhibiting inflammation level and resisting oxidation.

Hypercholesterolemia in humans has been shown to be the leading cause of atherosclerosis, leading to cardiovascular and cerebrovascular disease. High cholesterol is associated with a number of metabolic syndromes, including obesity, hyperlipidemia, hyperglycemia, hypertension, and the like. When the concentration of total cholesterol in serum of a human body exceeds 7.28mmol/L, the risk degree of acute coronary heart disease is also greatly increased. Therefore, lowering the intake of cholesterol through diet suppression and developing foods with the function of regulating the cholesterol content in the body are healthier and more effective ways, and probiotics with the cholesterol-lowering efficacy are promising functional ingredients.

At present, cholesterol is reduced mainly by means of drug therapy, and statin drugs can reduce serum cholesterol and cause adverse reactions such as muscle spasm and rhabdomyolysis of the body. The other main mode is diet therapy, and although certain effect is achieved, the effect is greatly weakened along with the prolonging of time, and meanwhile, long-term diet therapy is difficult to insist on. Therefore, functional foods can be produced at once without side effects and long-term persistence, and people have higher and higher attention to the products, which not only have high nutritive value per se, but also meet the public health requirements and the development requirements of the food industry. Therefore, the research and the utilization of the probiotics for reducing the cholesterol for developing the functional food and the microecological preparation for reducing the cholesterol have important significance for improving the health level of people.

Disclosure of Invention

The invention aims to provide a Weissella cibaria strain with high cholesterol-reducing capability.

The Weissella cibaria strain provided by the invention also has stronger oxidation resistance and good gastrointestinal tract adverse environment resistance.

The Weissella cibaria strain provided by the invention is separated from Tibet yak milk and has food properties.

The invention provides a Weissella sinus strain, which is a Weissella sinus LMLC4560-1 strain (Weissella cibaria LMLC4560-1), and the Weissella sinus LMLC4560-1 strain (Weissella cibaria LMLC4560-1) is preserved in China Center for Type Culture Collection (CCTCC) at 7 and 2 days 2021, and the preservation number is CCTCC NO: M2021811.

Preferably, the 16S rDNA gene sequence of the Weissella sinus LMLC4560-1 strain is shown in SEQ ID No. 2.

The invention also provides a fermentation preparation method of the Weissella cibaria microbial inoculum, which comprises the following steps: culturing the Weissella cibaria strain.

Preferably, the preparation method comprises the following steps:

(1) amplifying and culturing the Weissella cibaria strain;

(2) adding the product obtained in the step (1) into a liquid culture medium, and fermenting and culturing at the temperature of 26-32 ℃.

The invention also provides a microbial inoculum which is obtained by the preparation method.

The invention also provides the Weissella cibaria strain or a fermentation product obtained by fermenting the microbial inoculum.

Preferably, the fermentation product has cholesterol lowering properties;

preferably, the fermentation product has free radical scavenging ability.

Preferably, the fermentation product acts on cholesterol with a cholesterol reduction rate of more than 87%;

alternatively, preferably, the fermentation product has a DPPH.radical clearance of greater than 90%.

The invention also provides a preparation method of the fermentation product, and the preparation method comprises the step of culturing the Weissella cibaria strain or the microbial inoculum.

Preferably, the preparation method comprises the following steps:

(1) amplifying and culturing the Weissella cibaria strain or the microbial inoculum;

(2) adding the product obtained in the step (1) into a liquid culture medium, and fermenting and culturing at 26-32 ℃ to obtain the fermentation product.

The invention also provides application of the Weissella cibaria strain or the microbial inoculum in products for reducing cholesterol, lowering lipid, losing weight, resisting oxidation or resisting aging.

Preferably, the product is a food, cosmetic, nutraceutical, or pharmaceutical.

The Weissella cibaria LMLC4560-1 strain provided by the invention belongs to lactic acid bacteria, is derived from yak milk of herdsmen in Tibet regions, and has food properties. The Weissella cibaria LMLC4560-1 strain provided by the invention has high cholesterol-lowering capability and strong oxidation resistance, has good gastrointestinal adverse environment resistance, and supplements the strain resources of probiotics with the cholesterol-lowering function. The Weissella cibaria fermented food provided by the invention can enrich the nutritional ingredients of the food and also has the effects of improving the palatability, digestibility and the like of the food. After the food raw materials are fermented by the Weissella cibaria provided by the invention, proteins, fats and sugars can be decomposed into a predigestion state which is easier to absorb by human bodies, and simultaneously, the contents of soluble calcium, phosphorus, iron and certain B vitamins can be increased, and the digestion and absorption performance and the nutritional value are improved.

Information on strain preservation

The Weissella cibaria strain provided by the invention is a Weissella cibaria LMLC4560-1 strain (Weissella cibaria LMLC4560-1) which is preserved in China Center for Type Culture Collection (CCTCC) at 7 months and 2 days in 2021, wherein the preservation number is CCTCC NO: M2021811, the preservation address is as follows: china, wuhan university, zip code: 430072; telephone: 027-68754052.

The Lactobacillus plantarum LP-23(Lactobacillus plantarum LP-23) in the embodiment of the invention is preserved in China Center for Type Culture Collection (CCTCC) in 6.2.2020, with the preservation number of CCTCC NO: M2020175, and the preservation address: china, wuhan university, zip code: 430072; telephone: 027-68754052.

Drawings

FIG. 1 shows the standard curve for cholesterol as described in example 3;

FIG. 2 shows a colony morphology of Weissella sinus LMLC4560-1 strain;

FIG. 3 shows the morphological feature diagram of Weissella sinus LMLC4560-1 strain.

Detailed Description

The Weissella cibaria LMLC4560-1 strain provided by the invention is separated from Sublin Yak milk in Calchad county in south of Tibet. The Weissella cibaria LMLC4560-1 strain provided by the invention is a gram-positive strain with acid production performance obtained by screening strains enriched in Tibet yak milk through cycloheximide and colistin sulfate. The Weissella sinus LMLC4560-1 strain provided by the invention has food attributes, has high-efficiency cholesterol-lowering capability and strong antioxidant capability, and has good gastrointestinal adverse environment resistance. The product can be used for fermented food, and has effects of enriching nutrient components of food, and improving palatability and digestibility of food.

The specific sources of reagents used in the examples are listed in table 1 below.

TABLE 1 reagent and Instrument information for use in the present invention

Reagent/instrument	Purity/type	Manufacturer of the product
			Yeast extract powder	99％	Angel Yeast Co.,Ltd.
Glucose	99％	Yishuidi pharmaceutical Co Ltd
			Litmus milk culture medium	BR	Beijing Solaibao Biotech Co., Ltd
MRS broth	BR	Beijing Oobo Star Biotechnology Ltd
			BL agar medium	BR	Beijing Solaibao Biotech Co., Ltd
Gram staining solution	99％	Beijing Solaibao Biotech Co., Ltd
			Super PCR Mix	99.8％	BEIJING TSINGKE XINYE BIOLOGICAL TECHNOLOGY Co.,Ltd.
Cycloheximide	＞94％	Hunan Hezhongzhou Biotechnology Ltd
			Colistin sulfate	BR	Hunan Hezhongzhou Biotechnology Ltd
Cholesterol	AR	Sigma-Aldrich
			Ortho-phthalaldehyde	AR	Meclines reagent net
Ultraviolet spectrophotometer	SP-754	SHANGHAI SPECTRUM INSTRUMENTS Co.,Ltd.
			High-speed centrifugal machine	Minispin	eppendorf
PCR instrument	C1000Touch	Bio-rad
			Biochemical incubator	SPX250BSH-II	Shanghai CIMO Medical Instrument Co.,Ltd.

EXAMPLE 1 enrichment, isolation and purification of gram-Positive bacilli

Enriching strains: taking 1ml of Sungshan southern city Gatchahi county Laseiun Yak milk sample for gradient dilution to 10^-3Adding 2ml of diluent into 20ml of litmus milk culture medium, then putting the litmus milk culture medium into an incubator at 37 ℃ for culture, and taking the litmus milk culture medium out when the culture medium is acidified, solidified and pink.

② culturing single bacterial colony: diluting the litmus milk culture medium bacterial liquid to 10^-5-10^-6Inoculating the mixture to a BL agar culture medium containing 10ppm of cycloheximide and 10ppm of colistin sulfate, and culturing at 37 ℃ for 24-48 h until bacterial colonies are formed.

Screening acid-producing strains: adding 0.2% CaCO into the single colony obtained by primary screening₃BL agar medium in streak cultureCulturing for 24-48 h, and observing whether a transparent ring exists after the culture is finished.

Purifying acid-producing strains: and selecting a single colony with a large transparent circle, and streaking and purifying. The purification was repeated 3 times.

Gram staining preliminary identification: gram staining is carried out on the purified acid-producing strain, and gram positive bacilli with purple thalli are screened for subsequent research.

EXAMPLE 2 preparation of gram-Positive Bacillus fermentation broth

Activating the strain: sucking 1ml of gram-positive bacillus liquid stored in glycerin tube with pipette, inoculating to 9ml of MRS liquid culture medium, activating and culturing at 37 ℃ for 24 h.

② seed liquid culture: 1ml of gram-positive bacillus activation culture solution is sucked by a pipette gun, inoculated into 9ml of MRS liquid culture medium and cultured for 24 hours at 37 ℃.

③ fermentation: the culture medium was inoculated into 45ml of MRS-CHOL (Cholesterol MRS medium) liquid medium by sucking 5ml of gram-positive bacillus seed culture medium with a pipette and cultured at 37 ℃ for 48 hours.

Fourthly, centrifuging the fermentation liquor: the fermentation broth after 48h fermentation was centrifuged at 12000rpm for 10min and the supernatant was collected. Thus obtaining the supernatant of the fermentation liquor of gram-positive bacilli.

Example 3 determination of the Cholesterol-lowering Capacity of gram-Positive bacilli

Drawing of standard curve of cholesterol

Taking 0, 0.1, 0.2, 0.3, 0.4 and 0.5mL of cholesterol storage solution with the concentration of 1mg/mL respectively in a 10mL clean test tube, carrying out water bath at 60 ℃, adding 2mL of ready-prepared o-phthalaldehyde color developing agent after ethanol is completely volatilized, standing at room temperature for 10min after uniform oscillation, slowly adding 1mL of concentrated sulfuric acid, uniformly mixing, standing for 20min, and carrying out color comparison at the wavelength of 560 nm. The abscissa is cholesterol concentration, and the ordinate is absorbance, and a standard curve is drawn. The resulting standard curve is shown in FIG. 1.

The supernatant of the gram-positive bacillus fermentation broth prepared in example 2 was assayed for its ability to reduce cholesterol, and the cholesterol content of the fermentation broth was determined by the o-phthalaldehyde method.

Cholesterol medium without inoculated gram-positive bacilli is used as a blank control group. The cholesterol reduction rate of gram-positive bacilli was then calculated according to the following formula. The results are shown in Table 2.

X＝(1-A/B)*100％；

Wherein: cholesterol reduction rate of X-gram positive bacilli;

a-the cholesterol content in the supernatant of a fermentation broth of gram-positive bacilli;

B-Cholesterol content in supernatant of blank control group.

TABLE 2 reduction of cholesterol by gram-positive bacilli

Strain numbering	Rate of cholesterol reduction	Strain numbering	Rate of cholesterol reduction
				LMLC4560-1	87.3％	LMLC4560-11	41.3％
LMLC4560-2	18.0％	LMLC4560-12	39.0％
				LMLC4560-3	22.7％	LMLC4560-13	40.6％
LMLC4560-4	20.1％	LMLC4560-14	20.6％
				LMLC4560-5	20.8％	LMLC4560-15	35.2％
LMLC4560-6	5.6％	LMLC4560-16	30.5％
				LMLC4560-7	32.3％	LMLC4560-17	23.5％
LMLC4560-8	29.6％	LMLC4560-18	35.2％
				LMLC4560-9	18.0％	LMLC4560-19	48.7％
LMLC4560-10	22.7％	LP-23	36.5％

The results as shown in table 2 show: 19 gram-positive bacilli separated, screened and purified from yak milk and lactobacillus plantarum LP-23 owned by the laboratory, 10 strains with stronger cholesterol-lowering capability in 20 strains in total are respectively: LMLC4560-1, LMLC4560-7, LMLC4560-11, LMLC4560-12, LMLC4560-13, LMLC4560-15, LMLC4560-16, LMLC4560-18, LMLC4560-19 and LP-23.

The LP-23, Lactobacillus plantarum LP-23, was deposited in China Center for Type Culture Collection (CCTCC) at 6/2/2020 with a preservation number of CCTCC NO: M2020175.

The lactobacillus plantarum LP-23 is obtained by separation and purification in the following way:

enriching strains: taking 1ml of milk sample of certain milk factory in Yichang city, Hubei province of China to carry out gradient dilution to 10^-3Adding 2ml of diluent into 20ml of litmus milk culture medium, then putting the culture medium into a 37 ℃ incubator for culture, and taking the culture medium out when the culture medium is acidified, solidified and pink;

② culturing single bacterial colony: diluting the litmus milk culture medium bacterial liquid to 10^-5-10^-6Inoculating the strain to a BL agar culture medium containing 10ppm of cycloheximide and 10ppm of colistin sulfate, and culturing at 37 ℃ for 24-48 h until a bacterial colony is formed;

screening acid-producing strains: adding 0.2% CaCO into the single colony obtained by primary screening₃Performing streak culture in BL agar culture medium for 24-48 h, and observing whether a transparent ring exists after finishing the culture;

purifying acid-producing strains: the single colony with the largest clearing circle, i.e., Lactobacillus plantarum LP-23 strain, was selected.

16SrDNA sequencing and strain identification are carried out on the lactobacillus plantarum LP-23 strain. The primers used were: forward primer 27F: 5'-AGAGTTTGATCATGGCTCAG-3', respectively; reverse primer 1492R: 5'-ACGGCTACCTTGTTACGACTT-3' are provided. The PCR reaction system is as follows: 25 μ Ι mix; 1 μ l of sample; 20 μ l ddH₂O; 2. mu.l of 27F; 2 μ l 1492R; the PCR reaction temperature: at 98 ℃ for 2 min; 98 ℃, 10s, 58 ℃, 10s, 72 ℃, 7s, 35 ×; 72 ℃ for 2 min; storing at 4 ℃. For PCR productsFormulation sequencing, the gene sequence of the 16SrDNA of the lactobacillus plantarum LP-23 strain is shown as SEQ ID NO.1 as follows:

example 4 determination of tolerance of Artificial digestive juice of gram-positive bacilli

The 10 gram-positive bacilli selected in example 3, which had a greater cholesterol-lowering ability, were tested for their resistance to artificial digestive juices.

Preparation of artificial digestive juice

Artificial gastric juice: NaCl 0.20g/100mL, pepsin 0.35g/mL, adjusting pH to 2.5 with 1mol/L HCl, and then filtering and sterilizing for later use.

Artificial intestinal juice: mixing the following solution A and solution B at a ratio of 2:1 to obtain the artificial intestinal juice. A. Pancreatic juice: 1.1g/100mL of sodium bicarbonate, 0.2g/100mL of NaCl and 0.1g/100mL of trypsin, adjusting the pH value to 8.0, and filtering and sterilizing for later use. B. Bile juice: 1.8g/100mL of bile acid salt, adjusting the pH value to 6.8, and filtering and sterilizing for later use.

② measurement of tolerance to artificial digestive juice

Selecting single colony of gram-positive bacillus, culturing in MRS liquid culture medium overnight, inoculating 1mL culture solution into 9mL artificial gastric juice or artificial intestinal juice, culturing at 37 deg.C for 2 hr, and determining viable count. Gram positive bacilli inoculated in 9mL MRS liquid medium served as positive control.

Survival (%). percent (viable count in artificial digestive juice/viable count in control). 100%

TABLE 3 survival rate of gram-positive bacilli treated with artificial gastric juice and artificial intestinal juice

As shown in table 3 above, gram-positive bacillus LMLC4560-1 has higher survival rates in both artificial gastric fluid and artificial intestinal fluid, reaching 96.35% and 92.36%, respectively.

EXAMPLE 5 measurement of DPPH.radical scavenging efficiency of supernatant of gram-positive Bacillus fermentation broth

The supernatant of fermentation broth of 10 gram-positive bacilli selected in example 3, which had a high cholesterol-lowering ability, was assayed for DPPH.radical clearance. The measurement method is as follows:

activating the strain: sucking 1ml of gram-positive bacillus liquid stored in glycerin tube by using a pipette, inoculating into 9ml of MRS liquid culture medium, and performing activated culture at 37 ℃ for 24 h.

② seed liquid culture: 1ml of the activated culture medium was aspirated by a pipette, and the activated culture medium was inoculated into 9ml of MRS liquid medium and cultured at 37 ℃ for 24 hours.

③ fermentation: 5ml of the seed culture solution is sucked by a pipette gun, inoculated into 45ml of MRS liquid culture medium and cultured for 24h at 37 ℃.

Preparing supernatant of fermentation liquor: and centrifuging the fermentation liquor fermented for 24 hours at 5000rpm for 4min, and collecting the supernatant. Thus obtaining the supernatant of the fermentation liquor of gram-positive bacilli.

Fifthly, taking 3mL of fermentation liquor to be measured and mixing with 0.16mmol/L DPPH solution with the same volume (A1 tube); mixing equal volume of absolute ethanol and 0.16mmol/L DPPH solution (A2 tube); taking absolute ethyl alcohol with the same volume and uniformly mixing the absolute ethyl alcohol with a solution to be detected (A3 tube); after 30 ℃ dark reaction for 30min, the absorbance values of tubes A1, A2 and A3 were measured at 519nm using distilled water as a blank and recorded as ODA1, ODA2 and ODA3, respectively. DPPH radical clearance was calculated according to the following formula:

DPPH-clearance (OD)_A2+OD_A3-OD_A1)/OD_A2×100％。

The results are shown in Table 4.

TABLE 4 clearance of DPPH free radical from gram-positive bacilli fermentation broth supernatant

As shown in Table 4 above, the supernatant obtained by fermentation of gram-positive bacillus LMLC4560-1 has a high DPPH free radical scavenging rate of 90.35%.

Example 6 Strain identification

The obtained gram-positive bacterium LMLC4560-1 strain is inoculated on a BL solid medium for streak culture, and the colony morphology is observed. On BL medium, the colony is light yellow, flat and round, neat in edge, opaque, 1.5mm + -0.3 mm. The colony morphology is shown in FIG. 2.

A single colony was picked up and dissolved in 1ml of sterile water, and the cell morphology was observed under a microscope. The identification result shows that: the obtained gram-positive bacterium LMLC4560-1 strain does not move, has no spore, flagella and capsule, and the thallus is in a short rod shape with one end being blunt and round and the other end being smaller and exists in pairs. The morphological characteristics of the strain are shown in figure 3.

16SrDNA sequencing and strain identification are carried out on the LMLC4560-1 strain. The primers used were: forward primer 27F: 5'-AGAGTTTGATCATGGCTCAG-3', respectively; reverse primer 1492R: 5'-ACGGCTACCTTGTTACGACTT-3' are provided. The PCR reaction system is as follows: 25 μ Ι mix; 1 μ l of sample; 20 μ l ddH 2O; 2. mu.l of 27F; 2 μ l 1492R; the PCR reaction temperature: at 98 ℃ for 2 min; 98 ℃, 10s, 58 ℃, 10s, 72 ℃, 7s, 35 ×; 72 ℃ for 2 min; storing at 4 ℃. The PCR product dosage form is sequenced, and the gene sequence of the 16SrDNA of the LMLC4560-1 strain is determined as shown in SEQ ID NO.2 as follows:

the LMLC4560-1 strain is named as Weissella sinus LMLC4560-1 strain (Weissella cibaria LMLC4560-1) and preserved, wherein the Weissella sinus LMLC4560-1 strain (Weissella cibaria LMLC4560-1) is preserved in China Center for Type Culture Collection (CCTCC) at 7/2 of 2021 with the preservation number of CCTCC NO: M2021811.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Sequence listing

<110> Hubei Angel biological group Co., Ltd

<120> lactic acid bacteria with cholesterol-reducing performance and application thereof

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<213> Lactobacillus plantarum (Lactobacillus plantarum)

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ctaatacatg caagtcgaac gaactctggt attgattggt gcttgcatca tgatttacat 60

ttgagtgagt ggcgaactgg tgagtaacac gtgggaaacc tgcccagaag cgggggataa 120

cacctggaaa cagatgctaa taccgcataa caacttggac cgcatggtcc gagcttgaaa 180

gatggcttcg gctatcactt ttggatggtc ccgcggcgta ttagctagat ggtggggtaa 240

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cgaagaagtg catcggaaac tgggaaactt gagtgcagaa gaggacagtg gaactccatg 660

tgtagcggtg aaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctgtctggt 720

ctgtaactga cgctgaggct cgaaagtatg ggtagcaaac aggattagat accctggtag 780

tccataccgt aaacgatgaa tgctaagtgt tggagggttt ccgcccttca gtgctgcagc 840

taacgcatta agcattccgc ctggggagta cggccgcaag gctgaaactc aaaggaattg 900

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tacgatccga aaaccttctt cactcacgcg gcgttgctcg gtcagacttt cgtccattgc 1080

cgaagattcc ctactgctgc ctcccgtagg agtttgggcc gtgtctcagt cccaatgtgg 1140

ccgatcaccc tctcaggtcg gctatgcatc gtagccttgg tgagccgtta cctcaccaac 1200

tagctaatgc accgcgggtc catccatcag cgacacccga aagcgccttt caaatcaaaa 1260

ccatgcggtt tcgattgtta tacggtatta gcacctgttt ccaagtgtta tccccttctg 1320

atgggcaggt tacccacgtg ttactcaccc gttcgccact cttctttttc cggtggagca 1380

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Claims

1. A Weissella cibaria strain is Weissella cibaria LMLC4560-1 strain (Weissella cibaria LMLC4560-1), which is preserved in China Center for Type Culture Collection (CCTCC) at 7, month and 2 days in 2021, and the preservation number is CCTCC NO: M2021811.

2. The Weissella sinus strain of claim 1, wherein the Weissella sinus LMLC4560-1 strain has a 16S rDNA gene sequence as set forth in SEQ ID No. 2.

3. A fermentation preparation method of Weissella cibaria microbial inoculum is characterized by comprising the following steps: culturing the Weissella cibaria strain of claim 1 or 2.

4. The method of claim 3, comprising the steps of:

(1) culturing the Weissella sinus strain of claim 1 or 2 in scale-up;

5. A microbial inoculum obtained by the fermentation production method according to claim 3 or 4.

6. A weissella sinus strain according to claim 1 or 2 or a fermentation product obtained by fermentation of the microbial inoculum according to claim 5.

7. The fermentation product of claim 6, wherein the fermentation product has cholesterol lowering properties;

preferably, the fermentation product has free radical scavenging ability.

8. The fermentation product of claim 6, wherein the fermentation product acts on cholesterol with a cholesterol reduction rate of greater than 87%;

9. The method for producing a fermentation product according to any one of claims 6 to 8, which comprises culturing the Weissella sinus strain according to claim 1 or 2 or the microbial agent according to claim 5.

10. The method of claim 9, comprising the steps of:

(1) amplifying and culturing the Weissella sinus strain of claim 1 or 2 or the microbial inoculum of claim 5;

11. Use of a weissella sinus strain according to claim 1 or 2 or a microbial inoculum according to claim 5 in cholesterol-lowering, lipid-lowering, weight-reducing, antioxidant or anti-ageing products.

12. Use according to claim 11, wherein the product is a food product, a cosmetic product, a nutraceutical product or a pharmaceutical product.