Paracaseicillus casei YYS-K1 and application thereof

Abstract

The application belongs to the technical field of microorganisms, and particularly relates to a paracasei YYS-K1 and application thereof. Lacticaseibacillus paracasei The paracasei YYS-K1 has a preservation number of CGMCC No. 26405 in the China General Microbiological Culture Collection Center, can be applied to preparation of probiotic products, and has the functions of improving intestinal flora structure, relieving alcoholism, and reducing weight.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to Lactobacillus paracasei YYS-K1 and applications thereof. Background Art

[0002] Alcohol, with its unique flavor and taste, has a long history in my country and is a deeply appealing traditional beverage. With the continuous development of human civilization, alcoholic beverage culture has also enriched and developed. Common alcoholic beverages include beer, baijiu, fruit wine, milk wine, and tea wine, which have enormous social and economic benefits. Moderate drinking can improve appetite, aid digestion, relieve fatigue and anxiety, and boost metabolism. However, long-term, excessive consumption is not only harmful but can also cause numerous illnesses, such as alcoholic hepatitis, cirrhosis, and alcoholic fatty liver disease. It can also affect the brain, ultimately leading to varying degrees of brain damage, such as slowed response times and memory loss. Excessive drinking can also strongly stimulate the digestive system, causing ailments such as loss of appetite and malnutrition, or triggering diseases in other organs. In severe cases, it can even paralyze the respiratory muscles and lead to sudden death, posing a significant threat to human health.

[0003] Ethanol, also known as alcohol, is the primary ingredient in various alcoholic beverages. Over 80% of ethanol is metabolized by the liver after entering the body. Alcoholic liver damage, primarily caused by liver cell disruption caused by liver metabolites, is a common disease among alcoholics. With the advancement of science and technology, as well as the growing awareness of health, people are paying more and more attention to the management of excessive alcohol consumption. Therefore, the exploration of the mechanisms of ethanol degradation has become a hot topic in scientific research.

[0004] my country is rich in traditional Chinese medicine (TCM). Researchers have developed a large number of hangover remedies using TCM as research subjects, with some promising results. Most currently available hangover remedies are primarily plant-based. These products can affect the metabolism of ethanol in the human body or inhibit its absorption, thereby sobering up, but they do not actually degrade ethanol. While some products have some efficacy, they may also have toxic side effects that are detrimental to human health. Furthermore, many hangover remedies on the market lack a substantial hangover-relieving effect, merely relieving headaches and exhibiting unsatisfactory results. Therefore, the development of a highly effective, non-toxic, and inexpensive biological hangover remedy is highly desirable.

[0005] Carbohydrates are necessary for the growth of microorganisms. If microorganisms can use ethanol as one of the carbon sources necessary for growth, they can achieve the effect of degrading ethanol. However, there are currently relatively few research reports on this aspect.

[0006] The literature "Screening, identification and optimization of culture conditions of an ethanol-degrading strain [J]. Brewing Technology, 2011 (06): 17-20" screened a strain with relatively high ethanol degradation ability from the sour lees, and was identified as Acetobacter pasteurianus ( Acetobacter pasteurianus ) VII, with an ethanol degradation capacity of 0.72 mL / d.

[0007] The literature "Screening of ethanol-degrading bacteria and evaluation of the hangover-removing efficacy of fermented milk products [J]. Food Science, 2020, 41(02): 107-113" screened out a strain with good ethanol degradation effect from 10 strains of lactic acid bacteria with good fermentation performance, and was identified as Lactobacillus acidophilus ( Lactobacillus acidophilus ), after 24 h of culture, the ethanol degradation rate was about 50%.

[0008] The paper "Screening and Identification of Anaerobic Microorganisms for Rapid Alcohol Relief and Study on Their Ability to Relieve Alcohol [D]. Zhejiang University, 2016" screened a strain of bacteria from Korean kimchi. After culturing at 20% ethanol concentration for 1 hour, the strain's ethanol degradation rate was 21.1%. Sequencing identified the strain as Enterococcus hirae ( Enterococcus hirae ).

[0009] The Chinese invention patent with application number 201911202646.7 discloses a hangover-relieving and liver-protecting composition and product, which includes plant extracts and probiotics. The probiotics include Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium lactis Bi-07, Bifidobacterium lactis Bl-04 and Bifidobacterium lactis HN019. However, it is a combination product. There is a large amount of literature proving that the plant ingredients therein have the effect of sobering up, but it cannot be proved that the probiotics therein have the effect of sobering up, and it does not indicate that any of the strains used can have a good sobering effect when used alone.

[0010] Obesity is one of the major factors affecting human health in today's society. Most of it is caused by excessive food intake, which leads to fat accumulation and weight gain. It is common in the population. Therefore, the development of weight loss products is also a hot topic of current research. Most existing weight loss drugs act on the nerve center and have strong side effects on the human body. Another type of weight loss product - lipase inhibitors - can act without passing through the human nervous system or blood vessels. They do not pose a major safety hazard to human health and are a safe new type of weight loss preparation. Pancreatic lipase is secreted by the pancreas and is a key enzyme in the fat digestion process. Pancreatic lipase inhibitors can inhibit the decomposition of fat by pancreatic lipase in the intestines, reduce the absorption of fatty acids by the small intestinal mucosa, and thus achieve a weight loss effect. Microorganisms are an important source of a variety of enzyme preparations. Microbial metabolites have been widely used in the food industry and have high application value.

[0011] The literature "Screening of Lipase Inhibitor-Producing Bacteria [J]. Chinese Journal of Antibiotics, 2002 (11): 641-643" isolated and screened 8 strains with lipase inhibitory activity from soil samples. Among them, the strain LF with the highest inhibition rate was as high as 80.6%. It was identified as Bacillus and was not an edible probiotic. Summary of the Invention

[0012] In order to overcome the defects of the prior art, the present invention provides a Lactobacillus paracasei and its application. The Lactobacillus paracasei of the present invention can efficiently degrade ethanol and has a high inhibitory effect on pancreatic lipase. It can be used in the development of alcohol detoxification or weight loss products and has important application value.

[0013] In order to solve the above technical problems, the technical solution adopted by the present invention is: Lactobacillus paracasei YYS-K1, classified and named: Lactobacillus paracasei YYS-K1, Latin name: Lacticaseibacillus paracasei , deposited in the General Microbiology Center of China Culture Collection Administration, with the deposit number: CGMCC No. 26405. Deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit date: January 6, 2023.

[0014] The present invention also provides the use of the Lactobacillus paracasei YYS-K1 in preparing a probiotic product.

[0015] The present invention also provides the use of the Lactobacillus paracasei YYS-K1 in preparing acid-resistant and / or bile-resistant food.

[0016] The present invention also provides use of the Lactobacillus paracasei YYS-K1 in preparing an alcohol sobering agent.

[0017] The present invention also provides use of the Lactobacillus paracasei YYS-K1 in preparing weight-loss products.

[0018] The present invention also provides a bacterial agent containing the Lactobacillus paracasei YYS-K1.

[0019] The application has the beneficial effects that the application provides a paracasei YYS-K1 with high ethanol degradation capacity and pancreatic lipase inhibition rate, the ethanol degradation rate of which is 43.9% in an MRS culture medium added with 20% ethanol, and the ethanol degradation rate of which is 48.6% in an MRS culture medium added with 40% ethanol; the fermentation liquor of the paracasei YYS-K1 has a pancreatic lipase inhibition rate of 64.9%, and meanwhile, the paracasei YYS-K1 has good acid resistance and bile salt resistance, and the high ethanol degradation capacity, pancreatic lipase inhibition capacity and acid resistance and bile salt resistance of the paracasei YYS-K1 enable the paracasei YYS-K1 to be applied to the preparation of probiotic products for improving intestinal flora structure, and alcoholism relieving or weight loss products, and the problem that plant raw materials are prone to causing toxic side effects due to complex components is eliminated, and a new direction and more possibilities are provided for the development of alcoholism relieving or weight loss products. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 A calcium-dissolving ring in the separation process of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0021] Figure 2 A colony morphology of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0022] Figure 3 A viable cell count of the paracasei YYS-K1 in the specific embodiment of the application under different ethanol concentrations is shown;

[0023] Figure 4 An ethanol standard curve in the ethanol degradation test of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0024] Figure 5 A 4-nitrophenol (PNP) standard curve in the pancreatic lipase inhibition test of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0025] Figure 6 A gram staining result of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0026] Figure 7 A 16S rRNA amplification product agarose gel electrophoresis map of the paracasei YYS-K1 in the specific embodiment of the application is shown;

[0027] Figure 8 A phylogenetic tree of the paracasei YYS-K1 in the specific embodiment of the application based on 16S rRNA sequences is shown. DETAILED DESCRIPTION

[0028] To illustrate the technical content, achieved objectives and effects of the present invention in detail, the following description is given in conjunction with the embodiments and accompanying drawings.

[0029] The present invention provides a kind of Lactobacillus paracasei ( Lacticaseibacillus paracasei ) YYS-K1, deposited in the General Microbiology Center of China Culture Collection Administration, deposit number: CGMCC No. 26405, deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit date: January 6, 2023.

[0030] Example 1: Isolation of Lactobacillus paracasei YYS-K1

[0031] Aseptic sampling was performed from homemade bayberry wine at the farmer's home. 5 g of bayberry wine sample was placed in a sterile homogenizing bag using the spread plate method. 45 mL of 0.85% saline was added and the sample was thoroughly patted and mixed to obtain the sample. 100 μL of the sample was then drawn for a 10-fold series of gradient dilutions. 100 μL of the sample was drawn for 10-fold series of gradient dilutions. -4 , 10 -5 , 10 -6 100 μL of the sample was spread on an MRS plate containing 2.5% CaCO3 and incubated at 37℃ for 24 h. The colonies with good growth and large calcium dissolution zone were selected (see Figure 1 ), and repeatedly isolated and purified by plate streak separation method until a single colony was obtained (see Figure 2 The isolated strain was named YYS-K1, numbered, and stored in a -80℃ bacteria bank after adding glycerol.

[0032] Example 2: Ethanol degradation test of Lactobacillus paracasei YYS-K1

[0033] 2.1 Analysis of ethanol tolerance

[0034] The Lactobacillus paracasei YYS-K1 strain was activated to the second generation and inoculated into MRS medium supplemented with 0%, 10%, 20%, 30%, 40%, and 50% anhydrous ethanol at a 3% inoculum size. Three replicates were used in each group and cultured at 37°C for 24 h. After the incubation period, the number of viable bacteria was determined by the plate pouring method and the ethanol tolerance was analyzed. The results are shown in Figure 3 shown.

[0035] 2.2 Solution preparation

[0036] Ethanol standard solution: Accurately pipette anhydrous ethanol and add it to distilled water to prepare ethanol standard solutions with volume fractions of 0%, 10%, 30%, 50%, 70%, and 90%, respectively.

[0037] Potassium dichromate-concentrated sulfuric acid solution: Weigh 1 g of potassium dichromate and dissolve it in 25 mL of distilled water. Slowly add 4 mL of concentrated sulfuric acid (95%~98%) while stirring.

[0038] 2.3 Drawing of ethanol standard curve

[0039] Add 1.0 mL of 0%, 10%, 30%, 50%, 70%, and 90% ethanol standard solutions to each test tube. Add 3.0 mL of potassium dichromate-concentrated sulfuric acid solution to each test tube, shake thoroughly, and secure the lid. Place the test tube in a boiling water bath for 10 minutes to fully oxidize the potassium dichromate-concentrated sulfuric acid solution. Remove the test tube and measure the absorbance of the potassium dichromate-concentrated sulfuric acid solution at a wavelength of 610 nm. Plot a standard curve with ethanol concentration as the horizontal axis and absorbance as the vertical axis (see Figure 4 ).

[0040] 2.4 Detection and analysis of test samples

[0041] Prepare ethanol-MRS liquid medium with 20% and 40% ethanol by volume, respectively. Transfer 5 mL of liquid medium to a test tube and add 200 μL of second-generation activated YYS-K1 bacterial culture. Mix thoroughly and incubate on a shaker at 37°C for 24 hours. Ethanol-MRS liquid medium without bacterial culture was used as a control. Residual ethanol was determined using the potassium dichromate-concentrated sulfuric acid method to analyze the ethanol degradation ability of the strain. Each sample was replicated in triplicate, and absorbance was measured at 610 nm.

[0042] The linear relationship between ethanol concentration and OD610 nm absorbance was obtained from the measured standard curve. Ethanol degradation rate = (original ethanol concentration - residual ethanol concentration) / original ethanol concentration × 100%. Sample testing showed that the ethanol degradation rate of YYS-K1 could reach 43.9% when the ethanol concentration was 20%, and the ethanol degradation rate could reach 48.6% when the ethanol concentration was 40%.

[0043] MRS liquid medium: 20.0 g glucose, 10.0 g tryptone, 10.0 g beef extract, 5.0 g yeast extract powder, 1.0 mL Tween 80, 2.0 g dipotassium hydrogen phosphate, 2.0 g ammonium citrate, 5.0 g anhydrous sodium acetate, 0.5 g magnesium sulfate, 0.25 g manganese sulfate monohydrate, 1 L deionized water, pH 6.5 (add 1.5% agar for MRS solid medium).

[0044] The Lactobacillus paracasei YYS-K1 provided by the present invention has good ethanol degradation ability and can be used for preparing an alcohol sobering agent.

[0045] Example 3: Lactobacillus paracasei YYS-K1 pancreatic lipase inhibition test

[0046] 3.1 Solution preparation

[0047] Tris-HCl buffer: Accurately weigh 6.05 g of Tris and dissolve it in 1 L of distilled water. Then add 0.1% gum arabic powder and 0.2% sodium deoxycholate and adjust the pH to 8.0.

[0048] Pancreatic lipase solution (PPL): Weigh 240 mg of pancreatic lipase and dissolve it in 200 mL of Tris-HCl buffer. Mix well and allow to stand. Centrifuge at 6000 rpm for 5 min. Collect the supernatant and store at -20°C.

[0049] Substrate solution: Weigh 20 mg of 4-nitrophenyl palmitate (PNPP) and dissolve it in 5 mL of isopropanol. Then dilute to 100 mL with Tris-HCl buffer.

[0050] 3.2 Drawing of PNP standard curve

[0051] Accurately weigh 7.0 mg of 4-nitrophenol (PNP) standard, dissolve it in dimethyl sulfoxide (DMSO), and dilute to 10 mL to prepare a 5 mmol / L stock solution. Dilute the stock solution PNP to 0.05 mmol / L, 0.25 mmol / L, 0.1 mmol / L, 0.2 mmol / L, and 0.3 mmol / L using Tris-HCl buffer. Measure the absorbance of different concentrations of PNP at 405 nm. Plot a standard curve (see ) with PNP concentration as the horizontal axis and absorbance as the vertical axis. Figure 5 ).

[0052] 3.3 Sample detection and analysis

[0053] Lactobacillus paracasei YYS-K1 was activated to the second generation and centrifuged at 8000 rpm for 10 minutes. The supernatant was the test sample solution. The experiment was divided into four groups, A, B, C, and D. The dosages of the reactants are shown in Table 1. Standard 96-well plates were used, with five replicates per group. The experimental conditions were designed to simulate the human body environment. Tris-HCl buffer (pH 8.0), the test sample solution, and pancreatic lipase solution were added sequentially, mixed thoroughly, and incubated in a 37°C water bath for 10 minutes. After the reaction, the 96-well plate was removed and the substrate solution (PNPP) was added. The mixture was thoroughly mixed and the reaction was incubated in a 37°C water bath for 20 minutes. PNPP can be hydrolyzed by pancreatic lipase to produce PNP, which has a maximum absorbance at 405 nm. The absorbance was measured and the inhibition rate of Lactobacillus paracasei YYS-K1 against pancreatic lipase was calculated using the following formula:

[0054] Inhibition rate = [1-(CD / AB)] × 100%

[0055] Where A is the absorbance value of the control group, B is the absorbance value of the control blank group, C is the absorbance value of the sample group, and D is the absorbance value of the sample blank group.

[0056] The measured standard curve shows a linear relationship between PNP and OD405 nm absorbance. Sample testing shows that the inhibition rate of YYS-K1 on pancreatic lipase PPL can reach 64.9%.

[0057] The Lactobacillus paracasei YYS-K1 provided by the present invention has a good pancreatic lipase inhibitory effect and can be applied to the development of weight loss products.

[0058] Table 1

[0059]

[0060] Example 4: Identification of Lactobacillus paracasei YYS-K1

[0061] 4.1 Physiological and biochemical tests

[0062] The screened and purified strain YYS-K1 was subjected to Gram staining (see Figure 6 ) and catalase tests, and physiological and biochemical parameters were measured. The results were compared with the "Bergey's Manual of Systematic Bacteriology, Eighth Edition" for preliminary identification of the bacterial species. The tests revealed that the screened strain, YYS-K1, was Gram-positive with a purple stain. Its cells were rod-shaped, catalase and oxidase tests were negative, and it did not form spores.

[0063] 4.2 16S rRNA identification

[0064] The DNA of the unknown strain YYS-K1 was extracted according to the instructions of the bacterial gene DNA extraction kit, and the 16S rRNA gene was amplified by PCR using the DNA as a template.

[0065] Amplification primers use universal primers:

[0066] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO. 1).

[0067] 1492R: 5'-TACGGCTACCTTGTTACGACTT-3' (SEQ ID NO. 2).

[0068] PCR reaction system: DNA 2 μL, 27F 2 μL, 1492R 2 μL, Premix Ex Taq 25 μL, ddH2O 19 μL.

[0069] PCR reaction conditions were as follows: pre-denaturation at 94°C for 3 min; 30 cycles of denaturation at 94°C for 30 s, annealing at 55°C for 30 s, and extension at 72°C for 1 min; and a final extension at 72°C for 5 min.

[0070] After YYS-K1 extraction of DNA genome, 16S rRNA amplified fragments are shown in Figure 7 The PCR amplification products were then sent for DNA sequencing (Guangzhou Qingke Biotechnology Co., Ltd.). The sequencing results were searched for similar sequences in the NCBI database using Blast software. The obtained sequences were aligned with the 16S rRNA gene sequences of related species obtained from the gene bank, and a phylogenetic tree was constructed using Mega7.0 software. The results are shown in Figure 8 shown.

[0071] The results of 16S rRNA gene sequencing are as follows:

[0072]

[0073] Based on comprehensive analysis of the cell morphology, physiological and biochemical characteristics, 16S rRNA gene sequence and other data of the strain, and with reference to the Bergey's Manual of Systematic Bacteriology, 8th Edition, the strain was identified as Lactobacillus paracasei ( Lacticaseibacillus paracasei ).

[0074] Example 5: Acid resistance and bile salt resistance test of Lactobacillus paracasei YYS-K1

[0075] 5.1 Simulated gastric juice acid resistance test

[0076] Weigh 0.2 g of NaCl and 0.35 g of pepsin and dissolve them in an appropriate amount of distilled water. Three aliquots were prepared. The pH values ​​were adjusted to 1.5, 2.5, and 3.5, respectively, with 1.0 mol / L hydrochloric acid, and the volume was brought to the mark in a 100 mL volumetric flask. A 2% inoculum of the fermentation broth of Lactobacillus paracasei YYS-K1, cultured for 24 h after activation, was inoculated into simulated gastric acid solutions of varying pH values. After incubation in a 37°C waterbath for 0, 2, and 4 h, the broth was serially diluted 10-fold, shaken, and 100 μL of the culture was poured onto plates. The culture was then incubated at 37°C for 24 h, and the viable YYS-K1 cells were counted. The results are shown in Table 2.

[0077] Table 2

[0078]

[0079] Table 2 shows that the viable counts of Lactobacillus paracasei YYS-K1 varied in simulated gastric fluid at different pH values. With increasing pH, the viable count and survival rate increased. In simulated gastric fluid at the same pH, the viable count decreased with increasing treatment time. However, the viable count remained high after 2 hours of treatment in simulated gastric fluid at different pH values, indicating that YYS-K1 can maintain a high survival rate after digestion in human gastric fluid and exhibits excellent tolerance to gastric acid.

[0080] 5.2 Simulated intestinal fluid bile salt tolerance test

[0081] The fermentation broth of Lactobacillus paracasei YYS-K1 activated and cultured for 24 h was inoculated at a 2% inoculum volume into pig bile salt solutions with mass concentrations of 0.03 g / mL, 0.3 g / mL, and 0.5 g / mL, respectively. After being treated in a 37°C water bath for 0, 2, and 4 h, the solution was diluted 10-fold, shaken evenly, and 100 μL of the bacterial solution was poured onto a plate. After incubation at 37°C for 24 h, the number of viable YYS-K1 cells was calculated. The results are shown in Table 3.

[0082] Table 3

[0083]

[0084] As can be seen from Table 3, the viable cell number of Paracaseicillus casei subsp. paracasei YYS-K1 is different at different initial mass concentrations of bile salts in the simulated artificial intestinal fluid, and the survival rate is also different. With the increase of the mass concentration of bile salts in the simulated artificial intestinal fluid, the initial viable cell number gradually decreases, and the survival rate also gradually decreases with the longer treatment time. However, after 2 h of treatment at the bile salt concentration closest to the human body (the bile salt concentration in the human small intestine is generally 0.03-0.3 g / mL), there are still a relatively high number of surviving cells, indicating that YYS-K1 can still have a relatively high number of viable cells after gastrointestinal digestion in the human body to metabolize ethanol.

[0085] The Paracaseicillus casei subsp. paracasei YYS-K1 has good gastric acid resistance and bile salt resistance, can improve the intestinal flora structure, and is used for preparing acid-resistant and / or bile salt-resistant food, and has high commercial application value.

[0086] Example 6:

[0087] A microbial agent (probiotic product) comprises Paracaseicillus casei subsp. paracasei YYS-K1 and a protective agent; the protective agent is composed of 100 g / L skimmed milk powder, 30 mL / L glycerol, 150 g / L trehalose and 10 g / L L-glutamic acid sodium.

[0088] The preparation method of the above microbial agent is as follows:

[0089] S1, inoculate Paracaseicillus casei subsp. paracasei YYS-K1 into a culture medium sterilized at 121℃ for 15 min at an inoculation amount of 3% of the mass of the culture medium, then culture at 37℃ for 20 h, centrifuge at 6000 r / min for 20 min at 4℃, discard the supernatant, wash with pH=7.2 phosphate buffer for 2-4 times to obtain bacterial slurry, resuspend with a protective agent to obtain a suspension with a bacterial liquid concentration of 10 10 CFU / mL;

[0090] The culture medium is composed of 0.5% glucose, 1.5% tryptone, 0.3% yeast extract and the balance of water in terms of the total mass of the culture medium, and the pH is 6.8;

[0091] S2, pre-culture the suspension of S1 at 37℃ for 60 min, and then freeze-dry to obtain a microbial agent containing active Paracaseicillus casei subsp. paracasei YYS-K1 with a content of more than 1×10 10 CFU / g.

[0092] Example 7:

[0093] A probiotic product comprises Paracaseicillus casei subsp. paracasei YYS-K1 and other ingredients. In the probiotic product, the viable cell number of Paracaseicillus casei subsp. paracasei YYS-K1 is not less than 1×106 CFU / mL or 1 x 10 6 CFU / g. The ingredients include one or several of prebiotics, fillers, acidifiers, solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavorings, preservatives, coating materials, fragrances, anti-adhesion agents, integration agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, antifoaming agents, thickening agents, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculants, and filter aids.

[0094] The dosage form of the probiotic product can be a solid beverage, a liquid beverage, a pressed candy, a granule, a capsule, a tablet, a pill, or an oral liquid.

[0095] In this embodiment, a probiotic product (Paracasei YYS-K1 solid beverage) with a dosage form of a solid beverage is provided, which comprises, by weight, 10 parts of active bacteria powder, 24.8 parts of xylitol, 24 parts of whole milk powder, 12 parts of fermented mulberry powder, 23 parts of malt dextrin, 5.6 parts of galacto-oligosaccharide, 0.1 part of fructo-oligosaccharide, and 0.5 part of anhydrous citric acid.

[0096] The preparation method of the solid beverage is as follows:

[0097] S1, the Paracasei YYS-K1 strain is cultured with a liquid culture solution, the bacterial cells are collected and washed, and the auxiliary materials (the protective agent of Example 6) are added to prepare the active bacteria powder by drying;

[0098] S2, the active bacteria powder, xylitol, whole milk powder, fermented mulberry powder, malt dextrin, galacto-oligosaccharide, fructo-oligosaccharide, and anhydrous citric acid are uniformly mixed to obtain a Paracasei YYS-K1 solid beverage with a viable bacterial count of not less than 1 x 10 8 CFU / g.

[0099] The experimental operation methods not explicitly indicated in all the embodiments are conventional experimental operation methods in the art, and the reagents or instruments involved can be commercially available from regular channels.

[0100] In summary, there is no report on the functional strain of Paracaseicillus casei for alcohol elimination or pancreatic lipase inhibition, and the Paracaseicillus casei YYS-K1 provided by the present application is different from the species of Bacillus pasteurii and Lactobacillus acidophilus, and is different from the species of Enterococcus hirae, which is different from the existing probiotic alcohol elimination preparation and needs to be used in combination, and can achieve good ethanol degradation capacity when used alone. The ethanol degradation rate is 43.9% when 20% ethanol is added in the MRS medium, and the ethanol degradation rate is 48.6% when 40% ethanol is added in the MRS medium. Meanwhile, the Paracaseicillus casei YYS-K1 has high pancreatic lipase inhibition activity and good acid and bile salt resistance, and can be applied to the preparation of probiotic products for improving intestinal flora structure, and can play a more long-term and stable role when applied to alcohol elimination products or weight loss products. Meanwhile, as a single component, the Paracaseicillus casei YYS-K1 does not have the problem of toxic side effects caused by the complex components of plant raw materials or the unknown interaction between components after compounding, and provides a new direction and more possibilities for the development of alcohol elimination or weight loss products.

[0101] The above description is only an embodiment of the present application, and does not limit the patent scope of the present application, and any equivalent transformation or direct or indirect application in the related technical field based on the content of the specification and drawings of the present application is also included in the patent protection scope of the present application.

Claims

1. Lactobacillus paracasei YYS-K1, characterized in that Classification name: Lactobacillus paracasei, Latin name: Lacticaseibacillus paracasei , deposited in the General Microbiology Center of China Culture Collection Administration of Microorganisms, with the deposit number: CGMCC No. 26405.

2. Application of Lactobacillus paracasei YYS-K1 according to claim 1 in preparing a probiotic product.

3. Application of Lactobacillus paracasei YYS-K1 according to claim 1 in preparing acid-resistant and / or bile-resistant food.

4. Use of the Lactobacillus paracasei YYS-K1 according to claim 1 in preparing an alcohol sobering agent.

5. Application of Lactobacillus paracasei YYS-K1 according to claim 1 in preparing weight-loss products.

6. A bacterial agent, characterized in that The invention comprises the Lactobacillus paracasei YYS-K1 according to claim 1.

7. The microbial agent according to claim 6, characterized in that The viable count of the Lactobacillus paracasei YYS-K1 is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

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