Lactobacillus paracasei and uses thereof

Abstract

The present application relates to the field of microorganism, discloses a lactobacillus paracasei and application thereof.The strain can degrade cholesterols and inhibit the growth and reproduction of streptococcus mutans; in addition, the strain not only has strong acid production capacity, but also can ferment butter under low temperature conditions, and the butter obtained has rich and harmonious flavor.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microorganisms, in particular to a lactobacillus paracasei and application thereof. BACKGROUND

[0002] Hypercholesterolemia seriously threatens the life and health of human beings, and its drug treatment not only has unstable drug efficacy, but also can cause a series of side effects, so it is urgent to find a new method for reducing serum cholesterol. Studies have shown that lactic acid bacteria and fermented milk products have certain ability to regulate blood lipids and reduce serum cholesterol.

[0003] The oral cavity is the starting part of the digestive tract, an important immune organ of the human body, and the first barrier to defend the human body. However, Streptococcus mutans exists in the oral cavity, which not only plays a very important role in the pathogenesis of dental caries, but also can cause gastrointestinal inflammation once it enters the digestive tract. Therefore, it has very important practical significance to inhibit Streptococcus mutans while decomposing cholesterol.

[0004] Although lactobacillus paracasei is often used to prepare drugs for treating or preventing oral diseases caused by high cholesterol or Streptococcus mutans, the existing available strains are few, the ability to lower blood lipids or inhibit pathogenic bacteria is limited, and most importantly, the existing lactobacillus paracasei cannot have both the functions of decomposing cholesterol and inhibiting Streptococcus mutans.

[0005] Therefore, it is urgent to develop a lactobacillus paracasei which can simultaneously decompose cholesterol and inhibit Streptococcus mutans. SUMMARY

[0006] The present application aims to overcome the above-mentioned problems existing in the prior art, and provides a lactobacillus paracasei and application thereof.

[0007] In order to achieve the above-mentioned purpose, the present application provides a lactobacillus paracasei in a first aspect Lactobacillus paracasei The preservation number of the lactobacillus paracasei is CGMCC No.33513.

[0008] The present application provides a microbial agent containing the lactobacillus paracasei as described above in a second aspect.

[0009] The present application provides application of the lactobacillus paracasei as described above in preparing a preparation for promoting cholesterol decomposition in a third aspect.

[0010] The present application provides application of the lactobacillus paracasei as described above in preparing a preparation for inhibiting the growth and reproduction of Streptococcus mutans in a fourth aspect.

[0011] The present application provides application of the lactobacillus paracasei as described above in low-temperature fermentation to prepare butter in a fifth aspect.

[0012] The application provides fermented butter obtained by the application.

[0013] By the technical scheme, the application has at least the following beneficial effects:

[0014] (1) The Lactobacillus paracasei provided by the application can degrade cholesterol and inhibit the growth and reproduction of Streptococcus mutans.

[0015] (2) The Lactobacillus paracasei provided by the application has good acid production performance, and the product obtained by fermentation of the strain has a lower pH value under the same conditions, and the fermented butter product has a more mellow and coordinated flavor.

[0016] Biological preservation

[0017] The strain provided by the application is named Lactobacillus paracasei Lactobacillus paracasei , and was preserved in the China General Microbiological Culture Collection Center (CGMCC) on February 11, 2025, with a preservation number of CGMCC No. 33513 and a preservation address of No. 3, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is a colony morphology diagram of the Lactobacillus paracasei CCNH411 provided by the application;

[0019] Figure 2 is a gram staining diagram of the Lactobacillus paracasei CCNH411 provided by the application. DETAILED DESCRIPTION

[0020] The endpoints of the ranges and any values disclosed herein are not limited to the precise values recited as the exact dimensions are not considered critical for the application. The endpoints of the ranges and any values are only approximations as the ranges and values are understood to include values approximating the value in and around the ranges. The endpoints of the ranges of values and the values thereof, both inclusive and exclusive, can be combined with one or more other endpoints to form new ranges or values not expressly mentioned herein. These new ranges and values are also contemplated as being within the scope of the application.

[0021] The inventors of the application accidentally isolated a Lactobacillus paracasei strain from fermented milk in Xinjiang and named it strain CCNH411. It was found that the strain can decompose cholesterol, has an inhibitory effect on Streptococcus mutans, and has strong acid production capacity. The fermentation process can be carried out at low temperature, and fermented butter with rich flavor and outstanding flavor characteristics can be obtained.

[0022] Based on the above findings, the first aspect of the application provides a Lactobacillus paracaseiLactobacillus paracasei The Lactobacillus paracasei has a preservation number of CGMCC No. 33513.

[0023] In the present application, "CCNH411" is the number of Lactobacillus paracasei given by the inventor during the research process, and "CGMCC No. 33513" is the preservation number of the strain, both of which represent the same strain, and both can be used interchangeably hereinafter.

[0024] In the present application, the 16S rDNA sequence of the Lactobacillus paracasei CCNH411 is shown in SEQ ID NO. 1.

[0025] SEQ ID NO. 1:

[0026] TCGAACGAGTTCTCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGA

[0027] The second aspect of the present application provides a bacterial agent containing the Lactobacillus paracasei as described above.

[0028] The specific type of the fermenting agent is not particularly limited in the present application, and any type of fermenting agent commonly used in the art is suitable for use in the present application. For example, it can be a liquid fermenting agent, a solid fermenting agent, or a semi-solid fermenting agent, etc.

[0029] In some preferred embodiments of the present application, the fermenting agent can be a liquid fermenting agent. For example, Lactobacillus paracasei can be cultured in a liquid medium, and the obtained culture solution can be used as a liquid fermenting agent.

[0030] In the present application, the bacterial agent also contains an auxiliary material, which can be an auxiliary material commonly used in the art for the preparation of bacterial agents or fermenting agents, such as a buffer (such as a buffer solution, a culture medium, etc.), a protective agent (such as a freeze-drying protective agent, e.g., skimmed milk powder, trehalose, sodium glutamate, glycerol, etc.), etc., and is preferably glycerol and / or corn oil.

[0031] In the present application, the number of viable Lactobacillus paracasei bacteria is 10 4 -10 12 CFU per 1 mL of auxiliary material.

[0032] Preferably, the content of Lactobacillus paracasei in the fermenting agent provided by the present application can be 10 4 -10 12 CFU / g (liquid fermenting agent can be converted to 1 g = 1 mL).10 4 -10 12 CFU / g means that the content of Lactobacillus paracasei in the fermenting agent is at the order of magnitude of 10 4 -10 12 CFU / g, for example, at the order of magnitude of 10 4 CFU / g represents a range of greater than or equal to 1×10 4 CFU / g to less than 1×10 5 CFU / g, i.e., such as 1×10 4 CFU / g, 5×10 4 CFU / g, 9.9×10 4 CFU / g, etc. all belong to the order of magnitude of 10 4 CFU / g. Therefore, the content of Lactobacillus paracasei in the method provided by the present application is in the range of greater than or equal to 1×10 4 CFU / g to less than 1×10 13 CFU / g. For example, the content of Lactobacillus paracasei in the fermenting agent provided by the present application can be 1×10 4 CFU / g, 5×10 4 CFU / g, 1×10 5 CFU / g, 5×10 5 CFU / g, 1×10 6CFU / g, 5x10 6 CFU / g, 1x10 7 CFU / g, 5x10 7 CFU / g, 1x10 8 CFU / g, 5x10 8 CFU / g, 1x10 9 CFU / g, 5x10 9 CFU / g, 1x10 10 CFU / g, 5x10 10 CFU / g, 1x10 11 CFU / g, 5x10 11 CFU / g, 1x10 12 CFU / g, 5x10 12 CFU / g, or also can be any two of the above values in the range of any range or value.

[0033] The third aspect of the present application provides the use of Lactobacillus paracasei as described above in the preparation of a preparation for promoting the decomposition of cholesterol.

[0034] Preferably, the cholesterol is glycocholic acid and / or taurocholic acid.

[0035] The fourth aspect of the present application provides the use of Lactobacillus paracasei as described above in the preparation of a preparation for inhibiting the growth and reproduction of Streptococcus mutans.

[0036] Preferably, the Streptococcus mutans is numbered CGMCC 1.2499.

[0037] The fifth aspect of the present application provides the use of Lactobacillus paracasei as described above in the preparation of butter by low-temperature fermentation.

[0038] In the present application, the amount of Lactobacillus paracasei or bacterial agent is such that the number of viable Lactobacillus paracasei in the fermentation raw material is not less than 10 6 CFU / mL.

[0039] Preferably, the fermentation conditions include a temperature of 10-35℃ and a time of 20-100h.

[0040] According to the present application, since the strain provided by the present application has the characteristics of being able to produce acid and ferment butter at a lower temperature, in order to obtain butter with a more concentrated flavor, the temperature of the fermentation is preferably 10-20℃ and the time is preferably 20-50h.

[0041] The sixth aspect of the present application provides the fermented butter obtained by the use as described above.

[0042] The present application further provides the use of Lactobacillus paracasei as described above in dairy products or for improving the flavor of fermented butter.

[0043] The application further provides a composite microbial preparation, which comprises: the Lactobacillus paracasei with the preservation number of CGMCC No. 33513 and the Lactobacillus paracasei with the preservation number of CGMCC No. 33154 (the publication number is CN119752739A) provided by the application.

[0044] The Lactobacillus paracasei with the preservation number of CGMCC No. 33513 provided by the application can ferment butter with richer flavor and better quality under low-temperature conditions, and the Lactobacillus paracasei with the preservation number of CGMCC No. 33154 can improve the types and contents of flavor substances in fermented butter when preparing the fermented butter, so that the fermented butter has outstanding butter aroma, and the lactic acid smell and fruit aroma are coordinated, and the fermentation is carried out under low-temperature conditions (10-20℃), so that the types and contents of flavor substances in the fermented butter are improved, and therefore the Lactobacillus paracasei with the preservation numbers of CGMCC No. 33513 and CGMCC No. 33154 are particularly suitable for low-temperature fermented butter. When preparing butter by fermentation, the order of adding the two Lactobacillus paracasei to raw materials is not limited, and the two Lactobacillus paracasei can be added simultaneously or sequentially, and in order to obtain butter with richer flavor, the two Lactobacillus paracasei are preferably added sequentially.

[0045] The application will be described in detail through examples below. Unless otherwise specified, all reagents or materials used below can be commercially available through regular means.

[0046] Reference strain: Lactobacillus paracasei Lactobacillus paracasei with the number of CICC 25261, purchased from the China Industrial Microbial Culture Collection Center; Lactobacillus rhamnosus Lactobacillus rhamnosus, with the number of ATCC 53103, referred to as LGG, purchased from the American Type Culture Collection;

[0047] Streptococcus mutans Streptococcus mutans with the number of CGMCC 1.2499, purchased from the China General Microbiological Culture Collection Center.

[0048] Essential Morning Crème Fraiche: fat content 36wt%, protein content 1.5wt%, carbohydrate content 2.2wt%.

[0049] MRS liquid medium and MRS solid medium are both purchased from Qingdao Haibo Biology, and the item numbers are HB0384-1 and HB0384-5 respectively.

[0050] Example 1

[0051] The sample was derived from fermented milk in Xinjiang, and lactic acid bacteria were isolated by dilution coating method: 20 g of sample was weighed and added into 180 mL of physiological saline solution and shaken well, and the shaken liquid was sequentially diluted in sterile distilled water at 10 -1 , 10 -2 , 10 -3 , 10 -4 and 10 -5 five gradient dilutions, and then sequentially inoculated on MRS solid medium plate by plate streaking method. After the medium was incubated at 37°C under constant temperature and sealed condition for 2 days, the growth of lactic acid bacteria was observed, different lactic acid bacteria colonies were selected according to luster, transparency and size, and continuous isolation and purification was carried out twice, and strains with lactic acid bacteria phenotype characteristics were isolated.

[0052] The strain was streaked on the plate of MRS solid medium and activated, and incubated at 37°C in an incubator for 48 h. A smooth single colony was picked from the first generation plate and inoculated into a flask containing 30 mL of MRS liquid medium, and after 24 h of incubation at 37°C, a second generation seed liquid was obtained, and the viable cell count reached 10 8 -10 9 CFU / mL. The strain was added into each well of a 96-well deep well plate, one column was inoculated every other column, 1 mL of MRS liquid medium and 20 μL of bacterial liquid were added into each well, and each repeat was three plates. The bacterial body was collected by centrifugation at 3500 rpm for 8 min using a 96-well plate centrifuge, resuspended and washed with sterile water, repeated twice, and then resuspended with 550 μL of sterile water, mixed and used.

[0053] The strain was streaked on the plate of MRS solid medium and activated, and incubated at 37°C in an incubator for 48 h. A smooth single colony was picked from the first generation plate and inoculated into a flask containing 30 mL of MRS liquid medium, and after 24 h of incubation at 37°C, a second generation seed liquid was obtained, and the viable cell count reached 10 8 -10 9 CFU / mL. The strain was added into each well of a 96-well deep well plate, one column was inoculated every other column, 1 mL of MRS liquid medium and 20 μL of bacterial liquid were added into each well, and each repeat was three plates. The bacterial body was collected by centrifugation at 3500 rpm for 8 min using a 96-well plate centrifuge, resuspended and washed with sterile water, repeated twice, and then resuspended with 550 μL of sterile water, mixed and used. Figure 1 Figure 2 The strain was streaked on the plate of MRS solid medium and activated, and incubated at 37°C in an incubator for 48 h. A smooth single colony was picked from the first generation plate and inoculated into a flask containing 30 mL of MRS liquid medium, and after 24 h of incubation at 37°C, a second generation seed liquid was obtained, and the viable cell count reached 10 8 -10 9 CFU / mL. The strain was added into each well of a 96-well deep well plate, one column was inoculated every other column, 1 mL of MRS liquid medium and 20 μL of bacterial liquid were added into each well, and each repeat was three plates. The bacterial body was collected by centrifugation at 3500 rpm for 8 min using a 96-well plate centrifuge, resuspended and washed with sterile water, repeated twice, and then resuspended with 550 μL of sterile water, mixed and used.

[0054] Example 2

[0055] 1 mL of bacterial liquid was centrifuged to remove the supernatant, the bacterial body was washed with physiological saline for 2 times, and 360 μL of PBS solution was used for resuspension to obtain bacterial suspension.

[0056] Glycine and taurine were generated after the decomposition of sodium glycocholate and sodium taurocholate, respectively. To two 180 μL bacterial suspensions, 20 μL of sodium glycocholate solution and sodium taurocholate solution (both 200 mM, solvent water) were added, respectively, and reacted at 37°C for 3 h. The degradation supernatant was collected by centrifugation. Glycine and taurine solutions with concentrations of 0.1, 0.2, 0.5, 0.8, 1.0, 2.0, 4.0 and 5.0 mM were prepared, respectively, and the absorbance of the two solutions at different concentrations was detected to establish the standard curve of glycine and taurine. According to the requirements of the kit, a liquid containing 20 μL of degradation supernatant was prepared, and the liquid was boiled in a water bath for 15 min. 200 μL was taken and used to detect the absorbance at 570 nm using the amino acid detection kit AA-W96-N (1620) to determine the concentration of sodium glycocholate and sodium taurocholate.

[0057] In addition, 360 μL of the above cholesterol solution (180 μL of sodium glycocholate solution and 180 μL of sodium taurocholate solution mixed) was heated to 37°C, and 30 mL of sterilized MRS liquid medium containing the rotor was added, and stirred at 750 rpm for 15 min. To 180 μL of bacterial suspension, 20 μL of the stirred sample was added, mixed, and incubated at 37°C for 15 min. The absorbance at 500 nm was detected using the cholesterol detection kit TC-W48-N (1031). 样品 In addition, the bacterial suspension was replaced with an equal amount of blank medium without the strain to set up a blank control group, and the absorbance A 空白对照 was measured.

[0058] The cholesterol degradation rate was calculated according to the following formula:

[0059] Cholesterol degradation rate = (A 空白对照 -A 样品 ) / A 空白对照 x 100%;

[0060] The average cholesterol degradation rates of CCNH411 and LGG were 49.5% and 38.3%, respectively.

[0061] Example 3

[0062] Streptococcus mutans was inoculated in LB liquid medium, anaerobically cultured and activated at 37°C for 18 h, and then transferred to the corresponding fresh culture medium. The culture was incubated until the viable bacterial count reached 10 8 CFU / mL, and then diluted with LB liquid medium to 10 5 CFU / mL as an indicator bacterial solution.

[0063] The viable bacterial count was 1 x 10 90.6 mL of activated bacteria solution of Lactobacillus paracasei CCNH411 and LGG at 109CFU / mL was inoculated into 30 mL of fresh MRS liquid medium, respectively, and after 18 h of static culture at 37°C, the supernatant was centrifuged and membrane treatment (membrane pore size is 0.22 μm) was performed.

[0064] Three groups of experimental groups were set: S1: 100 μL of supernatant, 100 μL of indicator bacteria solution; S2: 50 μL of supernatant, 150 μL of indicator bacteria solution; S3: 25 μL of supernatant, 175 μL of indicator bacteria solution; in addition, the supernatant was replaced with an equal volume of MRS liquid medium as a negative control group. The OD 600 values of the experimental groups and the negative control group were measured, and the inhibition rate (unit: %) was calculated according to the following formula, and the results are shown in Table 1:

[0065] Inhibition rate = (A0-A) / A0x100%.

[0066] Table 1

[0067]

[0068] The data in Table 1 show that the inhibition rate of Lactobacillus paracasei CCNH411 on Streptococcus mutans is always higher than that of LGG.

[0069] Example 4

[0070] 5 mL of Arla Crème Fraiche was added to each well of a 12-well plate, and the seed solution of CCNH411 and Lactobacillus paracasei CICC 25261 was taken by a pipette according to the inoculation amount of 10 vol%, and was added to the Crème Fraiche in the 12-well plate and was blown evenly, and was placed in a constant temperature condition of 16°C for 48 h, and then the pH value of each well of the two strains was measured by a pH instrument, and the average was 4.21 and 4.43, respectively, indicating that the acid production capacity of CCNH411 provided by the application is stronger.

[0071] The fermented Crème Fraiche obtained was stirred at 600 rpm, and when a large amount of water-like material (buttermilk) was precipitated and formed into a 2 cm diameter butter grain fat ball, the stirring was stopped. The buttermilk was removed by filtration, washed 3 times in cold water at 4°C to remove residual buttermilk, and fermented butter was obtained. The washed fermented butter was squeezed to remove the water therein, and at the same time, the butter grain was changed into a dense butter layer, and the water molecules were uniformly distributed in the butter in a small state.

[0072] 20 people with certain experience in sensory evaluation in the field were employed as evaluators, and the fermented butter obtained by CCNH411 and CICC 25261 was subjected to sensory evaluation according to the evaluation standard in Table 2, and the scores are shown in Table 3.

[0073] Table 2

[0074]

[0075] Table 3

[0076]

[0077] The total scores of CCNH411 and CICC 25261 are 32 and 29 respectively, wherein the off-flavor and sweet flavor scores of the two are the same, and the scores of other items of CCNH411 are higher than those of CICC 25261, which indicates that the butter fermented by CCNH411 provided by the application can obtain butter with more excellent flavor.

[0078] Example 5

[0079] The toothpaste was prepared according to the ingredients and proportions in Table 4, and the balance was deionized water.

[0080] Table 4

[0081]

[0082] The preparation process of CCNH411 bacterial powder is as follows: CCNH411 strain seed liquid is inoculated into a triangular flask with added liquid MRS medium at an inoculation amount of 2vol%, and is placed in a 37℃ anaerobic incubator overnight to obtain a fermentation liquid; the fermentation liquid is collected into a centrifuge tube, centrifuged at 6000rpm at 4℃ for 10min, and the supernatant is discarded; 0.9wt% physiological saline is added to the volume before centrifugation, and the bacterial slurry is harvested by repeating centrifugation for 3 times, and the bacterial slurry is suspended in 0.9wt% physiological saline to make the final volume 1 / 10 of the volume of the fermentation liquid to prepare a bacterial suspension. The bacterial suspension is mixed uniformly with skimmed milk powder at a volume ratio of 1:1, and then is divided into freeze-drying bottles and pre-frozen in a-80℃ refrigerator for 24h. After pre-cooling in a quick-freezing dryer for 1h, the pre-frozen sample is quickly placed in a material tray, and the sample should not be remelted during the process. After covering the quick-freezing dryer, vacuumization is started, and vacuum quick-freezing drying is carried out for 24h; the bacterial powder after freeze-drying is taken out and stored in a dry place for standby.

[0083] Sorbitol, xanthan gum and sodium pyrophosphate are dissolved in an appropriate amount of water in advance, and CMC is dispersed in glycerol under high-speed stirring for 5min. Deionized water is added to the paste machine, calcium hydrogen phosphate, sodium benzoate and polyethylene glycol solution are added, and high-speed stirring is carried out for 10-15min. Silicon dioxide, sodium dodecyl sulfate and CCNH411 bacterial powder are added, and high-speed stirring is carried out under vacuum for 2min. The essence is added, and high-speed stirring is carried out under vacuum for 20min. The pressure is-0.095MPa (gauge pressure) when discharging, and the paste is packaged with a composite hose.

[0084] 60 patients with periodontal disease aged 35-50 years old, half male and half female, who had not taken antibiotics or hormones in the past 3 months and had no systemic diseases were selected. The male and female patients were randomly divided into a control group and a test group, with 30 people in each group. The control group brushed their teeth with ordinary sample toothpaste in the morning and evening every day for 1 month, and the test group brushed their teeth with test sample toothpaste in the morning and evening every day for 1 month. The subjects were required to use a unified toothbrush, and the length of toothpaste used each time should cover the toothbrush head, 2 minutes each time, for 1 month. The effects on common symptoms of periodontal disease were observed.

[0085] The therapeutic effects were evaluated as three levels: significant effect, effect and no effect. Significant effect means that after treatment, the symptoms such as gum bleeding, swelling, pain and the like disappear, and the texture is firm; effect means that after treatment, the symptoms such as gum bleeding, swelling, pain and the like are greatly improved or there is a small amount of bleeding, and the texture is somewhat loose; and no effect means that after treatment, the gum bleeding, swelling, pain and the like are not improved. The effect data of the control group and the test group are shown in Table 5. It can be found that brushing teeth with the toothpaste prepared from the strain CCNH411 provided by the application can significantly relieve the symptoms caused by periodontal disease.

[0086] Table 5

[0087]

[0088] Preparation Example 1

[0089] The navel oranges were washed with tap water and drained. A part of the navel oranges was pulped to obtain whole fruit pulp. Another part of the navel oranges was peeled and juiced to obtain juice. Then, the whole fruit juice was obtained by mixing the whole fruit pulp, the juice and water in a weight ratio of 2:8:5. The navel orange whole fruit juice was filtered with nylon gauze, homogenized at 30 MPa for 6 min, adjusted to pH 4.5 with NaHCO3, and formulated by adding 8 wt% of high fructose syrup, and then pasteurized at 80°C for 10 min. 7 The CCNH411 was inoculated into 100 mL of navel orange whole fruit juice at an inoculation amount of 3 vol% and fermented at 35°C for 24 h to obtain probiotic fermented juice.

[0090] Preparation Example 2

[0091] The CCNH411 was cultured in liquid MRS medium at 30°C for 12 h, and then centrifuged at 4°C and 5000 rpm for 15 min. The bacteria were washed once with sterile normal saline and then centrifuged. The collected bacteria were diluted with sterile normal saline.

[0092] The 3rd grade lean meat is cut into pieces, and the marinating ingredients are added while stirring, and marinated for 5-8 hours at 0-2℃. The marinated meat is chopped, and fat, spices and starter are added and stirred evenly. The temperature of the meat during stirring is not higher than 2℃. The meat is stuffed into a casing, and the temperature of the meat during stuffing is 2-5℃. The strain 10 7 After fermentation at 10

[0093] Preparation Example 3

[0094] The probiotic ice cream is prepared according to the formula of whole milk powder (12wt%), white sugar (8wt%), butter (9wt%), malt dextrin (2wt%), coconut oil (4wt%), and the balance is pure water. The raw materials are mixed, sterilized at 85℃ for 15min, homogenized at 75-80℃ and 20MPa, and an aging slurry is obtained. The strain seed liquid is centrifuged at 10000rpm for 10min in a high-speed refrigerated centrifuge, and the bacterial suspension is collected. The bacterial suspension is inoculated into the aging slurry at 4℃ according to 10 7 CFU / mL, and stirred thoroughly until evenly distributed. The conventional freezing process is performed to obtain the ice cream product.

[0095] The above detailed the preferred embodiments of the present application, but the present application is not limited thereto. Within the technical concept of the present application, various simple modifications can be made to the technical solutions of the present application, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be considered as disclosed by the present application, and all fall within the protection scope of the present application.

Claims

1. A Lactobacillus paracasei strain Lactobacillus paracasei characterized in that, The preservation number of the Lactobacillus paracasei is CGMCC No. 33513.

2. An inoculant characterized in that, The bacterial agent contains the Lactobacillus paracasei of claim 1.

3. The microbial agent of claim 2, wherein, The bacterial agent further contains a supplementary material selected from glycerol and / or corn oil.

4. The microbial agent of claim 2, wherein, The number of viable L. paracasei is 10 4 -10 12 CFU per 1 mL of excipient.

5. Use of the Lactobacillus paracasei of claim 1 in the preparation of a preparation for inhibiting the growth and reproduction of Streptococcus mutans.

6. Use of the Lactobacillus paracasei of claim 1 in the preparation of butter by low-temperature fermentation.

7. Use according to claim 6, wherein, The amount of the Lactobacillus paracasei is such that the number of viable bacteria of Lactobacillus paracasei in the fermented raw material is not less than 10 6 CFU / mL.

8. The use according to claim 6, wherein, The conditions of the fermentation include a temperature of 15-35℃ and a time of 20-100h.

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