Lactobacillus paracasei and application thereof

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to lactobacillus paracasei and application thereof. The lactobacillus paracasei ZY101 is preserved in the China General Microbiological Culture Collection Center (CGMCC); the preservation number is CGMCC No. 32360; the preservation date is October 25, 2024; the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Compared with commercial Acetobacter pasteurianus, the apricot vinegar prepared from the lactobacillus paracasei ZY101 has the advantages that newly added substances comprise linalool, alpha-farnesene, ethyl 2-phenylacetate and geranyl geraniol; the fragrance of the apricot vinegar is further promoted. Compared with commercial Acetobacter pasteurianus, the content of the existing substances is obviously increased, and the existing substances specifically comprise ethyl acetate, isoamyl acetate, ethyl caprylate, ethyl hexanoate, acetoin, methyl palmitate, gamma-decalactone, ethyl laurate, delta-decalactone, trans-nerol and dihydro-beta-ionone.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and relates to a strain of Lactobacillus paracasei and application thereof. Background Art

[0002] Lactobacillus paracasei was once named Lactobacillus paracasei ( Lactobacillus paracasei ), belongs to the Lactobacillus casei group (LCG), and is one of the most valuable species of lactic acid bacteria; Lactobacillus paracasei is a probiotic that has the effects of enhancing immunity, maintaining intestinal flora balance, promoting digestion, lowering blood pressure, and lowering cholesterol. Since 2008, Lactobacillus paracasei has been widely used in dairy products, health foods, beverages, biscuits and other fields.

[0003] Lactobacillus paracasei has potential probiotic properties, such as acid and bile resistance [PLESSAS S, NOUSKA C, KARAPETSAS A, et al. Isolation, characterization and evaluation of the probiotic potential of a novel Lactobacillus strainisolated from Feta-type cheese [J]. Food Chemistry, 2017, 226: 102-8.], and can maintain good activity in simulated digestive fluid [BENGOA AA, LLAMAS MG, IRAPORDA C, et al. Impact of growth temperature on exopolysaccharide production and probiotic properties of Lactobacillus paracasei strains isolated from kefir grains [J]. Food microbiology, 2018, 69: 212-8]. In addition, Lactobacillus paracasei plays a good regulatory role in inflammatory-related diseases. Studies have found that [DENG M, WU X, DUAN X, et al. Lactobacillus paracasei L9improves colitis by expanding butyrate-producing bacteria that inhibit theIL-6 / STAT3 signaling pathway [J]. Food&function, 2021, 12(21): 10700-13], Lactobacillus paracasei can alleviate colitis by regulating intestinal flora, protecting intestinal epithelial barrier, reducing the content of inflammatory factors in the intestine and regulating the expression of host inflammation-related pathways. In addition, Lactobacillus paracasei can reduce the pulmonary inflammatory response in mice with allergic tracheitis through nasal administration, thereby alleviating allergic symptoms [rairway inflammation in mice [J]. International journal of inflammation, 2012, 2012.].In a mouse model of atopic dermatitis, Lactobacillus paracasei can alleviate the clinical symptoms of the disease by reducing the level of cellular inflammation in mice and changing the metabolism of intestinal microorganisms [KIM WK, JANG YJ, HAN DH, et al. Lactobacillus paracaseiKBL382 administration attenuates atopic dermatitis by modulating immuneresponse and gut microbiota [J]. Gut Microbes, 2020, 12(1): 1819156.].

[0004] my country's vinegar industry uses acetic acid bacteria in brewing, but the flavor substances it produces are difficult to meet market demand. The metabolites of Lactobacillus paracasei can produce "pleasant fruity and cheese aromas", and as a recognized probiotic, Lactobacillus paracasei can produce γ-aminobutyric acid, hyaluronic acid, 3-hydroxybutyric acid and other products that are beneficial to the human body, and can lower blood pressure and blood lipids, playing a certain health care effect. Summary of the invention

[0005] In view of the problem that vinegar varieties currently have a single taste, the present invention provides a strain of Lactobacillus paracasei and application thereof in preparing vinegar.

[0006] The scheme of this application is as follows: The first purpose of the present invention is to protect a strain of Lactobacillus paracasei, wherein the Lactobacillus paracasei ZY101 is deposited in the General Microbiological Center of China Microorganism Culture Collection Committee; the deposit number is CGMCC No. 32360; the deposit date is October 25, 2024; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0007] The second object of the present invention is to protect the application of the above-mentioned Lactobacillus paracasei in preparing vinegar.

[0008] Preferably, the application is to apricot vinegar.

[0009] The preservation certificate of Lactobacillus paracasei ZY101: Depository: China General Microbiology Center (CGMCC); Taxonomic name: Lactobacillus paracasei Lactobacillus paracasei ; Deposit number: CGMCC No. 32360; Deposit date: October 25, 2024; Collection address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0010] Beneficial effects of the present invention Compared with commercial Acetobacter pasteurianus, the apricot vinegar prepared by using Lactobacillus paracasei ZY101 has newly added substances including linalool, α-farnesene, ethyl 2-phenylacetate, and geranylgeraniol, which further enhance the aroma of the apricot vinegar.

[0011] Compared with commercial Acetobacter pasteurianus, the existing substances are significantly improved, including ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl caproate, acetoin, methyl palmitate, γ-decalactone, ethyl dodecanoate, δ-decalactone, trans-nerol, and dihydro-β-ionone. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 The colony morphology of some colonies isolated from the persimmon sample on the plate in Example 1; Figure 2 The Gram staining results of some colonies isolated in Example 1 are shown. DETAILED DESCRIPTION

[0013] In order to better understand the present invention, the content of the present invention is further explained in conjunction with the following examples, but the content of the present invention is not limited to the following examples. The materials, reagents, etc. used in the examples and test examples of the present invention are conventional products obtained from commercial channels unless otherwise specified; the methods used in the examples and test examples of the present invention are conventional methods unless otherwise specified.

[0014] Experimental Example 1 The specific steps for obtaining Lactobacillus paracasei ZY101 are as follows: Taking persimmons from Hejin City, Yuncheng City, Shanxi Province as samples, 200 g of the sample was weighed and placed in a sealed bag, and naturally fermented at 37°C for 3 days. After the fermentation was completed, 50 g of the sample was weighed in a conical flask and 100 mL of distilled water was added. The conical flask was placed in a shaker at 37°C and 220 rpm and cultured for 18 hours to obtain the sample standard solution. 100 μL of sample standard solution was applied on MRS solid culture medium, and cultured at 37°C for 36 h to obtain colonies; colonies with characteristics of Lactobacillus paracasei on the culture medium (creamy yellow, round protrusions, glossy, and with transparent circles) were selected, and colonies with large transparent circles were picked up with an inoculation loop and streaked on MRS solid culture medium, and cultured at 37°C for 36 h to obtain purified single colonies; purified single colonies were picked up and inoculated into 5 mL of MRS liquid culture medium respectively, and cultured anaerobically at 37°C for 24 h to obtain bacterial liquid; each bacterial liquid was labeled and Gram-stained, and the strain with typical characteristics of Lactobacillus paracasei was retained to obtain Lactobacillus paracasei ZY101.

[0015]

[0016] Example 2 The preparation method of Lactobacillus paracasei ZY101 fermentation liquid comprises the following steps: Fermentation: The live bacteria of Lactobacillus paracasei ZY101 were added at a concentration of 1×10 7 CFU / g inoculum was inoculated into the fermentation medium, cultured at 37°C for 36 h, and the pH was measured to be 3.6 to obtain a fermentation broth; The fermentation medium consists of 5.224 g of MRS medium and 100 mL of water; the solution is sterilized at 121° C. for 30 min to obtain a sterilized feed solution; the feed solution is cooled to 36° C. to obtain a fermentation medium.

[0017] Experimental Example 3 Application of Lactobacillus paracasei ZY101 in the preparation of apricot vinegar 1. Experimental Methods Select 1 kg of fresh apricots of three varieties, Kate, Jin Kate and Jin Taiyang, and squeeze them into juice. The juice of each variety is divided into two identical samples and 1 g of pectinase is added to each sample and the juice is prepared by water bathing at 60℃ for 30 minutes. The apricot juice is divided into an experimental group and a control group. 0.6 g of activated active dry yeast (EC1118) (from Yantai Manson Trading Co., Ltd.) was added to each sample and fermented at 17°C; After 7 days of fermentation, 33 mL of fermentation liquid of Lactobacillus paracasei ZY101 was added to the experimental group, and 33 mL of activated commercial Acetobacter pasteurianus Huniang 1.01 (from Beijing Baozang Biotechnology Co., Ltd.) liquid was added to the control group; The control group and the experimental group were acidified in a shaker at 30°C and 190 r / min for 7 days and then sampled for GC-MS and high performance gas chromatography-mass spectrometry detection.

[0018] Volunteers were organized to conduct sensory evaluation of the apricot vinegar in the experimental group and the control group, and score its flavor, acidity, and taste and the results were counted.

[0019] 2. Experimental results After testing, the experimental group produced more aromatic substances than the control group. The test results are shown in Table 1 and Table 2: Table 1 GC-MS test results of the experimental group Table 2 GC-MS test results of the control group The test found that the difference between the experimental group and the control group was: The ingredients contained in the experimental group but not in the control group included: linalool, α-farnesene, ethyl 2-phenylacetate, geranylgeraniol; Compared with the control group, the contents of the following substances in the experimental group were increased: ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl caproate, acetoin, methyl palmitate, γ-decalactone, ethyl dodecanoate, δ-decalactone, trans-nerol, and dihydro-β-ionone.

[0020] It can be seen that the types and contents of flavor substances in the experimental group are higher than those in the control group, proving that apricot vinegar prepared by ZY101 fermentation has higher value than traditional methods. After fermentation, new substances appeared: linalool, α-farnesene, ethyl 2-phenylacetate, and geranylgeraniol; among them, linalool, α-farnesene, and geranylgeraniol can enhance the taste and aroma of vinegar; in addition, linalool has the effect of medical care. The results of the study on the administration of linalool to mice showed that it has certain effects on the central nervous system, including sleep, anticonvulsant, and hypothermia, and the effects increase with increasing doses. Linalool has the function of inhibiting the proliferation of a variety of human lymphocytic leukemia cells. It selectively inhibits the growth and proliferation of human lymphocytic leukemia cells by increasing the induction of cell apoptosis. It has no obvious effect on the proliferation of normal human bone marrow hematopoietic cells and peripheral blood cells. It has high safety for the human body and can be developed and used as a relatively ideal high-efficiency and low-toxic anti-leukemia drug. (Chen Shangying, Zhao Linghua, Xu Xiaojun. Natural linalool resources and their development and utilization [J]. Forestry Science and Technology Development, 2013, 27(02): 13-17.) Acetoin, also known as 3-hydroxybutanone (acetoin), is an important flavor substance in vinegar. It is a high-content volatile substance in vinegar after acetic acid and furfural. It is also the precursor of the iconic functional health ingredient ligustrazine. Ligustrazine has the effects of dilating blood vessels, increasing arterial blood flow, inhibiting platelet aggregation and reducing platelet activity. Therefore, increasing the content of acetoin and its heterocyclic derivative ligustrazine in vinegar is of great significance to improving the quality of vinegar (Ma Xiaolong. Research on acetoin and lactic acid metabolism in Acetobacter pasteurianus [D]. China University of Petroleum (East China), 2020).

[0021] Dihydro-β-ionone has multiple biological activities such as anti-cancer, anti-inflammatory, antioxidant, anti-trypanosomal, and anthelmintic (Zhu Xianxian, 2023), and it can inhibit the proliferation of breast cancer cells by inhibiting NF-κB activity (Gao Guangqiang, Wang Falin, Li Juan, et al. β-ionone inhibits breast cancer cell proliferation through NF-κB pathway [J]. Journal of Practical Oncology, 2024, 38(04): 254-261.).

[0022] Therefore, the apricot vinegar produced by the Lactobacillus paracasei ZY101 strain can play the above multiple purposes by producing linalool, acetoin and β-ionone.

[0023] The sensory evaluation results showed that the vast majority of volunteers believed that the apricot vinegar produced by the experimental group was stronger than that of the control group in terms of flavor, acidity and taste. The specific results are shown in Tables 3 and 4. It can be seen from Tables 3 and 4 that the taste and flavor of the apricot vinegar of the present application are significantly better than those of the control group, and the acidity is comparable to that of the control group.

[0024] Table 3 Sensory scoring standards for apricot vinegar Table 4 Summary of sensory evaluation of the experimental group and the control group

Claims

1. A strain of Lactobacillus paracasei, characterized in that: The Lactobacillus paracasei ZY101 is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration; the deposit number is CGMCC No. 32360; the deposit date is October 25, 2024; the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. application of Lactobacillus paracasei according to claim 1 in preparing vinegar.

3. The use according to claim 2, characterized in that: Application on apricot vinegar.

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