Lactobacillus paracasei and application thereof

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a lactobacillus paracasei and application thereof. The lactobacillus paracasei ZY101 is preserved in the China General Microbiological Culture Collection Center, has a preservation number of CGMCC No. 32360, a preservation date of October 25, 2024, and an address of No. 3, Beichen West Road, Chaoyang District, Beijing. Compared with a commercial bacillus acidophilus, the apricot vinegar prepared by using the lactobacillus paracasei ZY101 has new substances including linalool, alpha-farnesene, 2-phenylacetic acid ethyl ester and geranylgeraniol, and further promotes the aroma of the apricot vinegar. Compared with the commercial bacillus acidophilus, the existing substances are obviously improved, and specifically include ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl hexanoate, acetoin, methyl palmitate, gamma-deca-lactone, ethyl dodecanoate, delta-deca-lactone, trans-nerolidol and dihydro-beta-ionone.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine and relates to a strain of Lactobacillus paracasei and application thereof. BACKGROUND

[0002] Lactobacillus paracasei has been named as Lactobacillus paracasei (Lactobacillus paracasei) Lactobacillus paracasei ), belongs to the Lactobacillus casei group (LCG), and is one of the most valuable strains in lactic acid bacteria; Lactobacillus paracasei is a probiotic with the functions of enhancing immunity, maintaining intestinal flora balance, promoting digestion, reducing blood pressure, reducing cholesterol, etc., since 2008, Lactobacillus paracasei has been widely applied in the fields of dairy products, health foods, beverages, biscuits, etc.

[0003] Lactobacillus paracasei possesses potential probiotic properties, such as its acid and bile salt tolerance [PLESSAS S, NOUSKA C, KARAPETSAS A, et al. Isolation, characterization and evaluation of the probiotic potential of a novel Lactobacillus strain isolated from Feta-type cheese [J]. Food Chemistry, 2017, 226: 102-8.], and maintains good activity in simulated digestive fluids [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]. Furthermore, Lactobacillus paracasei plays a beneficial regulatory role in inflammation-related diseases. Studies have shown that *Lactobacillus paracasei* L9 improves colitis by expanding butyrate-producing bacteria that inhibit the IL-6 / STAT3 signaling pathway [J]. Food & Function, 2021, 12(21): 10700-13], and that *Lactobacillus paracasei* can alleviate colitis by regulating gut microbiota, protecting the intestinal epithelial barrier, reducing the content of inflammatory factors in the gut, and regulating the expression of host inflammation-related pathways. Furthermore, intranasal administration of *Lactobacillus paracasei* can reduce lung inflammation in mice with allergic bronchitis, thereby alleviating allergy symptoms [Ren, Rairway Inflammation in Mice [J]. International Journal of Inflammation, 2012, 2012.].In the atopic dermatitis mouse model, Lactobacillus paracasei can alleviate the clinical symptoms of the disease by reducing the level of mouse cell inflammation and changing the metabolism of intestinal microorganisms [KIM W-K, JANG Y J, HAN D H, et al. Lactobacillus paracasei KBL382 administration attenuates atopic dermatitis by modulating immune response and gut microbiota [J]. Gut Microbes, 2020, 12(1): 1819156.].

[0004] The vinegar industry in China uses Acetobacter during brewing, but the aroma substances produced by itself are difficult to meet market requirements. The metabolic products of Lactobacillus paracasei can produce "pleasant fruit aroma and cheese aroma", and Lactobacillus paracasei as a recognized probiotic can produce gamma-aminobutyric acid, hyaluronic acid, 3-hydroxybutyric acid and other beneficial products to the human body and can reduce blood pressure and blood lipids, and has a certain health care effect. SUMMARY

[0005] In view of the problem of single taste of vinegar varieties at the present stage, the present application provides a Lactobacillus paracasei and its application in preparing vinegar.

[0006] The scheme of the present application is as follows:

[0007] The first object of the present application is to protect a Lactobacillus paracasei, which is ZY101 deposited in the China General Microbiological Culture Collection Center; the deposit number is CGMCC No. 32360; the deposit date is October 25, 2024; and the deposit address is No. 3, Beichen West Road, Chaoyang District, Beijing.

[0008] The second object of the present application is to protect the application of the above-mentioned Lactobacillus paracasei in preparing vinegar.

[0009] Preferably, the application in apricot vinegar.

[0010] Preservation certificate of Lactobacillus paracasei ZY101:

[0011] Preservation agency: China General Microbiological Culture Collection Center (CGMCC);

[0012] Taxonomic name: Lactobacillus paracasei Lactobacillus paracasei ;

[0013] Deposit number: CGMCC No. 32360;

[0014] Deposit date: October 25, 2024;

[0015] Deposit address: No. 3, Xili Jiayuan, Beichen West Road, Chaoyang District, Beijing.

[0016] Advantages of the present application

[0017] Compared with the apricot vinegar prepared by using the commercial B. pasteurii, the newly added substances of the apricot vinegar prepared by using the L. paracasei ZY101 include linalool, alpha-farnesene, 2-phenylacetic acid ethyl ester, geranylgeraniol; and the aroma of the apricot vinegar is further promoted.

[0018] Compared with the commercial B. pasteurii, the existing substances are obviously improved, and specifically include ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl hexanoate, acetoin, methyl palmitate, gamma-decalactone, ethyl dodecanoate, delta-decalactone, trans-nerolidol, dihydro-beta-ionone. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 The colony morphology of part of the colonies separated from the persimmon sample in Example 1 on the plate;

[0020] Figure 2 The gram staining results of part of the colonies separated in Example 1. DETAILED DESCRIPTION

[0021] In order to better understand the present application, the content of the present application is further illustrated below in combination with examples, but the content of the present application is not limited to only the following examples. The materials, reagents and the like used in the examples and test examples of the present application are all conventional products obtained from commercial channels unless otherwise specified; and the methods used in the examples and test examples of the present application are all conventional methods unless otherwise specified.

[0022] Test Example 1

[0023] The obtaining steps of the L. paracasei ZY101 are specifically as follows:

[0024] Take 200g sample from persimmon in Yuncheng City of Shanxi Province, place it in a sealed bag, and ferment it naturally at 37°C for 3 days. After fermentation, take 50g sample and place it in a conical flask, and add 100ml distilled water. Place the conical flask in a shaker with conditions of 37°C and 220rpm for 18h to obtain a sample standard solution. Take 100ul of the sample standard solution and spread it on MRS solid culture medium, and incubate it at 37°C for 36h to obtain colonies. Select colonies with characteristics of Lactobacillus paracasei on the medium (milk yellow, round and raised, shiny, and transparent circle), and use an inoculation loop to pick colonies with large transparent circles on MRS solid culture medium, and incubate them at 37°C for 36h to obtain purified single colonies. Pick the purified single colonies and inoculate them into 5ml MRS liquid medium, and incubate them at 37°C for 24h to obtain bacterial liquid. Label each bacterial liquid and perform gram staining, and retain strains with typical characteristics of Lactobacillus paracasei to obtain Lactobacillus paracasei ZY101.

[0025]

[0026] Example 2

[0027] The preparation method of the Lactobacillus paracasei ZY101 fermentation liquor comprises the following steps:

[0028] Fermentation: inoculate the live Lactobacillus paracasei ZY101 into the fermentation medium at an inoculation amount of 1×10 7 CFU / g, and culture at 37°C for 36h, so as to obtain the fermentation liquor with a pH of 3.6;

[0029] The fermentation medium is composed of 5.224g of MRS medium and 100mL of water; the solution is sterilized at 121°C for 30min to obtain the sterilized liquor; and the liquor is cooled to 36°C to obtain the fermentation medium.

[0030] Experimental Example 3

[0031] Application of Lactobacillus paracasei ZY101 in preparation of apricot vinegar

[0032] 1. Experimental method

[0033] Select 1kg of fresh apricots of three varieties of Kate, Golden Kate and Golden Sun, and squeeze the apricots into juice, and divide the juice of each variety into two samples which are completely identical, and add 1g of pectinase to each sample to prepare apricot juice, and the apricot juice is divided into an experimental group and a control group;

[0034] Add 0.6g of activated active dry yeast (EC1118) (from Yantai Mansen Trade Co., Ltd.) to each sample, and perform fermentation at 17°C;

[0035] After 7 days of fermentation, add 33mL of the fermentation liquor of Lactobacillus paracasei ZY101 to the experimental group, and add 33mL of the activated commercial Bacillus acetacidifaciens SHOQIAN 1.01 (from Beijing Preserved Biological Technology Co., Ltd.) to the corresponding control group;

[0036] After acidification for 7 days at 30°C and 190r / min in a shaker, sample the control group and the experimental group for GC-MS and high-efficiency gas chromatography-mass spectrometry detection.

[0037] Organize volunteers to perform sensory evaluation of the apricot vinegar of the experimental group and the control group, score the flavor, acidity and taste, and count the results.

[0038] 2. Experimental results

[0039] After detection, the flavoring substances produced by the experimental group are more abundant than those of the control group, and the detection results are shown in Tables 1 and 2:

[0040] Table 1: GC-MS detection results of the experimental group

[0041]

[0042] Table 2 GC-MS detection results of the control group

[0043]

[0044] It was found that the experimental group was different from the control group in that:

[0045] The experimental group contained ingredients that the control group did not, including linalool, a-farnesene, 2-phenylacetic acid ethyl ester, geranylgeraniol;

[0046] The following substances in the experimental group were improved compared with the control group: ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl hexanoate, acetoin, methyl palmitate, gamma-decalactone, ethyl dodecanoate, delta-decalactone, trans-nerolidol, dihydro-beta-ionone.

[0047] As can be seen, the flavor substance species and content of the experimental group are higher than those of the control group, proving that the apricot vinegar prepared by ZY101 fermentation has higher value than the traditional method. New substances appear after fermentation: linalool, a-farnesene, 2-phenylacetic acid ethyl ester, geranylgeraniol; among them, linalool, a-farnesene, and geranylgeraniol can improve the taste and aroma of vinegar; in addition, linalool has medical and health care effects. The results of feeding linalool to mice showed that it had certain effects on the central nervous system, including sleep, anticonvulsant, and body temperature reduction, and increased with the dose. Linalool has the function of inhibiting the proliferation of various human lymphocyte leukemia cells, selectively inhibiting the growth and proliferation of human lymphocyte leukemia cells by improving the induction of cell apoptosis, and 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 used as an ideal high-efficiency low-toxicity anti-leukemia drug for development and use. (Chen Shangqi, Zhao Linghua, Xu Xiaojun. Natural linalool resources and their development and utilization [J]. Forestry Science and Technology Development, 2013, 27(02): 13-17.)

[0048] Acetoin, also known as 3-hydroxybutanone, is an important flavor substance in vinegar, and is a high-content volatile substance in vinegar after acetic acid and furfural. It is also the precursor of the 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. Related research on acetoin and lactic acid metabolism in bacillus pasteurii [D]. China University of Petroleum (East China), 2020).

[0049] Dihydro-beta-ionone has anti-cancer, anti-inflammatory, anti-oxidation, anti-trypanosoma, and anti-parasitic activities (Zhu F, 2023), and it can inhibit the proliferation of breast cancer cells by inhibiting the activity of NF-kappa B (Gao GJ, Wang FL, Li J, et al. Beta-ionone inhibits breast cancer cell proliferation through the NF-kappa B pathway. Practical Oncology, 2024, 38(04): 254-261).

[0050] Therefore, the apricot vinegar prepared by the Lactobacillus paracasei ZY101 strain can exert the above multiple uses through the effects of linalool, acetoin, and beta-ionone.

[0051] The sensory evaluation results show that most of the volunteers believe that the apricot vinegar prepared in the experimental group is stronger than the control group in terms of flavor, acidity, and taste. The specific results are shown in Tables 3 and 4. As can be seen from Tables 3 and 4, 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.

[0052] Table 3 Sensory evaluation score standard of apricot vinegar

[0053]

[0054] Table 4 Sensory evaluation summary of experimental group and control group

[0055]

Claims

1. A strain of Lactobacillus paracasei ( Lactobacillus paracasei ), characterized in that, The Lactobacillus paracasei ZY101 is preserved in the China General Microbiological Culture Collection Center; the preservation number is CGMCC No. 32360; the preservation date is October 25, 2024; and the preservation address is No. 1, Xibei Road, Beijing Chaoyang District, China.

2. Use of the Lactobacillus paracasei of claim 1 in the preparation of apricot vinegar.

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