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CN-120118796-B - Lactococcus garvieae ZB15, cell-free fermentation freeze-dried product with anti-inflammatory activity and preparation method thereof

CN120118796BCN 120118796 BCN120118796 BCN 120118796BCN-120118796-B

Abstract

The invention relates to the technical field of fermentation, in particular to a lactococcus garvieae ZB15 and a cell-free fermentation freeze-dried product with anti-inflammatory activity and a preparation method thereof. The lactococcus garvieae (Lactococcus garvieae) ZB15 is preserved in the microorganism strain collection center of Guangdong province, the address is building 5 No. 59 of No. 100 university in Guangzhou martyr, the preservation date is 2025, 02 month and 27 days, and the preservation number is GDMCC No:65947. The lactococcus garvieae ZB15 is prepared into a cell-free fermentation freeze-dried product, and the cell-free fermentation freeze-dried product can relieve and improve enteritis and can improve the immunity of organisms by changing the intestinal microflora structure at the same time, thereby providing a basis for preparing enteritis medicaments.

Inventors

  • WANG LING
  • LI GUOJIN
  • LI JIAO
  • LIU QIAN
  • GAO JIE
  • LU HONGCHAO
  • WANG SHANSHAN
  • ZHANG TAO
  • ZENG WENXIAN

Assignees

  • 陕西理工大学

Dates

Publication Date
20260512
Application Date
20250325

Claims (8)

  1. 1. A strain of lactococcus garvieae (Lactococcus garvieae) ZB15 is deposited in the microorganism strain collection center of Guangdong province, and has the address of building 5 of No. 59 of Dai 100 in Guangzhou city martyr, the date of deposit is 2025, 02 month and 27 days, and the deposit number is GDMCC No: 65947.
  2. 2. Use of lactococcus garvieae (Lactococcus garvieae) ZB15 according to claim 1 for the preparation of a medicament for the treatment of colitis.
  3. 3. A cell-free fermented lyophilisate having anti-inflammatory activity, characterized in that it is obtained by fermenting, freeze-drying the lactococcus garvieae (Lactococcus garvieae) ZB15 of claim 1.
  4. 4. A method for preparing a cell-free fermented lyophilized product with anti-inflammatory activity, comprising the steps of: (1) Inoculating the lactococcus garvieae (Lactococcus garvieae) ZB15 of claim 1 into MRS solid culture medium, and culturing for 20-28 h at 36-38 ℃ to obtain single colony; (2) Inoculating the single colony in the step (1) into an MRS liquid culture medium, and culturing for 20-28 h at 36-38 ℃ to obtain seed liquid, and inoculating the seed liquid into the MRS liquid culture medium, and culturing for 20-28 h at 36-38 ℃ to obtain working bacterial liquid; (3) Inoculating the working bacterial liquid into an MRS broth culture medium, and culturing for 20-28 hours at 36-38 ℃ to obtain fermentation liquor; (4) Lyophilizing the cell-free fermentation supernatant to obtain a cell-free fermentation lyophilized product; the freeze-drying in the step (4) comprises the steps of precooling and vacuum freezing; the pre-cooling temperature is-75 to-85 ℃, and the pre-cooling time is 18-22 hours; the temperature of the vacuum freezing is-45 to-55 ℃, the vacuum degree of the vacuum freezing is 20-40 Pa, and the time of the vacuum freezing is 22-26 h.
  5. 5. The method of claim 4, wherein the working bacteria liquid in step (3) is inoculated in an amount of 0.5-1.5% by volume of the MRS broth.
  6. 6. The preparation method according to claim 4, wherein the rotational speed of the centrifugation in the step (3) is 3500-4500 rpm, and the centrifugation time is 15-25 min.
  7. 7. The method according to claim 4, wherein the filtration in the step (3) is a filtration with a pore size of 0.22 μm or less.
  8. 8. Use of the cell-free fermented lyophilisate of claim 3 or the cell-free fermented lyophilisate prepared by the preparation method of any one of claims 4 to 7 in the preparation of a medicament for treating colitis.

Description

Lactococcus garvieae ZB15, cell-free fermentation freeze-dried product with anti-inflammatory activity and preparation method thereof Technical Field The invention relates to the technical field of fermentation, in particular to a lactococcus garvieae ZB15 and a cell-free fermentation freeze-dried product with anti-inflammatory activity and a preparation method thereof. Background Ulcerative colitis (ulcerative colitis, UC) is a recurrent inflammatory bowel disease, one of the main types of inflammatory bowel disease, with the main characteristics of UC being manifested by reduced body mass, abdominal pain, diarrhea and hematochezia. Although UC is more common in western countries, the incidence of UC in developing countries has also increased dramatically in recent years, which has led to UC becoming one of the 21 st century global health problems. The occurrence of UC is closely related to a variety of factors, including genetic factors, environmental factors, immunomodulation, intestinal microorganisms, lifestyle, etc., the exact pathogenesis of which is still unknown, and no effective cure is currently found. Conventional UC intervention drugs include corticosteroids, 5-aminosalicylic acid, immunosuppressants, antibiotics and the like, but have limited effects and are accompanied by a series of side effects, and toxicity caused by long-term administration can limit the curative effect. Therefore, it is important to explore a safe and effective way of preventing or improving UC. Research shows that the human body intake probiotics has a certain inhibition effect on the enteritis. Lactic acid bacteria and bifidobacteria are widely present in humans as important members of probiotics. Both are physiological bacteria of human body, maintain the health of host intestinal tracts, and inhibit the occurrence of cancers to a certain extent. The anti-tumor function of bifidobacterium infantis (Bifidobacteriuminfantis, b.infantis) in mice was first reported in 1978, after which a number of in vivo experiments have proved that lactic acid bacteria-enriched fermented foods are beneficial for human body and can inhibit the occurrence of tumors. For example, long-term consumption of yogurt or fermented dairy products can reduce the incidence of proctitis or other cancers, while ingestion of bulgaricus yogurt can inhibit the growth of colon tumors. Studies of Rafter and Commane show that the possible ways of inhibiting colon cancer by lactic acid bacteria are as follows, changing the number or kind of intestinal microbiota of the host, binding or degrading potential carcinogens, producing anti-tumor or anti-mutagenic substances, inhibiting tumor growth, improving the immune stress of the host and influencing the physiological metabolism of the host, etc. The immune system plays a vital regulatory role in the occurrence and development process of body tumors, and researches show that the colorectal cancer resistance of the lactobacillus is closely related to the immune regulation of the lactobacillus. Studies show that lactobacillus can increase macrophage activity, activate NK cells of mice, promote human peripheral blood mononuclear cells to release tumor necrosis factors TNF-alpha, interleukin IL-10, IL-12 and the like, stimulate dendritic cells to release IL-12, and regulate and control the release of IL-10. This means that lactic acid bacteria can achieve antitumor function by improving immunity of the organism. Wang Shumei et al showed that Lactobacillus paracasei (Lactobacillusparacasei subsp.paracasei,L.paracasei subsp.paracasei)M5、L.coryniformis subsp.torquens T3、 Lactobacillus rhamnosus (L.rhamnosus) SB5, SB31, J5, live cell walls and cell DNA of INI promote Proliferation of Blood Mononuclear Cells (PBMCs) in vitro and promote release of IL-12, IFN-gamma and TNF-alpha, thereby achieving its immunopotentiating function. The research results show that the lactobacillus has certain probiotics effect on the organism, and especially can improve the immunity of the organism. At present, the occurrence mechanism of colonitis is not clear, and the toxic and side effects generated by drug treatment are not negligible. Lactic acid bacteria are accepted probiotics, the antibacterial and anti-inflammatory effects of which have been confirmed by a great deal of researches, and some researchers use lactic acid bacteria to intervene and treat colitis, but the potential of the lactic acid bacteria for treating colitis is not completely released, and a plurality of non-negligible defects exist. Such as differences in the composition, immune status and genetic background of the patient's intestinal flora, can lead to differences in the efficacy of the lactic acid bacteria from person to person, and the original flora in the patient's intestinal tract may inhibit the colonization or function of exogenous lactic acid bacteria. Some researches show that lactic acid bacteria can aggravate inflammation by activating TLR signal chann