WO2004090118A1 - Souche st-iii de lactobacillus plantarum et son utilisation pour la regulation de la lipidemie - Google Patents

Souche st-iii de lactobacillus plantarum et son utilisation pour la regulation de la lipidemie Download PDF

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Publication number
WO2004090118A1
WO2004090118A1 PCT/CN2003/000485 CN0300485W WO2004090118A1 WO 2004090118 A1 WO2004090118 A1 WO 2004090118A1 CN 0300485 W CN0300485 W CN 0300485W WO 2004090118 A1 WO2004090118 A1 WO 2004090118A1
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Prior art keywords
iii
group
cholesterol
content
bacteria
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PCT/CN2003/000485
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English (en)
Chinese (zh)
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WO2004090118A8 (fr
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Nengqun Jiang
Liguo Sun
Benheng Guo
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Shanghai Bright Dairy & Food Co., Ltd.
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Priority to AU2003252423A priority Critical patent/AU2003252423A1/en
Publication of WO2004090118A1 publication Critical patent/WO2004090118A1/fr
Publication of WO2004090118A8 publication Critical patent/WO2004090118A8/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

Definitions

  • Lactobacillus plantarum ST-III strain and its application in regulating blood lipids
  • the invention relates to a new strain of Lactobacillus and its application in regulating blood lipids. technical background
  • the present inventors have studied the microorganisms in the Chinese traditional food kimchi and pickled alfalfa as the target flora, and applied a high cholesterol culture medium to select a probiotic lactic acid bacteria having a cholesterol-lowering effect, and further studied its role in regulating blood lipids. Applications.
  • the object of the present invention is therefore to provide a Lactobacillus plantarum ST-III;
  • Another object of the present invention is to disclose the use of the aforementioned Lactobacillus plantarum ST-III in regulating blood lipids.
  • Lactobacillus plantarum ST-III of the present invention is as follows: refrigerate kimchi juice or alfalfa juice as a sample, measure its pH value, inoculate it on an MRS plate (purchased by Merck) and count the total colonies; Take the above sample, soak in PBS buffer solution with pH 6.8, mix and gradient dilute according to the above measurement results;
  • Lactobacillus plantarum ST-III of the present invention is used to regulate blood lipids, such as the development of functional dairy products, as a human dietary supplement, and to help reduce serum cholesterol in people Effect of concentration.
  • the invention will be explained in more detail by means of the following examples. The following examples are merely illustrative, and the present invention is not limited by these examples.
  • the Lactobacillus plantarum ST-III of the present invention was deposited on December 6, 2002 at No. 13, Zhongguancun North, Haidian District, Beijing, China. And received the deposit number CGMCC NO.0847. Summary of the Invention
  • Example 1 The samples were collected as follows: Kimchi juice: cabbage kimchi, vegetable paddles and juice homogenized together; alfalfa juice: Collected pickled alfalfa from Tianpingshan Farm in Suzhou, took 5 grams, and added 10 ml of sterile water to all The homogenate obtained in Example 2 was collected and screened with lactic acid bacteria. The above sample was filled into a sterile conical bottle, screwed tightly, and refrigerated at 4 ° C. The pH of kimchi juice and alfalfa juice was measured within 24 hours, and the total colony count was measured by inoculating the MRS plate, and then processed according to the following steps.
  • Example 3 Separate the kimchi juice and alfalfa juice from the LBS plate.
  • Example 2 use a PBS buffer solution (pH 6.8) to make a gradient dilution under aseptic conditions. Aspirate the appropriate dilution solution (100 ml per ml). left (Right bacteria) 100 microliters were coated on LBS plate. The plate was placed upside down in an anaerobic incubator and cultured at 37 C for 36-72 hours. The colony growth was observed. The plate was stored at 4 ° C.
  • the inoculating ring was used to pick up the typical colony culture obtained in Example 3, coated on a glass slide, prepared and stained with Gram, observed the morphology of the bacteria with an oil microscope, and picked Gram-positive bacteria as the target strain.
  • the inoculation loop picks the target colonies initially determined in Example 4, separates them on the MRS plate, and inverts anaerobic culture at 37 ° C for 24-36 hours. Picks single colonies for microscopic examination until the result is determined. The strain has been purified.
  • Example 5 Pick the ideal single colony obtained in Example 5, transfer it to 1 ml of MRS liquid, and leave it to stand at 37 ° C for about 24 hours. Then transfer the culture liquid to the new MRS liquid according to the inoculation amount of 1% (volume ratio). Incubate at 37 ° C for 24 hours, repeat the amplification in liquid MRS twice, and store the resulting culture solution at 4 ° C.
  • Example 6 A 15 ml sterile centrifuge tube was added with 10 ml of high-cholesterol culture medium MRSO-CHOL with sterile egg yolk as a cholesterol source, and the fresh strain obtained in Example 6 was inserted at a 1% (volume ratio) inoculation amount. Incubate at 37 ° C for 24 hours. The culture solution was centrifuged at 12,000 Xg for 10 minutes, and the cholesterol content in the supernatant was determined according to the method for measuring cholesterol by o-phthalaldehyde. Using uninoculated MRSO-CHOL as a control, the cholesterol reduction rate of each experimental strain was calculated (SEGlliland, CRNelson, C. Maxwell. Applied and Enviromental Microbiology, 1985, 49: 377-381.), From which strains with better cholesterol-lowering efficacy were selected. The results are shown in Table 2. The formula for reducing cholesterol is: c
  • A is the OD 55Qnm [absorption photometric value at a wavelength of 550nm (nm)] of the fermentation supernatant of each experimental strain;
  • ST-III has the highest cholesterol reduction rate, reaching 47.68%, and the corresponding cholesterol reduction amount is 244.84 ⁇ g / ml.
  • five strains had a cholesterol reduction rate of more than 40%.
  • the microbiological characteristics of the ST-III were tested and identified by the Institute of Microbiology, Chinese Academy of Sciences as shown in Table 1 above.
  • Lactobacillus plantarum ST-III (CGMCC No. 0847) of the present invention was put into MRS liquid, and 32 ° (rested for 16 hours, counted; according to the counting result, skim milk was appropriately diluted and suspended to obtain as shown in Table 3.
  • ST-III dead bacteria suspension preparation method Suspension 10 8 cfo / ml (the number of bacteria contained in each ml) ST-III skim milk is heated in a 100 ° C water bath for 15 minutes, cooled, Keep in the refrigerator for future use.
  • 0 high-fat high-cholesterol corpus induces animal lipid metabolism disorders, changes in blood lipid levels: serum cholesterol (TC) and triglyceride (TG) levels and low-density lipoprotein cholesterol (LDL-C) content Increased, high-density lipoprotein cholesterol (HDL-C) content decreased.
  • the normal method for lowering serum cholesterol is to feed high-fat and high-cholesterol feeds at the same time as feeding experimental animal experimental samples, and determine the effect of the experimental samples on animal lipids by measuring changes in animal serum lipid content.
  • the probiotic lactic acid bacteria screened by the present invention is Lactobacmus plantarum ST-III (CGMCC No. 0847), which has a high ability to reduce cholesterol concentration in in vitro experiments.
  • SD rats are used, and 1% cholesterol, 5% lard, and 0.25% pig bile salt are added to conventional feed to induce SD rats with hypercholesterolemia.
  • the skimmed milk of Lactobacillus plantarum was fed, and the serum cholesterol and other blood lipid contents of SD rats were finally measured to determine whether the strain had the effect of reducing serum cholesterol.
  • Conditions of animal room temperature 22 ⁇ 2 ° C, humidity 56 ⁇ 5%, change sawdust 2 to 3 times a week. Control 12 hours light and 12 hours darkness.
  • High Cholesterol Feed Conventional feed is added with a weight ratio of 1.0% cholesterol, 5% lard, and 0.25% pig bile salt, which is processed by the Shanghai Experimental Animal Center of the Chinese Academy of Sciences;
  • TC total serum Cholesterol
  • TG triglyceride
  • HDL-C high-density lipoprotein cholesterol
  • LDL-C low-density lipoprotein cholesterol
  • TAA total bile acid
  • Table 5 shows the HDL-C and LDL-C measurement results in experimental rats.
  • the difference has a high statistical significance a.
  • the experimental grouping is shown in Table 3; b AI-atherosclerosis risk factor, AI-LDL-C / HDL-C can be seen from Table 5, the high-fat control group The HDL-C content of 2 was 26.47% lower than that of the blank control group 1.
  • the difference was statistically significant ( ⁇ ⁇ 0.05), and the LDL-C content increased by 381.25%, and the difference was highly statistically significant (p ⁇ 0.001)
  • the atherosclerotic factor AI increased by 540.62%, and the difference was highly statistically significant (p ⁇ 0.001).
  • high-fat serving induces a disorder of lipoprotein metabolism in rats, a decrease in HDL-c content and LDL — The content of c increased, and the value of AI risk factors increased significantly.
  • the serum HDL-C and LDL-C contents of group 3 rats fed with SKM increased by 5.33% and decreased by 1.95% compared with the high-fat control group 2, respectively. The difference was not statistically significant (p> 0.05), and the AI factor was also not statistically significant. This indicates that SKM has no regulating effect on lipoprotein metabolism.
  • the serum HDL-C content and LDL-C content of group 4 rats were higher than those of high-fat SKM controls, respectively.
  • Group 3 increased by 13.92% and decreased by 23.84%, but the difference was not statistically significant (p> 0.05). This indicates that L. bulgaricus has a fat egg effect on rats No effect on the metabolism. C serum HDL- group of five rats were 32.91% higher than the group 3, the difference was significant (p ⁇ 0.05), the content of LDL-C was 31.12% lower than that in group 2, the difference was highly statistically significant (p ⁇ 0.01); and the AI factor was reduced by 45.08% (p ⁇ 0.001), and the difference was also highly statistically significant . This shows that L. acidophilus has the effect of increasing HDL-C and reducing LDL-C content in rats.

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  • Health & Medical Sciences (AREA)
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Abstract

La présente invention concerne une souche ST-III de Lactobacillus plantarum, en l'occurrence la N° 0847 selon CGMCC, et son utilisation pour la régulation de la lipidémie.
PCT/CN2003/000485 2003-04-11 2003-06-23 Souche st-iii de lactobacillus plantarum et son utilisation pour la regulation de la lipidemie WO2004090118A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2003252423A AU2003252423A1 (en) 2003-04-11 2003-06-23 Lactobacillus plantarum strain st-iii and its use in adjusting blood fat

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CNB031163777A CN1207382C (zh) 2003-04-11 2003-04-11 植物乳杆菌st－iii菌株及其在调节血脂方面的应用
CN03116377.7 2003-04-11

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WO2004090118A1 true WO2004090118A1 (fr) 2004-10-21
WO2004090118A8 WO2004090118A8 (fr) 2005-02-17

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107760620A (zh) * 2017-10-18 2018-03-06 天津科睿恒生物科技有限公司 一株改善易胖体质及子代体重的植物乳杆菌及应用

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CN100422309C (zh) * 2004-09-29 2008-10-01 光明乳业股份有限公司 植物乳杆菌st-iii的减肥用途
CN100337653C (zh) * 2004-12-21 2007-09-19 上海交大昂立股份有限公司 一种含有植物乳杆菌的组合物
CN102342327B (zh) * 2011-10-21 2013-01-02 上海光明奶酪黄油有限公司 含益生菌的切达干酪及其制备方法
CN102453767B (zh) * 2011-12-23 2013-07-17 光明乳业股份有限公司 一种快速定量检测植物乳杆菌st-ⅲ的方法及其试剂盒
CN102533790B (zh) * 2011-12-23 2013-10-02 光明乳业股份有限公司 一种检测植物乳杆菌st-ⅲ的pcr检测方法及其引物和试剂盒
CN102732470B (zh) * 2012-07-20 2014-01-08 光明乳业股份有限公司 一种纯天然、低成本的植物乳杆菌st-ⅲ培养方法及其产品和应用
CN103284028B (zh) * 2013-05-13 2014-12-17 陈正军 一种泡菜发酵剂及其制备方法和在蔬菜泡制中的应用方法
CN103598594B (zh) * 2013-10-28 2015-05-20 天津科技大学 植物乳杆菌在降血脂和辅助减肥方面的应用
CN104430851B (zh) * 2013-12-25 2017-08-25 广东燕塘乳业股份有限公司 一种降胆固醇发酵乳及其制备方法
CN105820989A (zh) * 2016-06-15 2016-08-03 苏州健世星生物科技有限公司 一种直投式发酵剂益生乳酸菌的高密度培养方法
CN109385389A (zh) * 2017-08-08 2019-02-26 光明乳业股份有限公司 一种缺失ldhA基因的植物乳杆菌ST-Ⅲ及其制备方法
CN107897368A (zh) * 2017-11-15 2018-04-13 光明乳业股份有限公司 一种益生菌发酵乳及其制备方法
CN108517308B (zh) * 2018-05-23 2021-06-04 光明乳业股份有限公司 一种微生态菌剂及其制备方法和应用
CN111004754B (zh) * 2019-12-30 2022-05-06 光明乳业股份有限公司 一种具有抗氧化乳酸菌制剂及其制备方法

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US5474932A (en) * 1991-07-25 1995-12-12 Probi Ab Intestine colonizing strains of lactobacilli
US6214336B1 (en) * 1997-08-05 2001-04-10 Probi Ab Use of lactobacillus for reduction of the fibrinogen level in blood

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5474932A (en) * 1991-07-25 1995-12-12 Probi Ab Intestine colonizing strains of lactobacilli
US6214336B1 (en) * 1997-08-05 2001-04-10 Probi Ab Use of lactobacillus for reduction of the fibrinogen level in blood

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107760620A (zh) * 2017-10-18 2018-03-06 天津科睿恒生物科技有限公司 一株改善易胖体质及子代体重的植物乳杆菌及应用
CN107760620B (zh) * 2017-10-18 2021-05-04 天津科睿恒生物科技有限公司 一株改善易胖体质及子代体重的植物乳杆菌及应用

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AU2003252423A1 (en) 2004-11-01
CN1207382C (zh) 2005-06-22
CN1467290A (zh) 2004-01-14
WO2004090118A8 (fr) 2005-02-17

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