Search

CN-121991847-A - Lactobacillus helveticus (Lactobacillus helveticus) with blood lipid reducing effect, and sectional fermentation preparation method and application thereof

CN121991847ACN 121991847 ACN121991847 ACN 121991847ACN-121991847-A

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus helveticus (Lactobacillus helveticus) IOB726 with a blood lipid reducing effect, and a segmented fermentation preparation method and application thereof. The invention discovers that the Lactobacillus helveticus IOB726 has the efficacy of reducing blood fat for the first time. According to the invention, the lactobacillus helveticus IOB726 bacterial powder and the preparation thereof are prepared by the sectional liquid fermentation for the first time, so that not only is the bacterial activity improved, but also the inulin component is added into the second-stage fermentation culture medium, so that a carbon source is provided for the growth of the lactobacillus helveticus IOB726, short-chain fatty acid is promoted to be generated in the fermentation metabolism process of the lactobacillus helveticus IOB726, the synthesis of liver cholesterol is inhibited, and the hyperlipidemia symptom is further improved. In addition, the experiment result shows that the lactobacillus helveticus IOB726 has high-efficiency blood lipid reducing effect, has good acid and bile salt resistance and gastrointestinal tract tolerance simulation, can play a role in human intestinal tracts, supplements strain resources with blood lipid reducing probiotics, and shows better blood lipid reducing effect.

Inventors

  • Zong Dongsheng
  • LIANG WU
  • YU BAOCHEN
  • WANG HAIKUAN
  • HAN XUEMEI
  • YU ZEPING

Assignees

  • 沈阳药科大学
  • 沈阳血淤清中医药研发有限公司

Dates

Publication Date
20260508
Application Date
20260209

Claims (10)

  1. 1. The Lactobacillus helveticus (Lactobacillus helveticus) IOB726 with the blood lipid reducing effect is characterized in that the Lactobacillus helveticus is classified as Lactobacillus helveticus (Lactobacillus helveticus), the preservation number is CGMCC No.28044, the preservation time is 2023, 7 months and 28 days, the preservation place is China general microbiological culture Collection center (CGMCC), and the preservation unit address is North Star Xiyu No. 1, national academy of sciences of China, the Korean area of Beijing.
  2. 2. The Lactobacillus helveticus (Lactobacillus helveticus) IOB726 according to claim 1, wherein the Lactobacillus helveticus (Lactobacillus helveticus) IOB726 exerts an effect of improving hyperlipidemia symptoms by at least one of the following items (1) to (5) or a synergistic effect of improving hyperlipidemia symptoms: (1) Reducing cholesterol content in vitro; (2) Reducing triglyceride content in vitro; (3) Inhibiting pancreatic lipase activity; (4) Inhibiting cholesterol esterase activity; (5) Binding ability to cholate.
  3. 3. The method for preparing lactobacillus helveticus (Lactobacillus helveticus) IOB726 according to claim 1 or claim 2, wherein the method for preparing comprises the steps of: (1) Bacterial strain activation, namely taking lactobacillus helveticus (Lactobacillus helveticus) IOB726 out of a bacterial strain cryopreservation tube, inoculating the bacterial strain into an activation culture medium according to the volume ratio of 1:20-1:30, carrying out bacterial strain activation culture at the culture temperature of 36+/-2 ℃, sealing, standing for 24+/-2 hours, inoculating the activated bacterial strain which is the primary seed liquid into the activation culture medium according to the volume ratio of 1:30, and standing for 22+/-2 hours to obtain the secondary seed liquid; (2) And (3) sectional fermentation: The first stage of fermentation, namely inoculating the secondary seed liquid into a liquid fermentation culture medium according to the inoculum size with the volume ratio of 1:70-1:80, wherein the temperature is 36+/-2 ℃, the rotating speed is 120 r/min-140 r/min, and culturing for 18+/-2 hours to obtain a fermentation liquid; The second stage of fermentation, namely centrifuging the fermentation liquid of the first stage at 4 ℃ and 6000 r/min for 20min to concentrate the fermentation liquid, wherein the concentration multiple is 5-10 times, then inoculating the concentrated fermentation liquid into a second stage of liquid fermentation culture medium according to the volume ratio of 1:50-1:60, wherein the temperature is 36+/-2 ℃ and the rotating speed is 120 r/min-140 r/min, and culturing for 18+/-2 hours to obtain the fermentation liquid; (3) The preparation of the bacterial powder comprises the steps of centrifuging a fermentation liquid obtained after the second fermentation is finished at 4 ℃ and 4000 r/min for 15: 15min to respectively obtain supernatant and bacterial sludge, concentrating the supernatant, mixing the supernatant with the obtained bacterial sludge, precooling, performing vacuum freeze-drying, wherein the precooling temperature is-40 ℃, the vacuum degree is less than or equal to 10 Pa, and the freeze-drying time is 48+/-2 h, so that the Lactobacillus helveticus (Lactobacillus helveticus) IOB726 bacterial powder or a preparation thereof can be obtained.
  4. 4. A method of preparation according to claim 3, characterized in that: The activating culture medium is MRS culture medium, and the formula of the activating culture medium comprises peptone 8 g-10 g, yeast extract 3g-4 g, glucose 15 g-20 g, dipotassium hydrogen phosphate 1 g-2 g, triammonium citrate 1 g-2 g, mgSO 4 ·7H 2 O 0.2 g-0.5 g,MnSO 4 0.05 g-0.1 g, agar 8 g-10 g, tween 80.8 g-1 g, pH=6.0+ -0.2, and sterilizing 15min at 121 ℃ to 1L; The first-stage liquid fermentation culture medium is an improved MRS culture medium, and the formula of the first-stage liquid fermentation culture medium comprises 40 g-50 g parts of peptone, 15 g-20 g parts of yeast extract, 60-g-80 g parts of glucose, 5-g-10 g parts of dipotassium hydrogen phosphate, 5-g-10 g of tri-ammonium citrate, 0.25-g-0.5 g parts of MgSO 4 ·7H 2 O 1 g-2 g,MnSO 4 , 4-5 g of Tween 80, and pH=6.0+/-0.2 parts of yeast extract, wherein the volume is fixed to 5L, and the sterilization is carried out at 121 ℃ for 15min; The second-stage liquid fermentation culture medium is an improved MRS culture medium, and the formula of the second-stage liquid fermentation culture medium comprises 40 g-50 g parts of peptone, 15 g-20 g parts of yeast extract, 30 g-50 g parts of inulin, 5-g-10 g parts of dipotassium hydrogen phosphate, 5-g-10 g of tri-ammonium citrate, 0.25-g-0.5 g parts of MgSO 4 ·7H 2 O 1 g-2 g,MnSO 4 , 4-5 g of Tween 80, pH=6.0+/-0.2 parts of pH, and sterilizing at a constant volume of 5L and 121 ℃ for 15-min parts.
  5. 5. A product with hypolipidemic effect, characterized in that it comprises viable and/or inactivated cells of Lactobacillus helveticus (Lactobacillus helveticus) IOB726 according to claim 1 or claim 2.
  6. 6. Use of lactobacillus helveticus (Lactobacillus helveticus) IOB726 according to claim 1 or claim 2, lactobacillus helveticus (Lactobacillus helveticus) IOB726 powder or a preparation thereof prepared by the preparation method according to claim 3 or claim 4 or a product according to claim 5 for preparing a product with hypolipidemic effect.
  7. 7. The method according to claim 6, wherein the product is selected from the group consisting of a pharmaceutical, a functional food and a health food.
  8. 8. The method of claim 6, wherein the product is in an oral dosage form.
  9. 9. The method according to claim 8, wherein the oral dosage form is a tablet, capsule, oral liquid, enteric-coated tablet, granule, syrup or sublingual tablet.
  10. 10. The method of claim 6, wherein the product further comprises other drugs, functional foods or health products for preventing, improving or treating hyperlipidemia.

Description

Lactobacillus helveticus (Lactobacillus helveticus) with blood lipid reducing effect, and sectional fermentation preparation method and application thereof Technical Field The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus helveticus (Lactobacillus helveticus) IOB726 with a blood lipid reducing effect, and a segmented fermentation preparation method and application thereof. Background Hyperlipidemia is a common metabolic disease, mainly caused by abnormal elevation of lipid components such as cholesterol and triglyceride in blood, and may cause serious consequences such as arteriosclerosis, cardiovascular diseases, cerebrovascular diseases and the like. Hyperlipidemia is one of the important causative factors of atherosclerotic cardiovascular disease, and has become a serious problem in the public health field. Therefore, prevention and treatment of hyperlipidemia are particularly important. With the improvement of the living standard of people, the acceleration of the living rhythm, the increase of the living pressure, the reduction of the physical labor and the movement quantity and the deterioration of the work and the environment, the reduction and the disorder of the metabolic capability of human bodies, the reduction of the immunity and the continuous increase of the proportion of sub-healthy people are easily caused. In particular, the diet is greatly changed, and the high-fat and steroid foods are of a specific nature in daily life. However, long-term, excessive consumption of such foods can promote the occurrence of chronic diseases including hyperlipidemia, which can cause abnormal lipid metabolism of the body, increase of cholesterol and/or triglyceride in serum, seriously affect normal life and endanger health. At present, the clinical products in the aspect of blood lipid regulation are mainly made of medicines, such as fibrates, statins, nicotinic acid medicines and the like, which are effective in reducing blood lipid in one aspect, but have the defects of (1) large side effect, easy liver enzyme abnormality, gastrointestinal discomfort, abdominal pain and diarrhea and anorexia, 2) short duration effect, usually after taking the medicines, a plurality of treatment courses for keeping the curative effect, poor duration of the effect, 3) unstable effect, such as complex ingredients of the compound traditional Chinese medicine, and unstable treatment mechanism, so that the curative effect is always different from person to person, and 4) high cost, wherein long-term taking of the medicines brings great economic pressure for patients. With the development of science and technology and the improvement of the living standard of people, more and more patients with hyperlipidemia hope to have products with more comprehensive functions, healthier and safer, and not just reduce the blood fat. Probiotics are active microorganisms that when ingested in sufficient amounts can produce health benefits. A large number of researches at home and abroad show that a part of probiotics have remarkable effects on reducing blood fat in serum. Although there is considerable understanding of the link between the microbial flora and specific diseases of the human body, particularly chronic diseases, to date there are very few lactic acid bacteria resources clinically used to ameliorate hyperlipidemia. Therefore, finding a lactic acid bacterium that can lower blood cholesterol and triglycerides is an important breakthrough in this field. Disclosure of Invention The invention aims to overcome the defects of the prior art and provides lactobacillus helveticus (Lactobacillus helveticus) IOB726 with a blood lipid reducing effect, and a segmented fermentation preparation method and application thereof. The lactobacillus helveticus disclosed by the invention can reduce the cholesterol and triglyceride content in blood, and inhibit the activity of pancreatic lipase and cholesterol esterase. In order to achieve the above object, the present invention adopts the following technical scheme: In a first aspect, the invention provides lactobacillus helveticus (Lactobacillus helveticus) IOB726 with a hypolipidemic effect, wherein the lactobacillus helveticus is classified under the name of lactobacillus helveticus (Lactobacillus helveticus), the preservation number is CGMCC No.28044, the preservation time is 28 days of 7 months in 2023, the preservation place is China general microbiological culture Collection center (CGMCC), and the preservation unit address is China center for China, the university of China, the North Star Xiu Hill 1, the Beijing area, and the national institute of microbiology. The lactobacillus helveticus (Lactobacillus helveticus) IOB726 is obtained by screening a traditional yogurt product by the inventor. Alternatively, lactobacillus helveticus (Lactobacillus helveticus) IOB726 of the present invention reduces cholesterol and triglyceride levels, inhibits