German-style lactobacillus with oxidation resistance and bacteriocin producing capability

A technology of Lactobacillus germanus and anti-oxidation ability, which is applied in the direction of bacteria, microbe-based methods, microbiological measurement/testing, etc., can solve the problems of scarcity of probiotics and lack of natural functional lactic acid bacteria resources, etc., and achieve strong peptidase and glycoside The effect of enzymatic hydrolysis activity, strong comprehensive antioxidant capacity, and good probiotic properties

Inactive Publication Date: 2017-03-29
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, natural functional lactic acid bacteria resources are scarce, and probiotics with high antioxidant capacity and bacteriocin production are even more scarce

Method used

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  • German-style lactobacillus with oxidation resistance and bacteriocin producing capability
  • German-style lactobacillus with oxidation resistance and bacteriocin producing capability
  • German-style lactobacillus with oxidation resistance and bacteriocin producing capability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Isolation and identification of lactic acid bacteria from traditional fermented yak yogurt on the Qinghai-Tibet Plateau

[0037] 1. Sample source

[0038] Originated from traditional fermented yak dairy products in Tianzhu Tibetan Autonomous County, Gansu Province, my country.

[0039] 2. Lactic acid bacteria isolated from traditional yak dairy products

[0040] Take 1mL of fermented yak dairy products at a ratio of 1:10 and dilute with sterilized saline, which is 10 -1 Dilution. Then draw 1mL of the above diluent, and then dilute it 10 times to make 10 -2 Dilution, and so on. Select 10 -5 , 10 -6 , 10 -7 For three dilutions, draw 100μL of the dilutions and apply them to MRS, M17, KFS and SL (pH 5.4) plate medium, respectively, at 37℃, 42℃, 42℃ and 37℃ anaerobic constant temperature Cultivate for 48 hours, pick characteristic colonies, and further purify to obtain purely isolated lactic acid bacteria strains.

[0041] Three, identify the strain

[0042] (1) Gram stai...

Embodiment 2

[0048] Example 2: Screening of lactic acid bacteria with high antioxidant capacity

[0049] 1. Preparation of intact cells and cell-free extracts of lactic acid bacteria

[0050] Preparation of cell bacterial solution: Put the bacterial solution stored in the laboratory at -80°C into sterilized MRS medium and culture it overnight at 37°C, and pass it 3 consecutive times. The culture solution was centrifuged at 12,000g to collect the bacteria, washed with PBS 3 times, resuspended in PBS, and adjusted the number of bacteria.

[0051] Preparation of cell-free extract: Activate the bacterial solution overnight at 37°C twice, 8000g, 10min, 4°C, centrifuge to collect the bacteria, wash with PBS 3 times, resuspend in PBS, adjust the number of bacteria to 1×10 9 CFU / mL or 1×10 10 CFU / mL. Cells were disrupted by ultrasonic in ice bath, 5s, 5s, 30min, 360W. 8000g, 10min, 4°C, the supernatant was collected as a cell-free extract (Guo Xinghua, 2008).

[0052] Second, preliminary screening

[005...

Embodiment 3

[0076] Example 3: In vitro probiotic properties study

[0077] 1. Invented strain enzyme activity assay (API-ZYM)

[0078] The API-zym reagent of French Mérieux company is used for detection. This technology is a semi-quantitative micro-method system specially designed for studying enzyme activity. The bacteria solution was subcultured 3 times with sterile MRS culture solution, collected by centrifugation, and prepared 1×10 with sterile saline 7 CFU / mL bacterial suspension was tested.

[0079] Table 2 shows that Lactobacillus F17 has strong peptidase activity, can effectively hydrolyze amino acids into volatile molecules, and plays a very important role in the flavor production of fermented products. The strong glycosidase activity of F17 can effectively utilize the sugar source and determine the energy supply of lactic acid bacteria fermentation. The higher the galactosidase activity, the more effective the human intestinal lactose intolerance.

[0080] Table 2 Enzyme activity of la...

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Abstract

The invention discloses a strain of German-style lactobacillus which is separated from traditional fermented yak yoghurt in Tibetan Plateau and has oxidation resistance and bacteriocin producing capability. The preservation number is CGMCC No.11247, the activity of GSH-Px and T-SOD in mouse bodies can be remarkably improved, the content of MDA in serum of mice is decreased, and the German-style lactobacillus has the immunoregulation function and can be applied to preparation of fermented milk products and health-care food and medicine.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to the field of development and application of functional lactic acid bacteria in extreme environments, and the separation, screening, identification and probiotics of Lactobacillus germanica subsp. bulgaricus with antioxidant function, bacteriocin-producing ability, and spoilage inhibition effect Functional evaluation, and application in food and pharmaceutical production. Background technique [0002] Oxidation is a necessary metabolic process of organisms, but excessive oxidation can cause damage to biological macromolecules. Various reactive oxygen molecules (ROS) are continuously produced in the process of oxidative metabolism. ROS have high oxidative activity. They are extremely unstable and extremely active. They can attack cell membranes and mitochondrial membranes, and react with unsaturated fatty acids in the membranes to cause lipids. Peroxidation is enhanced and more ROS...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C12R1/225
Inventor 郭旭生丁武蓉柯文灿陈明
Owner LANZHOU UNIVERSITY
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