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Novel lactobacillus fermenti and application thereof in lactic acid bacterium drink

A technology for fermented lactobacillus and lactic acid bacteria beverages, which is applied to a new type of fermented lactobacillus and its application field in lactic acid bacteria beverages, and achieves the effects of increased quantity, strong tolerance and broad application prospects

Active Publication Date: 2017-02-22
JINAN KANGDUOBAO BIOTECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(4) must actively compete for nutrients in the host

Method used

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  • Novel lactobacillus fermenti and application thereof in lactic acid bacterium drink
  • Novel lactobacillus fermenti and application thereof in lactic acid bacterium drink

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Lactobacillus fermentum KDB-8 isolation screening and identification:

[0023] 1. Screening samples

[0024] Naturally fermented sauerkraut juice was collected from the countryside of Sujiatun, Shenyang as an isolated sample. The sauerkraut juice has no pollution in appearance, pure taste, and has been fermented for more than one year. It is filtered and used for later use.

[0025] 2. Preliminary screening of lactic acid bacteria

[0026] Take 5 mL of filtered sauerkraut juice and add 45 mL of sterile water, mix well, spread on the MRS plate with 0.5% calcium carbonate, incubate at 37°C for 48 hours, isolate and culture the strains with transparent circles, and obtain 42 strains of lactic acid bacteria.

[0027] 3. Screening of antibacterial lactic acid bacteria

[0028] 1) Preparation of lactic acid bacteria liquid: inoculate 32 strains of lactic acid bacteria obtained from primary screening into 100mL MRS liquid medium, and culture at 37°C for 48 hours...

Embodiment 2

[0037] Example 2 Lactobacillus fermentum KDB-8 acid resistance, bile salt resistance experiment

[0038] 1. Gastric acid resistance test method:

[0039] Get 40ml of activated Lactobacillus fermentum KDB-8 bacterial suspension and add it to a 50ml centrifuge tube, centrifuge (5000g, 5min) to collect the thalline, wash twice with PBS buffer, add 40ml artificial gastric juice (take dilute hydrochloric acid) to the centrifuge tube Add 16.4ml of water and shake to dilute to 1000ml, adjust the pH to 2.0 with concentrated hydrochloric acid or 10% NaOH, sterilize at 121°C for 30min, add pepsin at a ratio of 1g / 100ml in a sterile room, incubate in a water bath at 37°C for 2h, Shake once every 30 minutes, take samples at 0h and 2h for counting viable bacteria, and use the 0h sample as a control to calculate the survival rate of the 2h sample bacteria, survival rate = (the number of viable bacteria at 2h / the number of viable bacteria at 0h) ×100%.

[0040] 2. Test method for bile salt...

Embodiment 3

[0046] Embodiment 3 Lactobacillus fermentum KDB-8 is to the removal effect of cholesterol

[0047] This experiment refers to the method of Brashears et al. (1998) with slight improvements.

[0048] The activated Lactobacillus fermentum KDB-8 strain was inoculated in the MRS medium and cultured overnight at 37°C as the seed solution.

[0049] Prepare MRS medium containing 5% (v / v) egg yolk liquid, take 2ml of medium, centrifuge at 5000r / min for 7min, take 1ml of supernatant and put it in a centrifuge tube, add 9ml of absolute ethanol, shake for 5min, 10000r Centrifuge for 10 min at 1 / min, take 2 ml of supernatant and add 2 ml of P-Fe-S reagent, mix and react in ice bath for 30 min, measure OD550, and calculate the initial content of cholesterol in the medium.

[0050] Then, inoculate the seed liquid of Lactobacillus fermentum KDB-8 into the above-mentioned MRS medium, and after culturing at 37°C for 24 hours, measure the final content of cholesterol in the medium again, and ca...

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Abstract

The invention relates to the technical field of microbiological screening and application and specifically provides a novel lactobacillus fermenti strain. The lactobacillus fermenti strain has a collection number of CCTCC NO: M2016430. The strain has obvious inhibitory effects on Escherichia coli, salmonella and staphylococcus aureus, particularly has the strongest inhibitory effect on helicobacter pylori, and the diameter of an inhibition zone reaches 23mm. The lactobacillus fermenti is high in tolerance to gastric acid and bile salts, and is capable of realizing effective multiplication in a gastric acid environment. The lactobacillus fermenti has a cholesterol removal rate of 93.2%, and can maintain the survival rate of 98.5% in 1.2mmol / L H2O2; the fermented supernatant fluid and cell-free extract of the lactobacillus fermenti respectively have the hydroxyl radical scavenging rate of 95.1% and 90.4%. The lactobacillus fermenti can be widely applied to preparation of the lactic acid bacterium drink, is capable of obviously increasing the quantity of intestinal available bacteria, improving the gastrointestinal tract health and improving the immunity of the organism, and has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of microbial screening, and specifically provides a novel lactobacillus fermentum and its application in lactic acid bacteria beverages. Background technique [0002] The concept of probiotics comes from the Greek word "bacteria beneficial to life". Probiotics are usually the general term for active beneficial microorganisms (bacteria or yeast) that colonize in the intestinal tract and reproductive system of animals and can produce definite health effects. The FAO / WHO "Guidelines for the Evaluation of Food Probiotics" clearly stipulates that food probiotics refer to "living microorganisms that are beneficial to human health after taking a certain amount". The effectiveness of probiotics is closely related to the number of live bacteria and the performance of the strain itself. relevant. In my country, probiotics are also called microecological regulators, prebiotics, etc. Probiotic health food clearly stipu...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L2/38A23L33/135C12R1/225
CPCA23L2/38C12N1/20A23V2002/00C12R2001/225C12N1/205A23V2400/11A23V2200/32A23V2200/324
Inventor 尤升波郭学建刘凤彩张静孙玉亮岳寿松游银伟朱友峰邹晓凤
Owner JINAN KANGDUOBAO BIOTECH CO LTD
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