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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  • Abstract
  • Description
  • Claims
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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

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[0022] Example 1 Isolation, screening and identification of Lactobacillus fermentum KDB-8:

[0023] 1. Screen samples

[0024] Naturally fermented sauerkraut juice was collected from the rural area of ​​Sujiatun, Shenyang as a separation sample. The sauerkraut juice has no pollution in appearance and pure taste, and the fermentation time exceeds 1 year, and it is filtered for use.

[0025] 2. Initial screening of lactic acid bacteria

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

[0027] 3. Screening of antibacterial lactic acid bacteria

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

[0029] 2) Prepara...

Example Embodiment

[0037] Example 2 Acid resistance and bile salt resistance experiment of Lactobacillus fermentum KDB-8

[0038] 1. Test method for gastric acid resistance:

[0039] Take 40ml of activated Lactobacillus fermentum KDB-8 bacterial suspension into a 50ml centrifuge tube, centrifuge (5000g, 5min) to collect the bacteria, wash twice with PBS buffer, add 40ml artificial gastric juice to the centrifuge tube (take dilute hydrochloric acid) Add 16.4ml with water and 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, each Shake well every 30 minutes, take samples at 0h and 2h to count the viable bacteria. Use the 0h sample as the control to calculate the survival rate of the 2h sample. Survival rate = (the number of viable bacteria at 2h / the number of viable bacteria at 0h) ×100%.

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

[00...

Example Embodiment

[0046] Example 3 The removal effect of Lactobacillus fermentum KDB-8 on cholesterol

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

[0048] After activation of Lactobacillus fermentum KDB-8 strain, it was inoculated into MRS medium and cultured overnight at 37°C as seed liquid.

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

[0050] Then, the seed solution of Lactobacillus fermentum KDB-8 was inoculated into the above-mentioned MRS medium, and after culturing at 37°C for 24 hours, the final cholesterol content in the medium was measured again to calcula...

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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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