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Lactic acid bacteria for producing bile salt hydrolase as well as screening method and application thereof

A technology for enzymolytic lactic acid bacteria and screening method, applied in the directions of microorganism-based methods, applications, biochemical equipment and methods, etc., can solve problems such as insufficient sensitivity, and achieve the effects of high sensitivity, small workload and simple operation

Active Publication Date: 2011-10-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, agar plate test is usually used for detection at home and abroad, but the sensitivity of this screening method is not enough, and it is only suitable for screening bacteria with high BSH activity (such as bifidobacteria).

Method used

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  • Lactic acid bacteria for producing bile salt hydrolase as well as screening method and application thereof
  • Lactic acid bacteria for producing bile salt hydrolase as well as screening method and application thereof
  • Lactic acid bacteria for producing bile salt hydrolase as well as screening method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 produces the qualitative screening method of bile salt hydrolase

[0026] 1. Starting strain

[0027] The starting strains are existing strains in the laboratory, including strains BBE1, BBE2, BBE3, BBE4, BBE5, BBE6, BBE7, BBE8, BBE10-211, BBE10-212, BBE10-215, BBE Kefir, Lactobacillus caseiZhang (LB.Z), Lactobacillus casei ATCC334, Lactobacillus acidophilus ATCC4356 and Lactobacillus rhamnosus ATCC7469, among which the first 12 strains were screened in this experiment, BBE10-212 screened from kimchi was identified and named as Lactobacillus casei BBE10-212, and has applied for a Chinese patent with the application number 201010238658.8 ; Lactobacillus casei Zhang (LB.Z) was screened from sour kumiss by Inner Mongolia Agricultural University; Lactobacillus caseiATCC334, Lactobacillus acidophilus ATCC4356 and Lactobacillus rhamnosus ATCC7469 were purchased from American Type Culture Collection (ATCC).

[0028] 2. Screening method

[0029] (1) Activation of...

Embodiment 2

[0040] Embodiment 2: the quantitative screening method of the lactic acid bacteria that produces bile salt hydrolase (ninhydrin method measures enzyme activity)

[0041] (1) Ninhydrin Chromogenic Solution

[0042] Dissolve ninhydrin at a ratio of 1% in 0.5M citric acid buffer (pH 5.5), take 0.5 mL of the above mixture and add 1.2 mL of glycerin and 0.2 mL of 0.5 M citric acid buffer (pH 5.5).

[0043] (2) Enzyme activity assay method

[0044] (1) Activation of bacteria

[0045] Lactic acid bacteria preserved in glycerol tubes at -70°C were inoculated with 1% inoculum into test tubes containing 2mL of MRS medium, and cultured at 37°C for 20 hours.

[0046] (2) shake flask culture

[0047] The above-mentioned activated bacterial solution was transferred to a 50mL Erlenmeyer flask containing 20mL MRS medium according to the inoculum amount of 1%, and cultured statically at 37°C until the stationary phase.

[0048] (3) Determination of enzyme activity

[0049] The above ferme...

Embodiment 3

[0066] Embodiment 3: the effect of lactic acid bacteria removing cholesterol in vitro

[0067] (1) Cholesterol-containing MRS liquid medium

[0068] First prepare a concentration of 10mg mL with absolute ethanol -1 After filtering and sterilizing the cholesterol solution with a microporous membrane with a pore size of 0.45 μm, it was added to sterilized MRS containing 0.30% sodium taurocholate, so that the final concentration of cholesterol was 100 μg mL -1 .

[0069] (2) Preparation of fermentation broth

[0070] The test bacteria were continuously passed for 3 generations in the MRS medium, and the 3rd generation culture was 5000r min -1 Centrifuge for 10 min, discard the supernatant, wash the bacteria with sterilized physiological saline for 3 times, adjust the concentration of the bacterial suspension to 3×10 9 CFU mL -1 . Inoculate into cholesterol-containing MRS liquid medium according to 2% inoculum amount, and incubate anaerobically at 37°C for 24h. At the same ...

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Abstract

The invention discloses lactic acid bacteria for producing bile salt hydrolase as well as a screening method and application thereof, belonging to the technical field of food biology. In the invention, an improved qualitative analysis and quantitative analysis combined method can be used for accurately screening a bile salt hydrolase bacterial strain in high efficiency; the method disclosed by the invention is used for screening a plant lactic acid bacterium strain with higher activity of bile salt hydrolase; and the bacterial strain and the bile salt hydrolase produced by the bacterial strain can be applied to the field of foods and can be used for reducing the content of cholesterol of human serum.

Description

technical field [0001] The invention relates to a lactic acid bacterium producing bile salt hydrolase, a screening method and application thereof, and belongs to the field of food biotechnology. Background technique [0002] Probiotics refer to microorganisms that are beneficial to human and animal organisms. It includes Lactobacillus casei, Streptococcus thermophilus, Bifidobacterium, Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus plantarum, Lactobacillus fermentum, etc. They can improve the ecological balance of intestinal microorganisms, control intestinal infection, reduce serum cholesterol levels, improve the utilization of lactose for people with lactose intolerance, and stimulate specific or non-specific immune responses, which have the effect of enhancing the body's immunity. It is mainly used in the production of fermented foods such as dairy products, meat products, aquatic products and some functional foods. [0003] High cholesterol content ...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/04C12N1/20A23L1/29C12R1/25A23L33/00
Inventor 陈坚董自星张娟堵国成李华钟
Owner JIANGNAN UNIV
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