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Extraction method of bile salt hydrolase in Lactococcus lactis and Streptococcus thermophilus

A technology of bile salt hydrolysis enzyme and Lactococcus lactis, which is applied in the field of extraction of bile salt hydrolysis enzymes from Lactococcus lactis and Streptococcus thermophilus, to achieve the effects of simple extraction process, short fermentation cycle and convenient source of raw materials

Active Publication Date: 2010-11-24
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although there is also a patent report on the research on the effect of Lactobacillus casei on lowering serum cholesterol abroad: Lactobacillus casei bd-II stain and used to reduce blood cholesterol (20060127380), however, there are almost no reports on Lactococcus lactis subsp. Patent report on the production of bile salt hydrolase and exopolysaccharide by Streptococcus and other strains
Although there are "a kind of Lactobacillus casei bile salt hydrolysis enzyme and its production method and special production strain (200610112696.2)", "a kind of bifidobacterium exopolysaccharide and its production method and special production strain (200510105465.4)", "cheese Lactobacillus exopolysaccharide and its crude product, preparation method and application (200510028970.3)" three patent reports, but there is no domestic patent on the extraction method of bile salt hydrolysis enzyme and exopolysaccharide in Lactococcus lactis subsp. lactic acid and Streptococcus thermophilus. Related article reports and patent reports

Method used

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  • Extraction method of bile salt hydrolase in Lactococcus lactis and Streptococcus thermophilus
  • Extraction method of bile salt hydrolase in Lactococcus lactis and Streptococcus thermophilus
  • Extraction method of bile salt hydrolase in Lactococcus lactis and Streptococcus thermophilus

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1. Screening and Identification of Lactococcus lactis subsp.lactis KS4 and Streptococcus thermophilus Tx with High Efficiency in Lowering Cholesterol and High Production of Bile Salt Hydrolase and Exopolysaccharide

[0035] 1. Screening of high-efficiency cholesterol-lowering and high-production bile salt hydrolase strains

[0036] Take 5mL of the filtrate of Zangling mushroom and pour it into 45mL sterile saline with glass beads, shake it fully for 30min to make a bacterial suspension, and then dilute it 10 times to 10 -4 ~10 -6 diluted bacterial solution. Inoculate the plate with the dilution pouring method, pour into the MRS selective medium containing calcium carbonate and natamycin, incubate at 37°C for 24 hours, pick a single colony with a dissolution circle and inoculate it in the MRS slant medium, and incubate at 37°C Cultivate for 24h. Gram staining, depending on the individual shape and purity. Inoculate in MRS liquid medium, enrich the bacteria fo...

Embodiment 2

[0062] Example 2, using Lactococcus lactis subspecies lactic acid KS4 and Streptococcus thermophilus Tx to extract bile salt hydrolyzing enzyme

[0063] 1. Fermentation process conditions for high production of bile salt hydrolase

[0064] Medium composition (w / v) for culturing Lactococcus lactis subsp. lactis KS4, Streptococcus thermophilus Tx high-yielding bile salt hydrolase: glucose 2%, soybean peptone 2%, MnSO 4 0.025%, beef extract 1%, yeast extract 0.5%, Tween 80 0.1%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, diammonium citrate 0.2%, magnesium sulfate 0.058%, distilled water 1000mL, 0.07MPa sterilization 20min.

[0065] Fermentation process conditions for high production of bile salt hydrolase: fermentation temperature 37° C., fermentation time 12 hours, initial pH value of medium 6.0, inoculum size 2%.

[0066] Under these fermentation conditions, the enzyme activity of Lactococcus lactis subsp. lactis KS4 was 11.84 times that before optimization.

...

Embodiment 3

[0072] Example 3, Utilizing Streptococcus thermophilus Tx and Lactococcus lactis subsp. lactic acid KS4 to extract exopolysaccharide

[0073] 1. Fermentation conditions for high-yield exopolysaccharide

[0074] Medium components (w / v) for cultivating Streptococcus thermophilus Tx and Lactococcus lactis subsp. lactis KS4 high-yielding exopolysaccharide: 6% sucrose, 3% tryptone, 0.025% manganese sulfate, 1.0% beef extract, yeast Powder 0.50%, Tween 80 0.10%, dipotassium hydrogen phosphate 0.20%, sodium acetate 0.50%, diammonium citrate 0.20%, magnesium sulfate 0.058%, distilled water 1000mL, 0.07MPa sterilization for 20min.

[0075] Fermentation conditions for high exopolysaccharide production: fermentation temperature 32° C., fermentation time 16 hours, initial pH value of medium 6.5, inoculum size 3%.

[0076] Under these fermentation conditions, the yield of Streptococcus thermophilus Tx exopolysaccharide was 2.6 times that before optimization.

[0077] 2. Extraction method...

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Abstract

The invention relates to an extraction method of bile salt hydrolase (BSH) and extracellular polysaccharide (EPS) in Lactococcus lactis and Streptococcus thermophilus, which is suitable for developing and producing novel functional foods by using BSH and EPS as functional factors and using EPS as a natural stabilizer in functional dairy products. In the invention, Lactococcus lactis subsp. lactis KS4 (CGMCC No.1807) and Streptococcus thermophilus Tx (CGMCC No.1810) of high-yield BSH and EPS are utilized, and the fermentation conditions and the extraction method are optimized to obtain crude products of BSH and EPS of lactic acid bacteria. A proper ultrasonic cell breakage extraction method is adopted to solve the problem that intracellular BSH can not be extracted easily; and the extraction conditions of the EPS are optimized to improve the extraction ratio of the EPS. The invention has simple fermentation process and extraction method for preparing BSH and EPS, shorter fermentation period, low requirements for equipment and lower cost, and is suitable for industrialized production.

Description

technical field [0001] The invention relates to a method for extracting bile salt hydrolase (Bile Salt Hydrolase, BSH) and exopolysaccharide (EPS) from Lactococcus lactis and Streptococcus thermophilus, and is suitable for BSH and EPS in functional dairy products as functional factors and EPS is used as a natural stabilizer to develop and produce new functional foods. Background technique [0002] Probiotics refer to a class of microorganisms that improve the microecological balance of specific parts of the host through intestinal colonization and have several other beneficial physiological functions. 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 can stimulate specific or non-specific immune responses, and have the effect of immune enhancement. The live bacteria of probiotic products should have stable metabolism ...

Claims

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Application Information

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IPC IPC(8): C12N9/14C12P19/04C12R1/01C12R1/46
Inventor 刘慧张红星钟德寿艾启俊董瑞婷
Owner BEIJING UNIV OF AGRI
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