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Glycosyl transferred beta-galactosidase bacteria screening method

A technology of galactosidase and transglycosidation, which is applied in the field of screening microorganisms and can solve the problems of not screening transglycosidase β-galactosidase

Inactive Publication Date: 2007-05-02
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the search for microorganisms that can produce strong transglycosyl active β-galactosidase has become a research hotspot at present. However, in the relevant literature retrieved, there is currently no screening for the production of transglycosyl β-galactosidase in the world. report on bacterial methods

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Screening transglycosyl β-galactosidase producing bacteria from soil

[0034] 1. Isolation and purification of growth strains that can use lactose as the only carbon source

[0035] Add 10g of soil sample to 100mL of culture solution with lactose as the only carbon source for enrichment culture, and dilute the culture solution with sterile water to 10 -1 、10 -3 、10 -5 、10 -7 For each gradient, 200 μL was evenly spread on the lactose selection plate. Bacterial screening plates were incubated at 37°C for 24 hours, and fungal screening plates were incubated at 28°C for 48-60 hours. 10 -5 There are many types of colonies grown on gradient plates, and they are easy to purify. Colonies with different shapes were picked, separated and purified by streaking on the plate, and repeated 3 times. Perform microscopic examination on the isolated strains to determine the purity: the bacteria use Staphylococcus aureus (Staphylococcusaureus) and Escherichia coli (Esche...

Embodiment 2

[0054] Example 2: Screening transglycosyl β-galactosidase producing bacteria from milk dregs

[0055] 1. Isolation and purification of growth strains that can use lactose as the only carbon source

[0056] Add 10g of milk dregs to 100mL culture medium with lactose as the sole carbon source for enrichment culture, and dilute the culture medium with sterile water to 10 -2 、10 -4 、10 -6 、10 -8 For each gradient, 200 μL was evenly spread on the lactose selection plate. Bacterial selection plates were incubated at 37°C for 24 hours, and fungal selection plates were incubated at 28°C for 60 hours. 10 -6 There are many types of colonies grown on gradient plates, and they are easy to purify. Colonies with different shapes were picked, separated and purified by streaking on the plate, and repeated 3 times. Perform microscopic examination on the isolated strains to determine the purity: the bacteria use Staphylococcus aureus (Staphylococcus cmreus) and Escherichia coli (Escherich...

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Abstract

The invention disclosed a screen selecting method for the glycosyl-transferase beta-galactosidase producing bacterium, comprising the following steps: using the lactin as the only scource of carbon, after enriching the microbios, separating it with flat, then screening the beta-galactosidase hydrolyzation activity and galactosyl activity and getting the glycosyl-transferase beta-galactosidase producing bacterium. With utilizing the method of the invention, it is easy and effective to obtain the glycosyl-transferase beta-galactosidase producing bacterium from the nature, and also obtain the high active glycosyl-transferase beta-galactosidase producing bacterium. The method provides new source of bacterial for exploiting the microbiological industry, especially for producting oligo-galactose.

Description

technical field [0001] The invention relates to a method for screening microorganisms, in particular to a method for screening transglycosyl beta-galactosidase-producing bacteria. technical background [0002] Galactooligosaccharide is one of the natural components in breast milk, and its most important and fundamental physiological function comes from its effect on the proliferation of bifidobacteria. Because galactooligosaccharide has good heat stability and acid resistance, its health care effect has aroused great concern of people. At present, the production of galacto-oligosaccharides is mainly synthesized by microbial enzymatic methods, that is, β-galactosidase or lactase uses lactose as a substrate to transglycosylate synthesis. Since the synthesis reaction catalyzed by glycosidase is a reversible reaction, that is, the transglycosylated product can be hydrolyzed by the enzyme as a substrate, the synthesis efficiency is not very high. Therefore, the search for micro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12N9/38
Inventor 肖敏卢丽丽
Owner SHANDONG UNIV