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Low temperature beta-galactosidase strain, low temperature bata-galactosidase and its production process

A technology of galactosidase and production process, which is applied in the field of low-temperature β-galactosidase and its production process, and can solve problems such as difficulty in meeting market and production needs, low enzyme activity, etc.

Inactive Publication Date: 2007-03-21
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-activity low-temperature β-galactosidase is required for fresh milk cold storage and fresh-keeping processing, and the production of low-lactose milk and low-lactose dairy products. market and production needs

Method used

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  • Low temperature beta-galactosidase strain, low temperature bata-galactosidase and its production process
  • Low temperature beta-galactosidase strain, low temperature bata-galactosidase and its production process
  • Low temperature beta-galactosidase strain, low temperature bata-galactosidase and its production process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Preservation and rejuvenation of bacterial strain L2004

[0038] A strain of low-temperature lactase-producing bacteria L2004 was isolated from the cold storage of Urumqi Meat Union Factory. It was identified as Bacillus brevius. The optimal reaction temperature of the low-temperature lactase produced was 33°C, and the half-life of the enzyme was about 50 minutes at 40°C.

[0039] Two methods were used for strain preservation, that is, liquid paraffin preservation method and vacuum freeze-drying method. The former is used for short-term preservation of strains (3-6 months), and the latter is used for long-term preservation of strains produced (6 months-several years).

[0040] Activation of preserved strains: take the preserved strains, use an inoculation needle to pick out the strains and inoculate them into a test tube filled with 2 ml of sterile water, and shake for 10 minutes. Then use a sterile pipette to take 0.2 ml and add it to the slant medium TS...

Embodiment 2

[0043] Example 2: Detection method for low temperature lactase activity of bacterial strain L2004

[0044] Use ONPG as the substrate for determination, take 1ml of enzyme solution in a test tube and preheat it in a 30°C water bath for 10 minutes, add 0.5mL pH6.5 0.1mol / L sodium phosphate buffer solution (1mol / L NaH 2 PO 4 70mL with 1mol / L Na 2 HPO 4 30mL buffer solution was mixed and diluted 10 times) and 0.2mL ONPG solution (pH6.5. 0.1mol / L sodium phosphate preparation) was reacted for 5min, and 0.8ml 5% Na 2 CO 3 Solution to stop the enzyme reaction, measure OD 420nm value. Under the above conditions, the amount of enzyme that hydrolyzes ONPG to generate 1 μmol o-nitrophenol per minute is 1 ONPG enzyme activity unit.

[0045] 1. Definition

[0046] 1 gram of solid enzyme powder (or 1ml of liquid enzyme), under certain temperature and pH conditions, hydrolyzes ONPG (o-nitrophenol galactoside) for 1 minute to produce 1umol tyrosine as an enzyme activity unit, expresse...

Embodiment 3

[0084] Embodiment 3: the measurement of the temperature characteristics of the bacterial strain of the present invention and its enzyme production

[0085] Medium: tryptone 15g, soybean peptone 5g, sodium chloride 5g, lactose 2.5g, dipotassium hydrogen phosphate 2.5g, water 1000ml, pH6.5.

[0086] At 4°C, 20°C, 42°C, 200 rpm, 60ml material / bottle (500ml shake flask), add cotton plug, culture time 6d, 2d, 1d respectively, take 5ml culture suspension in 50ml centrifuge tube , using a sonicator (broken wall) in an ice-water bath. Break the wall for 5 minutes at an intensity of 10 to release the enzyme from the cells. Take it out with a spectrophotometer at a wavelength of 420nm and a 10mm cuvette, and use a tube that does not meet ONP as a blank, and measure its absorbance respectively. At the same time, use a spectrophotometer at a wavelength of 620nm and a 10mm cuvette to measure the absorbance of the tube without inoculated culture medium as a blank.

[0087] The results sh...

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Abstract

The present invention is low temperature brevibacillus for producing beta-galactosidase and the production process of beta-galactosidase with the brevibacillus strain. By means of preservation, rejuvenation and breeding of the brevibacillus strain (CGMCC. No.1424) and optimized fermentation process, the present invention has short fermentation period and stable fermentation process to obtain beta-galactosidase with excellent high temperature activity and high enzyme activity. The product may be used widely in fermented milk product, beverage, food, medicine and other fields.

Description

field of invention [0001] The invention relates to the field of microorganisms and microbial fermentation. Specifically, the present invention relates to a Brevibacillus strain producing low-temperature β-galactosidase, low-temperature β-galactosidase and a production process thereof. Background technique [0002] β-Galactosidase (EC.3.2.1. and Glucose 23), also known as lactase, can not only hydrolyze lactose into galactose and glucose that are easy to absorb and have good sweetness, but also A transglycoside reaction will occur to generate galacto-oligosaccharides with various physiological functions. This sugar is a bifidobacterium value-added factor, which is difficult to be digested by the human body. It can be used to produce low-lactose foods, improve the flavor and nutrition of dairy products, and relieve various adverse effects such as diarrhea and abdominal distension caused by lactose intolerance patients who eat dairy products. reaction. It can also improve co...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/38C12R1/08
Inventor 史应武娄恺魏东欧提库尔常玮
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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