Beta-galactosidase strain, beta-galactosidase and its production process

A technology of galactosidase and production process, which is applied in the field of microorganisms and microbial fermentation, and can solve the problems of fresh milk deterioration, high cost, and long time occupied by equipment

Inactive Publication Date: 2009-01-07
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

Because the chemical reaction speed is slow at room temperature and takes a long time, the fresh milk is easily contaminated by bacteria and deteriorates, and the production process is prolonged and the equipment takes a long time
In addition, the amount of enzyme to be added to shorten the hydrolysis time is large, so the cost is also high

Method used

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  • Beta-galactosidase strain, beta-galactosidase and its production process
  • Beta-galactosidase strain, beta-galactosidase and its production process
  • Beta-galactosidase strain, beta-galactosidase and its production process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the present invention selects the preservation and rejuvenation of bacterial species

[0040] A lactase-producing strain G2005 was isolated in Urumqi cold storage, which was identified as Lactococcus. The optimal reaction temperature of the lactase produced was 50℃, and the half-life of the enzyme was about 50 minutes at 60℃.

[0041] Two methods were used for the preservation of strains, namely liquid paraffin preservation and vacuum freeze-drying. The former is used for short-term preservation (4-6 months) of strains, and the latter is used for long-term preservation (6 months-several years) of production strains.

[0042] Activation of the preserved strains: Take the preserved strains, pick the strains with an inoculating needle 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 plate medium TSA (tryptone 15g, soy peptone 5g, dipotassium hydrogen p...

Embodiment 2

[0045] Embodiment 2: G2005 lactase activity detection method

[0046] According to FCC standards, fourth edition, July 1, 1996, pages 801 to 802 / Lactase (neutral) β-gulactosidase / activity. Lactococcus lactase test method and G2005 lactase characteristics, the temperature was set at 50°C and the pH was 7.0 during the test.

[0047] 1. Definition

[0048] 1 gram of solid enzyme powder (or 1 ml of liquid enzyme), under certain temperature and pH conditions, hydrolyze ONPG (o-nitrophenol galactoside) in 1 minute to produce 1 umol of tyrosine as an enzyme activity unit, with u / g ( u / ml) indicated.

[0049] 2. Colorimetry

[0050] 2.1 Principle

[0051] G2005 lactase hydrolyzed ONPG substrate at 50℃ and pH 7.0 to produce ONP containing phenolic group. Under alkaline conditions, ONPG turned yellow, and its enzymatic activity was calculated by spectrophotometer.

[0052] 2.2 Reagents and solutions

[0053] 2.2.1 Preparation of sodium phosphate buffer

[0054] Take 1mol / LNa resp...

Embodiment 3

[0089] Example 3: Enzymatic properties of strains of the present invention at different temperatures

[0090] Medium: tryptone 15g, soy peptone 5g, sodium chloride 5g, lactose 2.5g, dipotassium hydrogen phosphate 2.5g, water 1000ml, pH7.

[0091] At 30°C, 200 rpm, 60ml of material / bottle (500ml shaker), add cotton plugs, culture for 48h, take 5ml of cultured bacterial suspension into a 50ml centrifuge tube, and use an ultrasonicator (breaking the wall) in an ice-water bath. The wall was broken at 10 strength for 5 min to release the enzyme from the cells. Take out with a spectrophotometer at a wavelength of 420nm, a 10mm cuvette, take the tube without ONP as a blank, and store at 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C , 50 ℃, 60 ℃ were measured the absorbance, the calculated enzyme activities were 5.53, 5.86, 6.06, 6.19, 6.46, 7.85, 7.85, 9.83, 10.76, 13.20, 7.78. Therefore, the Lactococcus sp. strain of the present invention has the characteristics that temperat...

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Abstract

The present invention is lactococcus for producing beta-galactosidase and the production process of beta-galactosidase with the lactococcus strain. By means of preservation, rejuvenation and breeding of the lactococcus strain (CGMCC. No.1425) 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 lactococcus strain producing a β-galactosidase, a β-galactosidase and a production process thereof. Background technique [0002] β-galactosidase (EC3.2.1.23), also known as lactase, widely exists in various animals, plants and microorganisms. The initial application of this enzyme was to reduce the lactose content of dairy products by utilizing its lactose-hydrolyzing properties. With the development of biotechnology, β-galactosidase with certain excellent characteristics and higher enzyme activity can be obtained, and the gene structure encoding this enzyme has been well understood, and its reaction mechanism has been explored, so that β-galactosidase Glycosidases are not only used more and more widely in the food industry, but also play an important role in the fields of biotechnology such as genetic engineering, enz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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