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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, protracted production process, and slow chemical reaction speed

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

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

[0038] Embodiment 1: the present invention selects the preservation and rejuvenation of bacterial classification

[0039] A strain of lactase-producing bacteria G2005 was isolated from a cold storage in Urumqi, and was identified as Lactococcus. The optimal reaction temperature of the lactase produced was 50°C, and the half-life of the enzyme at 60°C was about 50 minutes.

[0040] 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 (4-6 months), and the latter is used for long-term preservation of strains produced (6 months to several years).

[0041] 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 plate medium TSA (tryptone 15g, s...

Embodiment 2

[0044] Embodiment 2: G2005 lactase activity detection method

[0045] According to FCC standard, fourth edition, July 1, 1996, page 801 to 802 / Lacttase (neutral) / (β-galactosidase) activity (Lacttase (neutral) β-gulactosidase / activity). Lactococcus lactase test method and G2005 lactase characteristics, during the test, the temperature is set at 50°C and the pH is 7.0.

[0046] 1. Definition

[0047] 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, expressed in u / g ( u / ml) said.

[0048] 2. Colorimetry

[0049] 2.1 Principle

[0050] G2005 lactase hydrolyzes ONPG substrate at 50°C and pH 7.0 to produce ONP containing phenolic groups. Under alkaline conditions, ONPG turns yellow, and its enzyme activity is calculated by measuring it with a spectrophotometer.

[0051] 2.2 Reagents and solutions

[0052] 2.2.1 Prep...

Embodiment 3

[0086] Embodiment 3: the enzymatic characteristic of strain of the present invention different temperatures

[0087] Medium: tryptone 15g, soy peptone 5g, sodium chloride 5g, lactose 2.5g, dipotassium hydrogen phosphate 2.5g, water 1000ml, pH7.

[0088] At 30°C, 200 rpm, 60ml material / bottle (500ml shake flask), add a cotton plug, incubate for 48h, take 5ml culture suspension in a 50ml centrifuge tube, and use an ultrasonic instrument (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 out with a spectrophotometer at a wavelength of 420nm, a 10mm cuvette, and a tube without ONP as a blank, at 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C , 50°C, and 60°C were measured for their absorbance, and 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, and 7.78, respectively. Therefore, the Lactococcus strain (Lacfococcus sp.) of the present invention ha...

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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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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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