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Low-temperature catalase and its preparation method and application

A catalase and low temperature technology, which is applied to low temperature catalase and its preparation, the preparation and application of low temperature catalase, the application field of low temperature catalase in removing H2O2, and can solve the problem of poor enzyme activity , restricting application, poor thermal stability and other problems, to achieve the effect of high enzyme activity and excellent thermal stability

Active Publication Date: 2021-03-19
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing catalase has defects such as poor thermal stability or poor enzyme activity at low temperature to varying degrees, which seriously restricts its application in production practice and needs to be improved urgently

Method used

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  • Low-temperature catalase and its preparation method and application
  • Low-temperature catalase and its preparation method and application
  • Low-temperature catalase and its preparation method and application

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0044] Example 1 Isolation, induced expression and purification of catalase Cat10171

[0045] 1 Experimental method

[0046] 1.1 Isolation of catalase Cat10171

[0047] Isolation of catalase gene from Tianshan soil macrogene cat10171 , and synthesized by GenScript containing the catalase gene cat10171 The recombinant plasmid pET28a- cat10171 .

[0048] 1.2 Induced expression of catalase Cat10171 in Escherichia coli

[0049] The recombinant plasmid pET28a- cat10171 Transformation into E. coli by heat shock method E . coli In BL21(DE3), spread on LB plates containing kanamycin Kana, culture overnight at 37 °C; pick 3 single clones, inoculate into 3 mL of LB liquid medium containing 100 μg / mL Kana, at 37 °C , culture overnight at 200 rpm; use the universal primer T7 / T7ter on pET28a to perform PCR verification on the cultured single clones; store the positive single clones in glycerol tubes.

[0050] Pick positive single clones into 3 mL of LB liquid medium containi...

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Abstract

The invention discloses low-temperature catalase, its preparation method and application. The present invention separates the catalase gene from the Tianshan soil macrogene, and its polynucleotide sequence is shown in SEQ ID No.1, encoding 502 amino acids and a stop codon. The optimal temperature of the catalase of the present invention is 10-20°C, and it has high enzyme activity in the low temperature range of 0-30°C, has the characteristics of a low-temperature enzyme, and can play a role in removing H in a low-temperature environment. 2 o 2 role. The catalase of the present invention has good thermal stability, and has good thermal stability at 40°C. After 3 h of treatment, 74% of the enzyme activity still remains, and has certain thermal stability at 50°C. After 0.5 h of treatment 78% of the enzyme activity still remained. The optimal pH of the catalase of the present invention is 8.0. The present invention further provides the catalase as an antioxidant in removing H 2 o 2 in the application.

Description

technical field [0001] The present invention relates to catalase, particularly relate to low-temperature catalase and preparation method thereof, the present invention further relates to this low-temperature catalase in scavenging H 2 o 2 The application in the present invention belongs to the field of preparation and application of low-temperature catalase. Background technique [0002] Catalase (Catalase, CAT, EC1.11.1.6) is a kind of specific catalytic H 2 o 2 decomposed into H 2 O and O 2 Enzymes widely exist in various bacteria, fungi, plants and animals. It is a class of antioxidants that can effectively remove H in cells 2 o 2 , to avoid H 2 o 2 Harmful effects on cells occupy a very important position in the defense system of organisms. In addition, CAT can be used to remove excess H in industrial production 2 o 2 , is an important industrial enzyme preparation widely used in medical, food and textile industries. Low-temperature CAT has high catalytic ef...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08C12N15/53C12N15/70C12N1/21A23C7/04C12R1/19
CPCA23C7/04C12N9/0065C12N15/70C12Y111/01006
Inventor 刘晓青伍宁丰关菲菲田健许国顺
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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