Recombinant superoxide dismutase as well as coding gene and preparation method thereof

A technology of superoxide and dismutase, applied in botany equipment and methods, biochemical equipment and methods, oxidoreductase, etc., can solve the problems of restricting the development and application of SOD, difficult application, etc., and achieve good resistance to commonly used preservatives The effect of good heat resistance, good heat stability and good acid and alkali resistance

Inactive Publication Date: 2021-04-30
ZHEJIANG UNIV OF TECH +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although SOD has been proven to have a very good effect, it is still difficult to have practical application at present, mainly because SOD has certain defects in high temperature resistance, acid and alkali resistance, and corrosion resistance, which greatly limits the use of SOD in Developments and applications in the fields of medicine, cosmetics and food

Method used

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  • Recombinant superoxide dismutase as well as coding gene and preparation method thereof
  • Recombinant superoxide dismutase as well as coding gene and preparation method thereof
  • Recombinant superoxide dismutase as well as coding gene and preparation method thereof

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preparation example Construction

[0027] The preparation of embodiment recombinant superoxide dismutase

[0028] 1. High-yielding mutant high-temperature resistant SOD strain was obtained

[0029] Using the gene encoding SOD in thermophilic bacteria HB27 (purchased from ATCC cell bank in the United States) as a template, PCR amplification was performed with the following primer sequences to obtain the target gene:

[0030] Forward primer: 5'-AAGAATTCATGCCGTACCCGTTCAAGCT-3'(SEQ ID NO.1)

[0031] Reverse primer: 5'-CTGTCGACTCAGGCTTTGTTGAAGAAC-3' (SEQ ID NO.2);

[0032] PCR system:

[0033]

[0034] PCR reaction program: 98°C 10min pre-denaturation; 98°C 30sec high-temperature denaturation, 55°C 30sec annealing, 72°C 1min extension, cycle 30 times; finally 72°C 1min complete extension and 10°C incubation to fully amplify the reaction;

[0035] The amplified product was recovered with a recovery kit (purchased from Sangon Bioengineering Shanghai (Co., Ltd.)), double-digested with enzymes (EcoRI and SalI), an...

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Abstract

The invention belongs to the field of organic matter separation and purification, and particularly relates to recombinant superoxide dismutase (recombinant SOD for short), a gene for encoding the recombinant SOD, a related expression vector, a transgenic cell line and host bacteria, and a preparation method of the recombinant SOD. The amino acid sequence of the recombinant SOD is shown as SEQ ID NO.4 in a sequence table. The gene sequence of the gene for encoding the recombinant SOD is shown as SEQ ID NO.3 in a sequence table. Tests prove that the recombinant SOD disclosed by the invention has excellent thermal stability, excellent acid-base tolerance and relatively good common preservative tolerance, and can be widely applied to the fields of medicines, cosmetics and foods.

Description

technical field [0001] The invention belongs to the field of separation and purification of organic matter, in particular to a recombinant superoxide dismutase (superoxide dismutase, SOD) and its coding gene and preparation method. Background technique [0002] Free radicals have become a research hotspot in life sciences since Moses Gomberg discovered that they are closely related to life activities in 1900. In 1956, Denham Harman introduced the concept of free radicals into life science based on the phenomenon of increased free radicals in radiation damage, and proposed the hypothesis of free radical aging. In the course of normal life, free radicals in the body are constantly produced to maintain life activities, but they are also constantly removed to maintain the normal level of free radicals. Excessive free radicals can lead to various diseases, such as tumors, atherosclerosis, myocardial infarction, diabetes, cataracts, inflammation, hypoxic or ischemic damage, strok...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/70C12N1/21
CPCC12N9/0089C12N15/70C12Y115/01001
Inventor 章志豪李海龙孟凡国储消和
Owner ZHEJIANG UNIV OF TECH
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