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Heat-resistant and stress-resistant sod and its coding gene and application through genetic engineering transformation

A technology encoding gene and amino acid is applied in the field of thermostable superoxide dismutase and its encoding gene and in the field of thermostable oxide dismutase, which can solve the problem that denaturants, organic solvents are easily inactivated, and the product transportation and sales cycle is shortened. , limiting product processing technology and other issues, to achieve the effects of good adaptability, good stress resistance, and simple recombinant expression methods

Inactive Publication Date: 2018-11-02
北京迈恩泰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to the nutritional and health functions of SOD, many SOD products have appeared in the domestic market, such as beer, fruit, rice, oral liquid, etc. containing SOD. There are unfavorable factors such as short half-life, easy inactivation in case of high temperature, denaturants, and organic solvents
This not only makes the development of SOD-containing products more difficult, limits the processing technology of products, but also greatly shortens the validity period of SOD-containing products, forcing the storage, transportation and sales cycles of products to be shortened, and increasing operational difficulties.

Method used

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  • Heat-resistant and stress-resistant sod and its coding gene and application through genetic engineering transformation
  • Heat-resistant and stress-resistant sod and its coding gene and application through genetic engineering transformation
  • Heat-resistant and stress-resistant sod and its coding gene and application through genetic engineering transformation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Build code Sulfolobussolfataricus Fe-SOD full sequence gene ( Sssod ), and construct the N-terminal sequence encoding the recombinant SOD (SOD-GTNG_2215 and Sulfolobussolfataricus The recombination of Fe-SOD is combined into a recombination SOD) full sequence gene ( rSssod ) Clone. And measure the enzyme activity and stress resistance of the expressed protein.

[0042] 1. Extraction of total DNA from Bacillus thermophilus NG80-2 (CGMCC No. 1228)

[0043] In this example, the Bacillus thermophilus NG80-2 (Bacillus thermophilus) isolated from the formation water of the oil well in the Guan 69-8 block of the Dagang Oilfield in Tianjin, China ( Geobacillusthermodenitrificans This strain has been deposited in the General Microbiology Center of China Microbial Culture Collection, and its preservation number is CGMCC No. 1228). Take 3 mL of the fresh culture cultured overnight and collect the bacteria by centrifugation. The bacteria are suspended in 250 μL of 50 mM Tris buffer ...

Embodiment 2

[0076] Because of the special effects of SOD, it has a wide range of applications in the fields of medicine, daily chemical, food, agriculture and environmental protection. At present, the clinical application of SOD is mainly focused on anti-inflammatory (mainly patients with rheumatoid and inflammation caused by radiotherapy), in addition to certain autoimmune diseases (such as lupus erythematosus, dermatomyositis), emphysema, anti-inflammatory Cancer and oxygen poisoning have certain curative effects; in the food industry, they are mainly used as food additives and important functional base materials; in the cosmetics industry, they are mainly used as important functional ingredients for anti-inflammatory and anti-aging.

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Abstract

The invention discloses heat-resisting and stress-resisting SOD modified by adopting genetic engineering as well as an encoding gene and application thereof. The superoxide dismutase has protein with the following amino acid residue sequence: SEQ ID NO.1. An expression method comprises the following steps of constructing a heat-resisting superoxide dismutase recombinant expression vector, introducing the constructed recombinant expression vector into a host cell, and inducing to express heat-resisting superoxide dismutase genes. The expression product has prominent heat resistance and good stress resistance, has resistance on acid and alkali, a cleaning agent, a protein denaturing agent and an inhibitor, is easy to purity and has good stability, and has a wide industrial application prospect.

Description

Technical field [0001] The invention relates to an enzyme and its encoding gene and application, in particular to a thermostable superoxide dismutase and its encoding gene and its use in thermostable oxide dismutase. Background technique [0002] Superoxide dismutase (SOD) is an enzyme containing metal ions discovered by American scientists Mccord and Fridovich in 1969 from bovine red blood cells. This enzyme can specifically remove superoxide anion free radicals generated in the biological oxidation process, and is the body's first line of defense against free radicals. It plays an extremely important role in maintaining the dynamic balance of free radical production and removal in biological organisms. effect. Based on the difference of combined metal prosthetic group, SOD can be divided into four categories: Cu / Zn-SOD, Mn-SOD, Fe-SOD and Ni-SOD. SOD has the effects of anti-inflammatory, anti-radiation, anti-tumor, anti-aging, etc., and is used in the clinic of viral diseases...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N1/15C12N1/19C12N1/21C12N15/11
CPCC12N9/0089C12Y115/01001
Inventor 王威李洺畅
Owner 北京迈恩泰生物科技有限公司
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