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A heat-resistant and stress-resistant SOD modified by genetic engineering and its coding gene and application

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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  • A heat-resistant and stress-resistant SOD modified by genetic engineering and its coding gene and application
  • A heat-resistant and stress-resistant SOD modified by genetic engineering and its coding gene and application
  • A heat-resistant and stress-resistant SOD modified by genetic engineering and its coding gene and application

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Experimental program
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Effect test

Embodiment 1

[0035] build code Aeropyrumpernix The full sequence gene of Fe / Mn-SOD ( Apsod ) clone, and construct the N-terminal sequence encoding recombinant SOD (SOD-GTNG_2215 and Aeropyrumpernix The recombination of Fe / Mn-SOD combined into recombinant SOD) full sequence gene ( rApsod ) clones. And the enzyme activity and stress resistance of the expressed protein were measured.

[0036] 1. Extraction of total DNA from Bacillus thermodenitrophilus NG80-2 (CGMCC No. 1228)

[0037] In this example, the thermophilic denitrification bacillus NG80-2 ( Geobacillus thermodenitrificans The strain has been preserved in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures, and its preservation number is CGMCC No.1228). Take 3 mL of fresh culture cultured overnight, collect the bacteria by centrifugation, and suspend the bacteria in 250 μL of 50 mM Tris buffer (pH8 .0), add 10μL 0.4M EDTA (pH8.0), mix well and keep warm at 37°C for 20min, then ad...

Embodiment 2

[0071] Because SOD can specifically remove superoxide anion free radicals produced in the process of biological oxidation, it plays an extremely important role in maintaining the dynamic balance of free radical production and removal in biological organisms. It is widely used in medicine, daily chemical industry, food , agriculture and environmental protection and other fields have a wide range of application value. At present, the clinical application of SOD mainly focuses on anti-inflammation, anti-radiation, anti-tumor, anti-aging, etc., and is used for viral diseases, autoimmune diseases, myocardial ischemia and reperfusion syndrome, senile cataract, cardiovascular disease, Clinical treatment of radiation sickness and cancer; in the food industry, it can be added as an efficacy factor of health food; in cosmetics, it can be added for sun protection, anti-oxidation, skin aging prevention, and scar formation, etc.

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Abstract

The invention discloses high-temperature-resistant superoxide dismutase as well as an encoding gene and application thereof. The superoxide dismutase comprises protein with an amino acid residue sequence as shown in SEQ ID NO.1. The expression method comprises the following steps: establishing a recombinant expression vector containing a thermal-resistant superoxide dismutase gene, introducing the established recombinant expression vector into a host cell, and inducing so as to express the thermal-resistant superoxide dismutase gene. The expression product disclosed by the invention is outstanding in thermal resistance, good in adverse resistance, good in resistance to acid, alkali, organic solvents, detergent, protein denaturants and inhibitors, easy to purify, good in stability, and side in industrial application prospect.

Description

technical field [0001] The present invention relates to an enzyme and its coding gene and application, in particular to a heat-resistant superoxide dismutase and its coding gene and the heat-resistant superoxide dismutase. Background technique [0002] Superoxide dismutase (SOD for short) is an enzyme containing metal ions discovered by American scientists Mccord and Fridovich1969 from bovine red blood cells. This enzyme can specifically remove superoxide anion free radicals produced in the process of biological oxidation. It is the body's first line of defense against free radicals and plays an extremely important role in maintaining the dynamic balance of free radical production and removal in biological organisms. effect. Based on the different binding metal prosthetic groups, SOD can be divided into four categories: Cu / Zn-SOD, Mn-SOD, Fe-SOD, and Ni-SOD. SOD has anti-inflammatory, anti-radiation, anti-tumor, anti-aging effects, and is used in the clinical treatment of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/70C12R1/19
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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