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Preparation process of recombinant spirulina superoxide dismutase (SOD)

A superoxide and preparation technology, applied in the direction of oxidoreductase, recombinant DNA technology, botanical equipment and methods, etc., can solve the problems of organic solvent residue, low SOD content, low yield, etc.

Inactive Publication Date: 2016-02-17
鄂尔多斯市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The second is to extract SOD from plants, because the SOD content in plants is too low, it is difficult to achieve industrialization
This production process has some shortcomings such as organic solvent residue, complicated extraction procedure, high product cost, long production cycle, low yield, low purity, and poor product safety.

Method used

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  • Preparation process of recombinant spirulina superoxide dismutase (SOD)
  • Preparation process of recombinant spirulina superoxide dismutase (SOD)
  • Preparation process of recombinant spirulina superoxide dismutase (SOD)

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Embodiment Construction

[0104] The present invention will be further described in conjunction with accompanying drawing:

[0105] Such as figure 1 As shown, this study used Spirulina platensis platensis in Alkali Lake of Ordos Plateau as the experimental material, cloned and expressed its SOD coding gene, and established a large-scale recombinant SOD fermentation and purification process, on this basis, in-depth analysis of the recombinant expression product biological academic characteristics.

[0106]Ordos Plateau Alkali Lake Spirulina platensis was fished and purified from the sandy Alkali Lake in Ordos Plateau, cultivated with Zarrouk medium at room temperature, and was donated by Professor Qiao Chen from Inner Mongolia Agricultural University. Competent Escherichia coli DH5α is a product of Biomed Company, and competent Escherichia coli JM109 and BL21(DE3) are products of TaKaRa Company.

[0107] Construction and expression of prokaryotic expression vector of Spirulina sod gene

[0108] Extr...

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Abstract

The invention relates to a preparation process of recombinant spirulina superoxide dismutase (SOD) and belongs to the SOD preparation technology. The preparation process includes: cloning the encoding genes sod of spirulina platensis SOD, building a prokaryotic expression vector pET30a-sod, and expressing recombinant SOD in escherichia coli BL21(DE3); building a fermentation tank fermentation process of recombinant bacteria and a purification process of recombinant protein; measuring the protein concentration and specific activity of recombinant spirulina SOD; analyzing the natural molecular weight of the recombinant spirulina SOD through non-denaturing gel electrophoresis, and using a nitrotetrazolium blue chloride liquid dying method in combination with an inductively coupled plasma optical emission spectrometer to measure metal elements contained by pure enzyme so as to determine the type of the recombinant spirulina SOD; incubating pure enzyme liquid through heat treatment of different temperatures and solutions of different pH, and determining the stability of the pure enzyme. The preparation process has the advantages that the production process of the recombinant spirulina SOD is built, and the defects of residual organic solvent, complex extraction procedure, high product cost, long production cycle, low yield, low purity and poor product safety when the physicochemical technology is used for extracting the SOD are overcome.

Description

technical field [0001] The invention relates to a method for preparing superoxide dismutase, in particular to a method for preparing superoxide dismutase of spirulina platensis by using gene recombination technology. Background technique [0002] Superoxide dismutase (Superoxide Dismutase, SOD) is an active substance derived from living organisms and an anti-aging metalloenzyme. Its essence is protein. It was first discovered and extracted from bovine red blood cells by American scientists McCord and Fridovich in 1969. . In the normal metabolic process, the body will produce free radicals, which will be toxic to cells if they cannot be removed in time, and SOD can catalyze the disproportionation reaction of superoxide anion free radicals, converting superoxide free radicals in cells into oxygen and Hydrogen peroxide, so as to achieve the purpose of balancing oxygen free radicals in the body, can be divided into Fe-SOD, Mn-SOD and Cu / Zn-SOD according to the different metal i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/70C12R1/645
Inventor 高金亮王文杰姜晓杰
Owner 鄂尔多斯市疾病预防控制中心
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