超氧化物歧化酶重组酵母的构建及其发酵条件的优化

Construction and Expression of Recombinant Superoxide Dismutase in Pichia pastoris

DOI：10.3969/j.issn.1673-1689.2017.09.004

中文关键词: 人源铜锌超氧化物歧化酶 基因工程 巴斯德毕赤酵母 表达 优化

英文关键词: superoxide dimutase,gene engineering,Pichia pastoris,expression optimization

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中文摘要:

超氧化物歧化酶（SOD）由于具有高效清除氧自由基的作用而广泛用于食品、日化和医药领域。作者研究了提高SOD产量的基因工程新方法。利用密码子优化后人源铜锌超氧化物歧化酶基因，构建亚克隆及表达载体，最终转化进巴斯德毕赤酵母进行表达。通过正交试验，分析工程菌的最适摇瓶表达条件。摇瓶正交试验确定最优表达条件为：28 ℃，诱导剂体积分数1.0%，接种体积分数1.0%，最优表达水平为2 339.4 U/mL，与现有的一些研究相比，其表达量提高了近3倍。这些结果表明：对人源铜锌SOD经过适当的密码子优化和发酵条件优化，使酵母表达量显著提高，有助于解决SOD原料来源的局限。

英文摘要:

Superoxide dismutase （SOD） can be widely used in the field of food，cosmetic and medicine for its high capacity to scavenge free radicals. This paper reported a method of gene engineering attempting to solve the issue of SOD production. The gene of Cu，Zn-SOD with the yeast bias codon was firstly chemically synthesized and integrated into the genome of Pichia pastoris and the expression condition was optimized through orthogonal experiment. It was found that the maximum expression level of SOD was nearly three times greater than some reserach reported and the optimal temperature，methanol concentration and inoculation were 28 ℃，1.0% and 1.0% respectively. These results suggest that the problem of SOD production can be solved by improving the expression level of P.pastoris through the optimization of gene codon and fermentation conditions.

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