梅花鹿（Cervus nippon）Cu/Zn 超氧化物歧化酶在毕赤酵母中的高水平表达及其特性。
High-level expression of a sika deer (Cervus nippon) Cu/Zn superoxide dismutase in Pichia pastoris and its characterization.
机构信息
College of Biological Science and Technology, Fuzhou University, PR China.
出版信息
Environ Toxicol Pharmacol. 2013 Mar;35(2):185-92. doi: 10.1016/j.etap.2012.11.013. Epub 2012 Dec 3.
Production of a sika deer Cu/Zn-SOD was achieved in Pichia pastoris after the reconstituted expression vector pPIC9K was transformed into the strain GS115. By employing Saccharomyces cerevisiae secretion signal peptide (α-factor) under the regulation of the methanol-inducible promoter of the gene of alcohol oxidase 1 (AOX1), sika deer Cu/Zn-SOD with a molecular mass of 16kDa was expressed while recombinant sika deer Cu/Zn-SOD with an activity of 3500U/mL was obtained from a 5L bioreactor. After two successive steps of chromatography on DEAE-650C and Superdex75, recombinant sika deer Cu/Zn-SOD was obtained with 13.8% yield, 14.5-fold purification, and a specific activity of 3447U/mg. Its optimum temperature and optimum pH were 40°C and 7.0, respectively.
重组表达载体 pPIC9K 转化毕赤酵母 GS115 后，在甲醇诱导型醇氧化酶 1（AOX1）基因启动子的调控下，利用酿酒酵母分泌信号肽（α-因子），成功生产出分子量为 16kDa 的梅花鹿 Cu/Zn-SOD。从 5L 生物反应器中获得了活性为 3500U/mL 的重组梅花鹿 Cu/Zn-SOD。经过 DEAE-650C 和 Superdex75 两步连续层析，获得了 13.8%收率、14.5 倍纯化和 3447U/mg 的比活性的重组梅花鹿 Cu/Zn-SOD。其最适温度和最适 pH 值分别为 40°C 和 7.0。
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