Optimization of the purification methods for recovery of recombinant growth hormone from Paralichthys olivaceus
文献类型: 外文期刊
第一作者: Liu Bin
作者: Liu Bin;Zang Xiaonan;Zhang Xuecheng;Mu Xiaosheng
作者机构: Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China;Ocean Univ China, Coll Marine Life Sci, Qingdao 266003, Peoples R China
关键词: Paralichthys olivaceus;growth hormone;fusion expression;purification;receptor binding activity
期刊名称:JOURNAL OF OCEAN UNIVERSITY OF CHINA ( 2020影响因子:0.913; 五年影响因子:1.012 )
ISSN: 1672-5182
年卷期: 2013 年 12 卷 1 期
收录情况: SCI
摘要: This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone (r-fGH) was expressed by Escherichia coli in form of inclusion body or as soluble protein under different inducing conditions. The inclusion body was renatured using two recovery methods, i.e., dilution and dialysis. Thereafter, the refolded protein was purified by Glutathione Sepharase 4B affinity chromatography and r-fGH was obtained by cleavage of thrombin. For soluble products, r-fGH was directly purified from the lysates by Glutathione Sepharase 4B affinity chromatography. ELISA-receptor assay demonstrated that despite its low receptor binding activity, the r-fGH purified from refolded inclusion body had a higher yield (2.605 mg L-1) than that from soluble protein (1.964 mg L-1). Of the tested recovery methods, addition of renaturing buffer (pH 8.5) into denatured inclusion body yielded the best recovery rate (17.9%). This work provided an optimized purification method for high recovery of r-fGH, thus contributing to the application of r-fGH to aquaculture.
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