Expression of Apostichopus japonicus lysozyme in the methylotrophic yeast Pichia pastoris

文献类型： 外文期刊

第一作者： Wang, Tingting

作者： Wang, Tingting;Xu, Yongping;Liu, Wenjia;Jin, Liji;Xu, Yongping;Jin, Liji;Sun, Yongxin;Wang, Tingting;Xu, Yongping;Liu, Wenjia;Sun, Yongxin;Jin, Liji

作者机构：

关键词： Antimicrobial activity;Apostichopus japonicus;Expression;Lysozyme;Pichia pastoris

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： Apostichopus japonicus (sea cucumber) is one of the economically important farmed echinoderm species in Northern China. As a crucial enzyme in innate immunity, lysozyme plays a key role in the overall defense against pathogens in A. japonicus. In the present study, a lysozyme gene from A. japonicus was cloned by PCR and expressed in Pichia pastoris using the expression vector pPIC9K. The expressed lysozyme had a molecular mass of ～14 kD, as shown by SDS-PAGE and Western-blotting. The expression condition was optimized, and the highest expression level was achieved by induction with 1% methanol at pH 5.0 for 120 h. The recombinant lysozyme was purified by affinity chromatography using a Ni-NTA column. The specific activity of the purified lysozyme was 34,000 U/mg using Micrococcus lysodeikticus as substrates. It exhibited antimicrobial activity toward M. lysodeikticus, as detected by growth inhibition on agar plate and turbidity assay, suggesting a potential application of A. japonicus lysozyme as an antimicrobial agent in A. japonicus aquaculture.

分类号： Q51

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