Optimized production of scygonadin in Pichia pastoris and analysis of its antimicrobial and antiviral activities

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	Optimized production of scygonadin in Pichia pastoris and analysis of its antimicrobial and antiviral activities
	Peng, Hui ; Liu, Hai-Peng ; Chen, Bei ; Hao, Hua ; Wang, Ke-Jian ; Liu HP(刘海鹏)
刊名	http://dx.doi.org/10.1016/j.pep.2011.11.008
	2012-03
关键词	HIGH-LEVEL EXPRESSION SPOT SYNDROME VIRUS ANTILIPOPOLYSACCHARIDE FACTOR ALF CRAB SCYLLA-PARAMAMOSAIN MUD CRAB MOLECULAR-CLONING GENE-EXPRESSION PACIFASTACUS-LENIUSCULUS ANTIBACTERIAL PROTEIN REPRODUCTIVE IMMUNITY
英文摘要	National Natural Science Foundation of China (NSFC) [40676083]; Public Science and Technology Research Funds Projects of Ocean, State Oceanic Administration of the People's Republic of China [201105027]; The crab antimicrobial peptide scygonadin is confirmed to have antimicrobial activity against bacteria and it is probably associated with the reproductive immunity in Scylla paramamosain. To obtain large quantity of scygonadin for further biological assays, a 306 bp cDNA sequence encoding the mature peptide of scygonadin was cloned into a secretion vector of pPIC9K, and a high-level of the recombinant scygonadin was achieved in Pichia pastoris. The optimal expression condition was determined as incubation with 0.5% methanol for 48 h at 28 degrees C under pH 6.0, and a total of 70 mg scygonadin was expressed in 1 L culture medium. The recombinant product was purified and 97% pure scygonadin was obtained using immobilized metal affinity chromatography with a yield of 46 mg/L. The recombinant scygonadin was confirmed using SDS-PAGE analysis and MS-fingerprinting. P. pastoris-derived scygonadin exhibited relatively higher antimicrobial activities against bacteria than Escherichia coli-derived scygonadin. The antimicrobial activity of the recombinant scygonadin against pathogenic Aeromonas hydrophila showed salt resistant and the killing kinetics of A. hydrophila was time dependent. Besides, the antiviral assay demonstrated that scygonadin could interfere with white spot syndrome virus (WSSV) replication in vitro-cultured crayfish haematopoietic (Hpt) cells. Taken together, this is the first report on the heterologous expression of scygonadin in P. pastoris, and P. pastoris is an effective expression system for producing large quantities of biological active scygonadin for both research and agricultural application. (C) 2011 Elsevier Inc. All rights reserved.
语种	英语
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/60112]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	Peng, Hui,Liu, Hai-Peng,Chen, Bei,et al. Optimized production of scygonadin in Pichia pastoris and analysis of its antimicrobial and antiviral activities[J]. http://dx.doi.org/10.1016/j.pep.2011.11.008,2012.
APA	Peng, Hui,Liu, Hai-Peng,Chen, Bei,Hao, Hua,Wang, Ke-Jian,&刘海鹏.(2012).Optimized production of scygonadin in Pichia pastoris and analysis of its antimicrobial and antiviral activities.http://dx.doi.org/10.1016/j.pep.2011.11.008.
MLA	Peng, Hui,et al."Optimized production of scygonadin in Pichia pastoris and analysis of its antimicrobial and antiviral activities".http://dx.doi.org/10.1016/j.pep.2011.11.008 (2012).