Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948

doi:10.1016/j.pep.2011.01.009

	Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948
	Cui, Shuo-shuo ; Lin, Xue-zheng 1; Shen, Ji-hong
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2011-06
卷号	77 期号:2 页码:166-172
关键词	Cold-active lipase Molecular chaperone Co-expression Soluble expression
ISSN号	1046-5928
DOI	10.1016/j.pep.2011.01.009
英文摘要	The cold-active lipase gene Lip-948, cloned from Antarctic psychrotrophic bacterium Psychrobacter sp. G, was ligated into plasmid pColdI. The recombinant plasmid pColdI + Lip-948 was then transformed into Escherichia coli BL21. SDS-PAGE analysis showed that there was substantive expression of lipase LIP-948 in E. coli with a yield of about 39% of total protein, most of which was present in the inclusion body. The soluble protein LIP-948 only consisted of 1.7% of total LIP-948 with a specific activity of 66.51 U/mg. Co-expression of molecular chaperones with the pColdI + Lip-948 were also carried out. The results showed that co-expression of different chaperones led to an increase or decrease in the formation of soluble LIP-948 in varying degrees. Co-expression of pColdI + Lip-948 with chaperone pTf16 and pGro7 decreased the amount of soluble LIP-948, while the soluble expression was enhanced when pColdI + Lip-948 was co-expressed with "chaperone team" plasmids (pKJE7, pG-Tf2, pG-KJE8), respectively. LIP-948 was most efficiently expressed in soluble form when it was co-expressed with pG-KJE8, which was up to 19.8% of intracellular soluble proteins and with a specific activity of 108.77 U/mg. The soluble LIP-948 was purified with amylase affinity chromatography and its enzymatic characters were studied. The optimal temperature and pH of LIP-948 was 35 degrees C and 8, respectively. The activity of LIP-948 dropped dramatically after incubation at 50 degrees C for 15 min and was enhanced by Sr2+, Ca2+. It preferentially hydrolyzed 4-nitrophenyl esters with the shorter carbon chain. (c) 2011 Elsevier Inc. All rights reserved.
资助项目	Basic Scientific Research Foundation of FIO, SOA[2007T11]
WOS关键词	NEWLY SYNTHESIZED PROTEINS ; ESCHERICHIA-COLI ; TRIGGER FACTOR ; RECOMBINANT PROTEINS ; FUNCTIONAL EXPRESSION ; ADAPTED ENZYMES ; TEMPERATURE ; PSEUDOMONAS ; CLONING ; AGGREGATION
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000288591900006
内容类型	期刊论文
源URL	[http://ir.fio.com.cn:8080/handle/2SI8HI0U/27161]
专题	自然资源部第一海洋研究所
通讯作者	Lin, Xue-zheng
作者单位	1.SOA, Inst Oceanog 1, Qingdao 266061, Peoples R China 2.SOA, Key Lab Marine Bioact Substances, Qingdao 266061, Peoples R China
推荐引用方式 GB/T 7714	Cui, Shuo-shuo,Lin, Xue-zheng,Shen, Ji-hong. Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948[J]. PROTEIN EXPRESSION AND PURIFICATION,2011,77(2):166-172.
APA	Cui, Shuo-shuo,Lin, Xue-zheng,&Shen, Ji-hong.(2011).Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948.PROTEIN EXPRESSION AND PURIFICATION,77(2),166-172.
MLA	Cui, Shuo-shuo,et al."Effects of co-expression of molecular chaperones on heterologous soluble expression of the cold-active lipase Lip-948".PROTEIN EXPRESSION AND PURIFICATION 77.2(2011):166-172.