Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp in Escherichia coli

doi:10.1007/s12088-014-0505-5

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	Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp in Escherichia coli
	Sun, Yuying 1,2,3; Zhang, Jiquan 2; Wang, Shujun 1,3
	2015-06-01
发表期刊	INDIAN JOURNAL OF MICROBIOLOGY
卷号	55 期号:2 页码:194-199
文章类型	Article
摘要	A recombinant expression vector, pCT7-CHISP6H, was constructed for the secretory expression of mature peptide of chitosanase (mMschito) from Microbacterium sp. OU01. The vector contains several elements, including T7 promoter, signal peptide sequence of mschito, 6 x His-tag sequence and PmaCI restriction enzyme cloning site. In pCT7-CHISP6H, mMschito was fused into signal peptide sequence of mschito gene to construct recombinant plasmid pCT7-CHISP6H-mMschito. The recombinant plasmid was transformed into Escherichia coli BL21(DE3) and then expressed. The recombinant protein was secreted into the Luria-Bertani broth and the chitosanase activity in supernatant of the culture could reach up to 67.56 U/mL. The rmMschito in the broth supernatant was purified using HisTrap (TM) FF Crude column and the purified rmMschito was shown to be apparent homogeneity by 12 % SDS-PAGE analysis. Detected by 4700 MALDI-TOF-TOF-MS, the molecular weight of the purified rmMschito was 26,758.1875 and it was consistent with the predicted molecular weight. Chitosan (degree of deacetylation of 99 %) was mostly hydrolyzed into chitopentaose, chitotriose, and chitobiose by the purified rmMschito.
关键词	Heterologous Secretion Microbacterium Sp Ou01 Recombinant Expression Vector Chitosanase
DOI	10.1007/s12088-014-0505-5
收录类别	SCI
语种	英语
WOS记录号	WOS:000351468900009
引用统计	被引频次：18[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/25443
专题	实验海洋生物学重点实验室
作者单位	1.Huaihai Inst Technol, Jiangsu Key Lab Marine Pharmaceut Compound Screen, Lianyungang 222005, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 3.Huaihai Inst Technol, Coinnovat Ctr Jiangsu Marine Bioind Technol, Lianyungang 222005, Peoples R China
第一作者单位	实验海洋生物学重点实验室
推荐引用方式 GB/T 7714	Sun, Yuying,Zhang, Jiquan,Wang, Shujun. Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp in Escherichia coli[J]. INDIAN JOURNAL OF MICROBIOLOGY,2015,55(2):194-199.
APA	Sun, Yuying,Zhang, Jiquan,&Wang, Shujun.(2015).Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp in Escherichia coli.INDIAN JOURNAL OF MICROBIOLOGY,55(2),194-199.
MLA	Sun, Yuying,et al."Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp in Escherichia coli".INDIAN JOURNAL OF MICROBIOLOGY 55.2(2015):194-199.

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