Improved extracellular expression and high-cell-density fed-batch fermentation of chitosanase from Aspergillus Fumigatus in Escherichia coli

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	Improved extracellular expression and high-cell-density fed-batch fermentation of chitosanase from Aspergillus Fumigatus in Escherichia coli
	Huang, Liang 1; Wang, Qinhong 2; Jiang, Sijing 1; Zhou, Yuling 1; Zhang, Guimin 1; Ma, Yanhe 2
刊名	BIOPROCESS AND BIOSYSTEMS ENGINEERING
	2016-11-01
卷号	39 期号:11 页码:1679-1687
关键词	Chitosanase Secretory expression Fed-batch fermentation
英文摘要	Chitosanase (CSN) from Aspergillus fumigatus has good thermal stability, wide pH range duration, and effective hydrolysis for chitosan. Inhere, CSN was successfully expressed in Escherichia coli followed by extracellular secretion under the guidance of an N-terminal signal peptide PelB, which effectively prompted its secretion out of E. coli cells. To facilitate its later purification, N-terminal or C-terminal 6xHis epitope tag was added to the PelB-CSN protein complex. Our results indicated that PelB-CSN without 6xHis-tag (PelB-CSN) or with N-terminal 6xHis-tag (PelB-CSN-N) can both be effectively secreted into the medium, while CSN with 6xHis-tag anchored at C-terminus was expressed as inclusion bodies. Process optimization strategies were further developed to improve the secretion efficiency of recombinant PelB-CSN-N in E. coli. Under the induction of 10 g/L lactose in shake-flask culture, the extracellular activity of CSN reached 6015 U/mL at 25 A degrees C in TB medium containing 1 % glycine. Moreover, a fed-batch fermentation strategy for high-cell-density cultivation was applied in a 5-L fermenter, increasing the extracellular CSN activity to 14,000 U/mL in 2-day fermentation with the optimal addition of lactose and glycine.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Technology
类目[WOS]	Biotechnology & Applied Microbiology ; Engineering, Chemical
研究领域[WOS]	Biotechnology & Applied Microbiology ; Engineering
关键词[WOS]	FAMILY 75 CHITOSANASE ; ENHANCED SECRETION ; SIGNAL PEPTIDE ; BL21 DE3 ; OLIGOSACCHARIDES ; OPTIMIZATION ; GLYCINE ; LYASE ; MODE
收录类别	SCI
语种	英语
WOS记录号	WOS:000384354100004
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2935]
专题	天津工业生物技术研究所_进化代谢工程和分子进化研究组王钦宏_期刊论文
作者单位	1.Hubei Univ, Hubei Collaborat Innovat Ctr Green Transformat Bi, Coll Life Sci, Wuhan 430062, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China
推荐引用方式 GB/T 7714	Huang, Liang,Wang, Qinhong,Jiang, Sijing,et al. Improved extracellular expression and high-cell-density fed-batch fermentation of chitosanase from Aspergillus Fumigatus in Escherichia coli[J]. BIOPROCESS AND BIOSYSTEMS ENGINEERING,2016,39(11):1679-1687.
APA	Huang, Liang,Wang, Qinhong,Jiang, Sijing,Zhou, Yuling,Zhang, Guimin,&Ma, Yanhe.(2016).Improved extracellular expression and high-cell-density fed-batch fermentation of chitosanase from Aspergillus Fumigatus in Escherichia coli.BIOPROCESS AND BIOSYSTEMS ENGINEERING,39(11),1679-1687.
MLA	Huang, Liang,et al."Improved extracellular expression and high-cell-density fed-batch fermentation of chitosanase from Aspergillus Fumigatus in Escherichia coli".BIOPROCESS AND BIOSYSTEMS ENGINEERING 39.11(2016):1679-1687.