Truncation of the unique N-terminal domain improved the thermos-stability and specific activity of alkaline alpha-amylase Amy703

CORC > 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

	Truncation of the unique N-terminal domain improved the thermos-stability and specific activity of alkaline alpha-amylase Amy703
	Lu, Zhenghui 1; Wang, Qinhong 2; Jiang, Sijing 1; Zhang, Guimin 1; Ma, Yanhe 2
刊名	SCIENTIFIC REPORTS
	2016-03-01
卷号	6
英文摘要	High pH condition is of special interest for the potential applications of alkaline alpha-amylase in textile and detergent industries. Thus, there is a continuous demand to improve the amylase's properties to meet the requirements set by specific applications. Here we reported the systematic study of modular domain engineering to improve the specific activity and stability of the alkaline alpha-amylase from Bacillus pseudofirmus 703. The specific activity of the N-terminal domain truncated mutant (N-Amy) increased by similar to 35-fold with a significantly improved thermo-stability. Kinetic analysis demonstrated that the K-cat and K-cat/K-m of N-Amy were enhanced by 1300-fold and 425.7-fold, respectively, representing the largest catalytic activity improvement of the engineered alpha-amylases through the methods of domain deletion, fusion or swapping. In addition, different from the wild-type Amy703, no exogenous Ca2+ were required for N-Amy to maintain its full catalytic activity, implying its superior potential for many industrial processes. Circular dichroism analysis and structure modeling revealed that the increased compactness and a-helical content were the main contributors for the improved thermo-stability of N-Amy, while the improved catalytic efficiency was mainly attributed by the increased conformational flexibility around the active center.
WOS标题词	Science & Technology
类目[WOS]	Multidisciplinary Sciences
研究领域[WOS]	Science & Technology - Other Topics
关键词[WOS]	DIRECTED EVOLUTION ; RATIONAL DESIGN ; FAMILY ; BIOCATALYSTS ; ENZYMES ; PULLULANASE ; PROTEINS ; SEQUENCE ; BINDING
收录类别	SCI
语种	英语
WOS记录号	WOS:000371037400001
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1540]
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	Lu, Zhenghui,Wang, Qinhong,Jiang, Sijing,et al. Truncation of the unique N-terminal domain improved the thermos-stability and specific activity of alkaline alpha-amylase Amy703[J]. SCIENTIFIC REPORTS,2016,6.
APA	Lu, Zhenghui,Wang, Qinhong,Jiang, Sijing,Zhang, Guimin,&Ma, Yanhe.(2016).Truncation of the unique N-terminal domain improved the thermos-stability and specific activity of alkaline alpha-amylase Amy703.SCIENTIFIC REPORTS,6.
MLA	Lu, Zhenghui,et al."Truncation of the unique N-terminal domain improved the thermos-stability and specific activity of alkaline alpha-amylase Amy703".SCIENTIFIC REPORTS 6(2016).