Enhancement of alpha-cyclodextrin product specificity by enriching histidines of alpha-cyclodextrin glucanotransferase at remote subsite-6

	Enhancement of alpha-cyclodextrin product specificity by enriching histidines of alpha-cyclodextrin glucanotransferase at remote subsite-6
	Yue, Yang 1,2; Song, Binghong 2; Xie, Ting 2; Sun, Yan 1; Chao, Yapeng 2; Qian, Shijun 2
刊名	PROCESS BIOCHEMISTRY
	2014-02-01
卷号	49 期号:2 页码:230-236
关键词	alpha-Cyclodextrin glucanotransferases alpha-Cyclodextrin Product specificity Site-directed mutagenesis Enzymatic properties
英文摘要	The industrial use of a-cyclodextrins (alpha-CDs) has increased because their solubility is higher than those of beta-CDs. However, improving the product specificity of alpha-cyclodextrin glucanotransferases (CGTases) remains unresolved. In this study, three mutants (Y167-deletion, Y167HH, and Y167HHH) were constructed at subsite 6 of alpha-CGTase to investigate the contribution of amino acid residue 167 to the cyclization ability of alpha-CD by comparing it with Tyr167His mutant a-CGTase (previously constructed based on the wild-type gene of Bacillus sp. 602-1). As expected, the alpha:beta ratio improved with increasing number of histidine along with residue 167. The Y167HHH mutant had the highest alpha:beta ratio of 13.2 and almost produced single type a-CDs. The Y167HHH mutant enzyme was subsequently purified to homogeneity. The enzymatic properties and the optimal condition of Y167HHH mutant in converting raw starch were also investigated. This study discusses product specificity improvement by inserting specific amino acid residues in the active groove. The results indicate that the histidine-rich mutant alpha-CGTase possessed better potential in producing alpha-CDs in an industrial scale. (c) 2013 Published by Elsevier Ltd.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Technology
类目[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
研究领域[WOS]	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
关键词[WOS]	CIRCULANS STRAIN 251 ; SWISS-MODEL ; PROTEIN-STRUCTURE ; GLYCOSYLTRANSFERASE ; ENVIRONMENT ; INHIBITION ; MECHANISM ; BINDING ; ENZYME
收录类别	SCI
语种	英语
WOS记录号	WOS:000332498200008
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1408]
专题	天津工业生物技术研究所_酶工程实验室马延和 _期刊论文
作者单位	1.Beihang Univ BUAA, Key Lab Biomech & Mechanobiol, Minist Educ, Sch Biol Sci & Med Engn, Beijing 100191, Peoples R China 2.Chinese Acad Sci, Inst Microbiol, Natl Engn Lab Ind Enzymes, State Key Labs Transducer Technol, Beijing 100101, Peoples R China
推荐引用方式 GB/T 7714	Yue, Yang,Song, Binghong,Xie, Ting,et al. Enhancement of alpha-cyclodextrin product specificity by enriching histidines of alpha-cyclodextrin glucanotransferase at remote subsite-6[J]. PROCESS BIOCHEMISTRY,2014,49(2):230-236.
APA	Yue, Yang,Song, Binghong,Xie, Ting,Sun, Yan,Chao, Yapeng,&Qian, Shijun.(2014).Enhancement of alpha-cyclodextrin product specificity by enriching histidines of alpha-cyclodextrin glucanotransferase at remote subsite-6.PROCESS BIOCHEMISTRY,49(2),230-236.
MLA	Yue, Yang,et al."Enhancement of alpha-cyclodextrin product specificity by enriching histidines of alpha-cyclodextrin glucanotransferase at remote subsite-6".PROCESS BIOCHEMISTRY 49.2(2014):230-236.