caa a22 4500 605512957 CHVBK 20210128100658.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00894-015-2734-x doi (NATIONALLICENCE)springer-10.1007/s00894-015-2734-x Molecular dynamic analysis of mutant Y195I α-cyclodextrin glycosyltransferase with switched product specificity from α-cyclodextrin to γ-cyclodextrin [Elektronische Daten] [Fangjin Chen, Ting Xie, Yang Yue, Shijun Qian, Yapeng Chao, Jianfeng Pei] Alpha-cyclodextrin (α-CD) glycosyltransferase (α-CGTase) can convert starch into α-CD blended with various proportions of β-cyclodextrin (β-CD) and/or γ-cyclodextrin (γ-CD). In this study, we verified the catalytic characteristics of purified Y195I α-CGTase and elucidated the mechanism of action with molecular dynamic (MD) simulations. We found that purified Y195I α-CGTase produced less α-CD, slightly more β-CD, and significantly more γ-CD than wild-type α-CGTase. Correspondingly, α-CD-based K m values increased, and β-CD- and γ-CD-based K m values decreased. MD simulation studies revealed that the dynamic trajectories of the substrate oligosaccharide chain in the mutant CGTase binding site were significantly different from those in the wild-type enzyme, with reduced hydrophobic interaction, finally resulting in different product specificity and more γ-CD formation. Springer-Verlag Berlin Heidelberg, 2015 Cyclodextrin glucanotransferases nationallicence Y195I α-CGTase nationallicence α-CD nationallicence γ-CD nationallicence Molecular dynamic simulation nationallicence Chen Fangjin Center for Quantitative Biology, AAIS, Peking University, 100871, Beijing, China aut Xie Ting State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Yue Yang State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Qian Shijun State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Chao Yapeng State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Pei Jianfeng Center for Quantitative Biology, AAIS, Peking University, 100871, Beijing, China aut Journal of Molecular Modeling Springer Berlin Heidelberg 21/8(2015-08-01), 1-9 1610-2940 21:8<1 2015 21 894 https://doi.org/10.1007/s00894-015-2734-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00894-015-2734-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen Fangjin Center for Quantitative Biology, AAIS, Peking University, 100871, Beijing, China aut NATIONALLICENCE 700 1- Xie Ting State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Yue Yang State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Qian Shijun State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Chao Yapeng State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Pei Jianfeng Center for Quantitative Biology, AAIS, Peking University, 100871, Beijing, China aut NATIONALLICENCE 773 0- Journal of Molecular Modeling Springer Berlin Heidelberg 21/8(2015-08-01), 1-9 1610-2940 21:8<1 2015 21 894