Molecular dynamic analysis of mutant Y195I α-cyclodextrin glycosyltransferase with switched product specificity from α-cyclodextrin to γ-cyclodextrin

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• 作者：Fangjin Chen ; Ting Xie ; Yang Yue ; Shijun Qian ; Yapeng Chao…
• 关键词：Cyclodextrin glucanotransferases ; Y195I α ; CGTase ; α ; CD ; γ ; CD ; Molecular dynamic simulation
• 刊名：Journal of Molecular Modeling
• 出版年：2015
• 出版时间：August 2015
• 年：2015
• 卷：21
• 期：8
• 全文大小：1,647 KB
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• 作者单位：Fangjin Chen (1)
  Ting Xie (2) (3)
  Yang Yue (2)
  Shijun Qian (2)
  Yapeng Chao (2)
  Jianfeng Pei (1) (4)
  
  1. Center for Quantitative Biology, AAIS, Peking University, Beijing, China, 100871
  2. State Key Laboratories of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China, 100101
  3. University of Chinese Academy of Sciences, Beijing, China, 100101
  4. State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, People’s Republic of China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Computer Applications in Chemistry
  Biomedicine
  Molecular Medicine
  Health Informatics and Administration
  Life Sciences
  Computer Application in Life Sciences
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：0948-5023

文摘

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.