Design of N-acyl homoserine lactonase with high substrate specificity by a rational approach

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• 作者：Hyun-Ho Kyeong ; Jin-Hyun Kim ; Hak-Sung Kim
• 关键词：Quorum quenching ; Acyl homoserine lactonase ; Substrate preference ; Rational design
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：June 2015
• 年：2015
• 卷：99
• 期：11
• 页码：4735-4742
• 全文大小：2,108 KB
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• 作者单位：Hyun-Ho Kyeong (1)
  Jin-Hyun Kim (2)
  Hak-Sung Kim (1)
  
  1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea
  2. LG Household & Health Care Ltd., R&D Institute, Daejeon, 305-343, Korea
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

N-Acyl homoserine lactone (AHL) is a major quorum-sensing signaling molecule in many bacterial species. Quorum-quenching (QQ) enzymes, which degrade such signaling molecules, have attracted much attention as an approach to controlling and preventing bacterial virulence and pathogenesis. However, naturally occurring QQ enzymes show a broad substrate spectrum, raising the concern of unintentionally attenuating beneficial effects by symbiotic bacteria. Here we report the rational design of acyl homoserine lactonase with high substrate specificity. Through docking analysis, we identified three key residues which play a key role in the substrate preference of the enzyme. The key residues were changed in a way that increases hydrophobic contact with a substrate having a short acyl chain (C4-AHL) while generating steric clashes with that containing a long acyl chain (C12-AHL). The resulting mutants exhibited a significantly shifted preference toward a substrate with a short acyl chain. Molecular dynamics simulations suggested that the mutations affect the behavior of a flexible loop, allowing tighter binding of a substrate with a short acyl chain.