caa a22 4500 605506329 CHVBK 20210128100626.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-014-6304-4 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6304-4 Design of N -acyl homoserine lactonase with high substrate specificity by a rational approach [Elektronische Daten] [Hyun-Ho Kyeong, Jin-Hyun Kim, Hak-Sung Kim] 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. Springer-Verlag Berlin Heidelberg, 2014 Quorum quenching nationallicence Acyl homoserine lactonase nationallicence Substrate preference nationallicence Rational design nationallicence Kyeong Hyun-Ho Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 305-701, Daejeon, Korea aut Kim Jin-Hyun LG Household & Health Care Ltd., R&D Institute, 305-343, Daejeon, Korea aut Kim Hak-Sung Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 305-701, Daejeon, Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4735-4742 0175-7598 99:11<4735 2015 99 253 https://doi.org/10.1007/s00253-014-6304-4 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/s00253-014-6304-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kyeong Hyun-Ho Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 305-701, Daejeon, Korea aut NATIONALLICENCE 700 1- Kim Jin-Hyun LG Household & Health Care Ltd., R&D Institute, 305-343, Daejeon, Korea aut NATIONALLICENCE 700 1- Kim Hak-Sung Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 305-701, Daejeon, Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4735-4742 0175-7598 99:11<4735 2015 99 253