title: Mutant form C115H of Clostridium sporogenes methionine γ-lyase efficiently cleaves S-Alk(en)yl-l-cysteine sulfoxides to antibacterial thiosulfinates.
abstract: Pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) catalyzes the β-elimination reaction of S-alk(en)yl-l-cysteine sulfoxides to thiosulfinates, which possess antimicrobial activity. Partial inactivation of the enzyme in the course of the reaction occurs due to oxidation of active site cysteine 115 conserved in bacterial MGLs. In this work, the C115H mutant form of Clostridium sporogenes MGL was prepared and the steady-state kinetic parameters of the enzyme were determined. The substitution results in an increase in the catalytic efficiency of the mutant form towards S-substituted l-cysteine sulfoxides compared to the wild type enzyme. We used a sulfoxide/enzyme system to generate antibacterial activity in situ. Two-component systems composed of the mutant enzyme and three S-substituted l-cysteine sulfoxides were demonstrated to be effective against Gram-positive and Gram-negative bacteria and three clinical isolates from mice. © 2016 IUBMB Life, 68(10):830-835, 2016.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Vitalia V
authors.initials: VV
authors.lastname: Kulikova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Natalya V
authors.initials: NV
authors.lastname: Anufrieva
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Revtovich
authors.affiliation: Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow, Russia.
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Chernov
authors.affiliation: Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow, Russia.
authors.forename: Georgii B
authors.initials: GB
authors.lastname: Telegin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Elena A
authors.initials: EA
authors.lastname: Morozova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia. tvd@eimb.ru, tvdemidkina@yandex.ru.
authors.forename: Tatyana V
authors.initials: TV
authors.lastname: Demidkina
mesh_terms: D000900:Anti-Bacterial Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D055162:Biocatalysis; D013437:Carbon-Sulfur Lyases / Q000737:chemistry* / Q000235:genetics; D003013:Clostridium / Q000201:enzymology; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry*; D052978:Disk Diffusion Antimicrobial Tests; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D013454:Sulfoxides / Q000737:chemistry; D013886:Thiosulfonic Acids / Q000138:chemical synthesis* / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D013454:Sulfoxides; D013886:Thiosulfonic Acids; D013437:Carbon-Sulfur Lyases; C021558:L-methionine gamma-lyase; D003545:Cysteine
keywords: C115H mutant form; antibacterial activity; methionine γ-lyase; sulfoxides; thiosulfinates