title: Peptide bond formation by aminolysin-A catalysis: a simple approach to enzymatic synthesis of diverse short oligopeptides and biologically active puromycins.
abstract: A new S9 family aminopeptidase derived from the actinobacterial thermophile Acidothermus cellulolyticus was cloned and engineered into a transaminopeptidase by site-directed mutagenesis of catalytic Ser(491) into Cys. The engineered biocatalyst, designated aminolysin-A, can catalyze the formation of peptide bonds to give linear homo-oligopeptides, hetero-dipeptides, and cyclic dipeptides using cost-effective substrates in a one-pot reaction. Aminolysin-A can recognize several C-terminal-modified amino acids, including the l- and d-forms, as acyl donors as well as free amines, including amino acids and puromycin aminonucleoside, as acyl acceptors. The absence of amino acid esters prevents the formation of peptides; therefore, the reaction mechanism involves aminolysis and not a reverse reaction of hydrolysis. The aminolysin system will be a beneficial tool for the preparation of structurally diverse peptide mimetics by a simple approach.
authors.affiliation: Okayama Prefectural Technology Center for Agriculture, Forestry and Fisheries, Research Institute for Biological Sciences (RIBS), 7549-1 Kibichuo-cho, Kaga-gun, Okayama 716-1241, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Usuki
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Jiro
authors.initials: J
authors.lastname: Arima
authors.forename: Masaki
authors.initials: M
authors.lastname: Iwabuchi
authors.forename: Shozo
authors.initials: S
authors.lastname: Miyoshi
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Nitoda
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hatanaka
mesh_terms: D000192:Actinomycetales / Q000201:enzymology*; D000626:Aminopeptidases / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D055162:Biocatalysis; D050296:Microbial Viability / Q000187:drug effects; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D010802:Phylogeny; D011691:Puromycin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D009842:Oligopeptides; D011691:Puromycin; D000626:Aminopeptidases