title: Extracellular production and characterization of Streptomyces X-prolyl dipeptidyl aminopeptidase.
abstract: X-prolyl dipeptidyl aminopeptidases (X-PDAPs) are useful in various food industries. In this study, we performed sequence-based screening to obtain a stable X-PDAP enzyme from thermophilic Streptomyces strains. We found three genes that encoded X-PDAP from Streptomyces thermoluteus subsp. fuscus NBRC 14270 (14270 X-PDAP), Streptomyces thermocyaneoviolaceus NBRC 14271 (14271 X-PDAP), and Streptomyces thermocoerulescens NBRC 14273, which were subsequently cloned and sequenced. The deduced amino acid sequences of these genes showed high similarity, with ~80% identity with each other. The isolated X-PDAPs and an X-PDAP from Streptomyces coelicolor were expressed in Streptomyces lividans using a hyperexpression vector: pTONA5a. Among these genes, only 14270 and 14271 X-PDAPs caused overexpression and extracellular production without artificial signal peptides. We also characterized the biochemical properties of purified 14271 X-PDAP. In addition, we found that, in peptide synthesis via an aminolysis reaction, this enzyme recognized D-amino acid derivatives as acyl acceptors, similar to L-amino acid derivatives.
authors.affiliation: Okayama Prefectural Technology Center for Agriculture, Forestry, and Fisheries, Research Institute for Biological Sciences (RIBS), Okayama, Kibichuo-cho, Kaga-gun, Japan. hatanaka@bio-ribs.com
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hatanaka
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yamasato
authors.forename: Jiro
authors.initials: J
authors.lastname: Arima
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Usuki
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Yuya
authors.initials: Y
authors.lastname: Kumagai
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D004269:DNA, Bacterial / Q000378:metabolism; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000096:biosynthesis* / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D013302:Streptomyces / Q000201:enzymology* / Q000378:metabolism; D013696:Temperature
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C098392:PepX dipeptidyl aminopeptidase