title: Purification and characterization of alkaline serine protease from an alkalophilic Streptomyces sp.
abstract: SAP, an extracellular alkaline serine protease produced by Streptomyces sp. YSA-130, was purified to homogeneity by CM-Sephadex column chromatography and crystallization. The enzyme was a monomeric protein with a molecular weight of 19,000 as estimated by SDS-PAGE and gel filtration. The amino acid composition and amino-terminal sequence of SAP were similar to those of other bacterial serine proteases, i.e., Streptomyces griseus proteases A and B, Lysobacter enzymogenes alpha-lytic protease and Nocardiopsis dassonvillei subsp. prasina OPC-210 alkaline serine protease NDP-I. The optimum temperature and pH for the enzyme activity were 60 degrees C and 11.5. The enzyme was stable up 50 degrees C, and between pHs 4 and 12. The activity was inhibited by Ag+, Hg2+, Co2+, sodium dodecyl sulfate. N-bromosuccinimide, diisopropyl phosphorofluoridate (DFP), 2,3-butanedione, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), iodoacetate, N-ethylmaleimide (NEM), phenylmethanesulfonyl fluoride (PMSF), and phenylglyoxal.
authors.affiliation: Department of Food and Biotechnology, College of Engineering, Yonsei University, Seoul, Korea.
authors.forename: D Y
authors.initials: DY
authors.lastname: Yum
authors.forename: H C
authors.initials: HC
authors.lastname: Chung
authors.forename: D H
authors.initials: DH
authors.lastname: Bai
authors.forename: D H
authors.initials: DH
authors.lastname: Oh
authors.forename: J H
authors.initials: JH
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012697:Serine Endopeptidases / Q000032:analysis / Q000302:isolation & purification*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013302:Streptomyces / Q000201:enzymology*; D013696:Temperature
chemical_list: D000596:Amino Acids; D008670:Metals; D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases