title: Purification and characterization of aqualysin I (a thermophilic alkaline serine protease) produced by Thermus aquaticus YT-1.
abstract: Aqualysin I is an alkaline serine protease which is secreted into the culture medium by Thermus aquaticus YT-1. Aqualysin I was purified, and its apparent relative molecular mass was determined to be 28 500. The enzyme contained four Cys residues (probably as two cystines), and its amino acids composition was similar to those of cysteine-containing serine proteases (proteinase K, etc.) as well as those of subtilisins. The NH2-terminal sequence of aqualysin I showed homology with those of the microbial serine proteases. The optimum pH for the proteolytic activity of aqualysin I was around 10.0. Ca2+ stabilized the enzyme to heat treatment, and the maximum proteolytic activity was observed at 80 degrees C. Aqualysin I was stable to denaturing reagents (7 M urea, 6 M guanidine.HCl and 1% SDS) at 23 degrees C for 24 h. The enzyme hydrolyzed the ester bond of an alanine ester and succinyl-Ala-Ala-Ala p-nitroanilide, a synthetic substrate for mammalian elastase. The cleavage sites for aqualysin I in oxidized insulin B chain were not specific when it was digested completely.
authors.affiliation: Department of Agricultural Chemistry, University of Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Matsuzawa
authors.forename: K
authors.initials: K
authors.lastname: Tokugawa
authors.forename: M
authors.initials: M
authors.lastname: Hamaoki
authors.forename: M
authors.initials: M
authors.lastname: Mizoguchi
authors.forename: H
authors.initials: H
authors.lastname: Taguchi
authors.forename: I
authors.initials: I
authors.lastname: Terada
authors.forename: S T
authors.initials: ST
authors.lastname: Kwon
authors.forename: T
authors.initials: T
authors.lastname: Ohta
mesh_terms: D000468:Alkalies; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002384:Catalysis; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000302:isolation & purification*; D013696:Temperature; D013824:Thermus / Q000201:enzymology*
chemical_list: D000468:Alkalies; D000596:Amino Acids; D012697:Serine Endopeptidases; C055102:aqualysin I; D010196:Pancreatic Elastase