title: Isolation of a chymotrypsinogen B-like enzyme from the archaeon Natronomonas pharaonis and other halobacteria.
abstract: A protease of a molecular mass of approximately 30kDa was isolated and purified from the haloalkaliphilic archaeon Natronomonas (formerly Natronobacterium) pharaonis. The enzyme hydrolyzed synthetic peptides, preferentially at the carboxyl terminus of phenylalanine or leucine, as well as large proteins. Hydrolysis occurred over the range of pH from 6 to 12, with an optimum at pH 10. The temperature optimum was 61 degrees C. The enzyme was nearly equally active over the range of salt concentration from 0.5 to 4M (NaCl or KCl). A strong cross-reaction with a polyclonal antiserum against human chymotrypsin was observed. Enzymatic activity was inhibited by typical serine protease inhibitors. There was significant homology between N-terminal and internal sequences from autolytic fragments and the sequence of bovine chymotrypsinogen B; the overall amino acid composition was similar to that of vertebrate chymotrypsinogens. Evidence for a zymogen-like processing of the protease was obtained. Cell extracts from other halobacteria exhibited similar proteolytic activity and immunoreactivity. The data suggested a widespread distribution of a chymotrypsinogen B-like protease among halo- and haloalkaliphilic Archaea.
authors.affiliation: Institute of Genetics and General Biology, Salzburg, Austria. helga.stan-lotter@sbg.ac.at
authors.forename: H
authors.initials: H
authors.lastname: Stan-Lotter
authors.forename: E
authors.initials: E
authors.lastname: Doppler
authors.forename: M
authors.initials: M
authors.lastname: Jarosch
authors.forename: C
authors.initials: C
authors.lastname: Radax
authors.forename: C
authors.initials: C
authors.lastname: Gruber
authors.forename: K I
authors.initials: KI
authors.lastname: Inatomi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002919:Chymotrypsinogen / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010450:Endopeptidases / Q000302:isolation & purification; D004792:Enzyme Precursors; D006801:Humans; D008969:Molecular Sequence Data; D019633:Natronobacterium / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
chemical_list: D004792:Enzyme Precursors; D002919:Chymotrypsinogen; D010450:Endopeptidases