Isolation of a chymotrypsinogen B-like enzyme from the archaeon Natronomonas pharaonis and other halobacteria |
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Authors: | H Stan-Lotter Edith Doppler Marina Jarosch Christian Radax Claudia Gruber Ken-ichi Inatomi |
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Institution: | Institute of Genetics and General Biology, Hellbrunnerstra?e 34, A-5020 Salzburg, Austria Tel. +43-662-8044-5756; Fax +43-662-8044-144 e-mail: helga.stan-lotter@sbg.ac.at, AT Zentrum für Ultrastrukturforschung, Gregor-Mendel-Str. 33, A-1180 Vienna, Austria, AT Advanced Technical R&D Center, Mitsubishi Electrical Company, Hyogo, Japan, JP
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Abstract: | A protease of a molecular mass of approximately 30 kDa 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°C. The enzyme was nearly equally active over the range of salt concentration from 0.5 to 4 M (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.
Received: September 12, 1998 / Accepted: December 15, 1998 |
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Keywords: | Natronomonas pharaonis Natronobacteria Archaea Serine protease Chymotrypsinogen Evolution |
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