Open AccessPurification and characteristics of cytosolic chitinase from Piromyces communis OTS1
Author(s) -
Sakurada Masaru,
Morgavi Diego P.,
Komatani Kenji,
Tomita Yoshifumi,
Onodera Ryoji
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08085.x
Subject(s) - chromatofocusing , chitinase , biochemistry , chitin , molecular mass , biology , affinity chromatography , size exclusion chromatography , cytosol , enzyme , glucanase , chromatography , chemistry , chitosan
A chitinase was purified from the cytosolic fraction of the anaerobic rumen fungus Piromyces communis OTS1 by affinity chromatography using regenerated chitin, gel filtration and chromatofocusing. The chitinase was most active at pH 6.2 and at 60 °C in a 20‐min assay. The molecular mass of the purified protein was estimated by SDS‐PAGE to be 42 kDa and its pI was 4.9. The enzyme activity, which was of the ‘endo’ type, was inhibited by A + , Hg 2+ and allosamidin. N ‐Acetyl‐ β ‐glucosaminidase and ‘exo’ type chitinase activity were absent from the purified preparation.