Open AccessProperties of yeast expressed Aspergillus nidulans chitin synthase B which is essential for hyphal growth 1
Author(s) -
Tatsuno Kenji,
YamadaOkabe Hisafumi,
Takagi Masamichi,
Arisawa Mikio,
Sudoh Masayuki
Publication year - 1997
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.1574-6968.1997.tb10341.x
Subject(s) - chitin synthase , aspergillus nidulans , chitin , saccharomyces cerevisiae , biochemistry , hypha , yeast , biology , enzyme , atp synthase , chemistry , microbiology and biotechnology , gene , mutant , chitosan
A complementary DNA of the Aspergillus nidulans chsB gene encoding chitin synthase, an essential gene for hyphal growth, was obtained by RT‐PCR and expressed in Saccharomyces cerevisiae by using the GAL1 promoter in a multicopy plasmid. The biochemical characteristics of chitin synthase B (ChsB) expressed in S. cerevisiae were examined. The chitin synthase B produced in galactose medium showed zymogenicity due to activation by trypsin treatment and required Mg 2+ ion to exert maximal activity. It was competitively inhibited by polyoxin D. The K i value of the inhibitor was 10 μM, and the K m for the substrate was 1.6 mM. The activity was enhanced by the addition of N ‐acetylglucosamine. The optimal pH is 7.5 when Mg 2+ is used. These characteristics are the same as those of other chitin synthases.