Open AccessPurification, identification and molecular cloning of glycoside hydrolase family 15 glucoamylase from the brown‐rot basidiomycete Fomitopsis palustris
Author(s) -
Yoon JeongJun,
Igarashi Kiyohiko,
Kajisa Taira,
Samejima Masahiro
Publication year - 2006
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.1574-6968.2006.00279.x
Subject(s) - cellobiose , glycoside hydrolase , biochemistry , complementary dna , biology , hydrolase , peptide sequence , amino acid , enzyme , molecular cloning , cloning (programming) , cellulase , gene , computer science , programming language
The brown‐rot basidiomycete Fomitopsis palustris produces a major extracellular enzyme of 72 kDa when the fungus is incubated in cellulose culture with 0.2% cellobiose. This protein was purified by column chromatography, and the amino acid sequences of its proteolytic fragments were analyzed. The N‐terminal amino acid sequence of one of the fragments showed high identity with fungal glycoside hydrolase family 15 glucoamylases. As its kinetic efficiency increased in proportion to the degree of polymerization of the substrate, the protein was identified as a glucoamylase. A cDNA encoding the glucoamylase ( gla ) was cloned by reverse transcriptase PCR.