Open AccessCloning and sequencing of a dextranase‐encoding cDNA from Penicillium minioluteum
Author(s) -
Garcia Bianca,
Margolles Emilio,
Roca Hernan,
Mateu Dania,
Raices Manuel,
Gonzales Maria Elena,
Herrera Luis,
Delgado Julio
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.tb08477.x
Subject(s) - dextranase , complementary dna , microbiology and biotechnology , biology , cdna library , molecular cloning , escherichia coli , peptide sequence , biochemistry , gene , enzyme
Abstract A cDNA from Penicillium minioluteum HI‐4 encoding a dextranase (1,6‐α‐glucan hydrolase, EC 3.2.1.11) was isolated and characterized. cDNA clones corresponding to genes expressed in dextran‐induced cultures were identified by differential hybridization. Southern hybridization and restriction mapping analysis of selected clones revealed four different groups of cDNAs. The dextranase cDNA was identified after expressing a cDNA fragment from each of the isolated groups of cDNA clones in the Escherichia coli T7 system. The expression of a 2 kb cDNA fragment in E. coli led to the production of a 67 kDa protein which was recognized by an anti‐dextranase polyclonal antibody. The cDNA contains 2109 bp plus a poly(A) tail, coding for a protein of 608 amino acids, including 20 N‐terminal amino acid residues which might correspond to a signal peptide. There was 29% sequence identity between the P. minioluteum dextranase and the dextranase from Arthrobacter sp. CB‐8.