Open AccessCloning and expression of a β‐1,4‐endoglucanase gene from Cellulomonas sp. CelB7 in Escherichia coli ; purification and characterization of the recombinant enzyme
Author(s) -
Fülöp László,
Trân Son Lam Phan,
Prágai Zoltán,
Felföldi Ferenc,
Ponyi Tamás
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.tb08600.x
Subject(s) - cellulase , escherichia coli , recombinant dna , molecular cloning , gene , biochemistry , enzyme , biology , microbiology and biotechnology , cloning (programming) , fast protein liquid chromatography , chemistry , gene expression , computer science , programming language
A gene library of a newly isolated Cellulomonas sp. strain was constructed in Escherichia coli and clones were screened for endoglucanase activity using dye‐labelled carboxymethylcellulose. Seventeen clones were isolated that carried DNA inserts coding for endoglucanase enzymes. Of the 17 clones, one carrying the gene cegA , was further characterized. The recombinant endoglucanase was purified by FPLC. The endoglucanase was active against carboxymethylcellulose, lichenin and also degraded crystalline cellulose and birchwood xylan. The molecular mass of the enzyme (36 kDa), and its pH (7.4) and temperature (35 °C) optima were determined.