Cloning and expression of a β-1,4-endoglucanase gene from Cellulomonas sp. CelB7 in Escherichia coli; purification and characterization of the recombinant enzyme |
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| Authors: | Lá szló Fü lö p,Son Lam Phan Trâ n,Zoltá n Prá gai,Ferenc Felfö ldi,tamá s Ponyi |
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| Affiliation: | Department of Chemistry and Biochemistry, University of Agricultural Sciences, GödöllöH-2103, Hungary; Department of Biotechnology and Molecular Genetics, University of Agricultural Sciences, GödöllöH-2103, Hungary; Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Budapest H-1518, Hungary |
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| Abstract: | Abstract 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. |
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| Keywords: | β-1,4-Endoglucanase Cellulomonas Cloning Enzyme assay Purification |
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