Cloning of the Penicillium canescens endo-1,4-beta-glucanase gene egl3 and the characterization of the recombinant enzyme

• Published in: Prikladnaja biohimija i mikrobiologija Vol. 45; no. 2; p. 163
• Main Authors: Chulkin, A M, Loginov, D S, Vavilova, E A, Abaianova, A R, Zorov, I N, Kurzeev, S A, Koroleva, O V, Benevolenskiĭ, S V
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01.03.2009
• Subjects: Cellulase - biosynthesis; Cellulase - chemistry; Cellulase - genetics; Cellulase - isolation & purification; Cloning, Molecular - methods; Fungal Proteins - biosynthesis; Fungal Proteins - chemistry; Fungal Proteins - isolation & purification; Gene Expression; Hot Temperature; Hydrogen-Ion Concentration; Kinetics; Penicillium - enzymology; Penicillium - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification
• ISSN: 0555-1099

The gene egl3 of the filamentous fungus Penicillium canescens endo-1,4-beta-glucanase, belonging to family 12 glycosyl hydrolases, was cloned and sequenced. The gene was expressed in P. canescens under the control of the strong promoter of gene bgaS, coding for beta-galactosidase of this fungus, and efficient endoglucanase producer strains were obtained. The recombinant protein was isolated from the culture liquid of the producer strain EGL3-13 and purified to homogeneity; its specific activity was 31.7 IU; molecular weight, 26 kDa; and pH and temperature optimums, 3.2 and 54 degrees C, respectively. The Km and Vm values for CMC hydrolysis were determined; they amounted to 17.1 g/1 and 0.31 microM/(mg s), respectively.