Production, purification, and properties of an endoglucanase produced by the hyphomycete Chalara (Syn. Thielaviopsis) paradoxa CH32.
The hyphomycete Chalara (syn. Thielaviopsis) paradoxa produces endoglucanase activity during the late trophophase. The low molecular mass (35 kDa) endoglucanase purified from cultured broths works optimally at 37 degrees C and pH 5.0. The enzyme inactivates at pH below 3.0 and also at temperatures of 50 degrees C or higher, but it is stable at lower temperatures, including refrigeration temperature and freezing. The enzyme is inhibited by detergents, by EDTA, and by the divalent cations Hg(2+) and Ag(2+). It is also inhibited to some extent by 10 mM Zn(2+), Fe(2+), and Mg(2+), but it is stimulated by Mn(2+). Enzyme activity is not affected by reducing agents. In the presence of low concentrations of water miscible organic solvents (20%) endoglucanase activity is inhibited by 7% (for methanol) to 50% (for acetonitrile), and it is totally inhibited at higher solvent concentrations (50%). Enzyme activity is not affected by the water immiscible solvent ethyl acetate. Carboxymethylcellulose is the preferred substrate (K(m(app)) = 8.3 g/L; V(max(app)) = 1.1 microM/min). Hydrolysis of crystalline cellulosic substrates is very limited, but it is greatly enhanced by phosphoric acid swelling. The purified enzyme shows no activity toward disaccharides or aryl-glucosides. Its activity is inhibited by cellobiose.
Lucas
R
R
Microbiology Division, Faculty of Experimental Sciences, University of Jaén, 23071 Jaen, Spain.
Robles
A
A
García
M T
MT
Alvarez De Cienfuegos
G
G
Gálvez
A
A
D000097:Acetonitriles / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D002480:Cellulase / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005296:Ferrous Compounds / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008628:Mercury / Q000494:pharmacology; D000432:Methanol / Q000494:pharmacology; D003904:Mitosporic Fungi / Q000201:enzymology*; D008970:Molecular Weight; D012834:Silver / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature; D015032:Zinc / Q000494:pharmacology
D000097:Acetonitriles; D002413:Cations, Divalent; D003902:Detergents; D004791:Enzyme Inhibitors; D005296:Ferrous Compounds; D012834:Silver; D004492:Edetic Acid; D002480:Cellulase; D008628:Mercury; D008274:Magnesium; D015032:Zinc; D000432:Methanol; C032159:acetonitrile