title: Catalytic and thermodynamic characterization of endoglucanase (CMCase) from Aspergillus oryzae cmc-1.
abstract: Monomeric extracellular endoglucanase (25 kDa) of transgenic koji (Aspergillus oryzae cmc-1) produced under submerged growth condition (7.5 U mg(-1) protein) was purified to homogeneity level by ammonium sulfate precipitation and various column chromatography on fast protein liquid chromatography system. Activation energy for carboxymethylcellulose (CMC) hydrolysis was 3.32 kJ mol(-1) at optimum temperature (55 degrees C), and its temperature quotient (Q (10)) was 1.0. The enzyme was stable over a pH range of 4.1-5.3 and gave maximum activity at pH 4.4. V (max) for CMC hydrolysis was 854 U mg(-1) protein and K (m) was 20 mg CMC ml(-1). The turnover (k (cat)) was 356 s(-1). The pK (a1) and pK (a2) of ionisable groups of active site controlling V (max) were 3.9 and 6.25, respectively. Thermodynamic parameters for CMC hydrolysis were as follows: DeltaH* = 0.59 kJ mol(-1), DeltaG* = 64.57 kJ mol(-1) and DeltaS* = -195.05 J mol(-1) K(-1), respectively. Activation energy for irreversible inactivation 'E (a(d))' of the endoglucanase was 378 kJ mol(-1), whereas enthalpy (DeltaH*), Gibbs free energy (DeltaG*) and entropy (DeltaS*) of activation at 44 degrees C were 375.36 kJ mol(-1), 111.36 kJ mol(-1) and 833.06 J mol(-1) K(-1), respectively.
authors.affiliation: Enzyme Engineering Group, Industrial Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, P.O. Box 577, Jhang Road, Faisalabad, Pakistan.
authors.forename: Muhammad Rizwan
authors.initials: MR
authors.lastname: Javed
authors.forename: Muhammad Hamid
authors.initials: MH
authors.lastname: Rashid
authors.forename: Habibullah
authors.initials: H
authors.lastname: Nadeem
authors.forename: Muhammad
authors.initials: M
authors.lastname: Riaz
authors.forename: Raheela
authors.initials: R
authors.lastname: Perveen
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology*; D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002384:Catalysis; D002480:Cellulase / Q000302:isolation & purification / Q000378:metabolism*; D019277:Entropy; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D013816:Thermodynamics
chemical_list: D002480:Cellulase; C019826:carboxymethylcellulase; D002266:Carboxymethylcellulose Sodium