Single step immobilization of CMCase within agarose gel matrix: Kinetics and thermodynamic studies

In the current study, CMCase from Bacillus licheniformis KIBGE-IB2 was immobilized within the matrix of agarose gel through entrapment technique. Maximum immobilization yield (%) of the enzyme was obtained when 2.0 % agarose was used. The activation energy (E-a) of the enzyme increased from 16.38 to 44.08 kJ mol 1 after immobilization. Thermodynamic parameters such as activation energy of deactivation (Delta G(d)), enthalpy (Delta H-d) and entropy (Delta S-d) of deactivation, deactivation rate constant (K-d), half-life (t(1/2)), D-value and z-value were calculated for native/free and immobilized CMCase. The maximum reaction rate (V-max) of the native enzyme was found to be 8319.47 U ml(-1) min(-1), which reduced to 7218.1 U ml(-1) min(-1) after immobilization process. However, the Michaelis-Menten constant (K-m) value of the enzyme increased from 1.236 to 2.769 mg ml(-1) min(-1) after immobilization. Immobilized enzyme within agarose gel matrix support can be reuse up to eight reaction cycles. Broad stability profile and improved catalytic properties of the immobilized CMCase indicated that this enzyme can be a plausible candidate to be used in various industrial processes.

Automated summary

In this study, CMCase from Bacillus licheniformis KIBGE-IB2 was immobilized within agarose gel matrix, leading to improved catalytic properties with reduced maximum reaction rate and increased Michaelis-Menten constant. The immobilized enzyme showed enhanced stability and could be reused up to eight reaction cycles, indicating its potential application in various industrial processes.