Background: In this paper, cetyltrimethylammonium bromide (CTAB)-KOH modified biochar (CKMB) was used to immobilize laccase by adsorption method ([email protected]). Based on the results of the single-factor experiments, the optimal loading conditions of [email protected] were studied with the assistance of Design-Expert 12 and response surface methods.
Results: The predicted optimal experimental conditions were laccase dosage 1.78 mg/mL, pH 3.1 and 312 K. Under these conditions, the activity recovery of [email protected] was the highest, reaching 61.78 %. Then, the CKMB and [email protected] were characterized by TGA, FT-IR, XRD, BET and SEM, and the results showed that laccase could be well immobilized on CKMB, the maximum enzyme loading could reach 57.5 mg/g. Compared to free laccase, the storage and pH stability of [email protected] was improved greatly. The [email protected] retained about 40 % of relative activity (4 ℃, 30 days) and more than 50 % of relative activity at pH 2-6. In addition, the [email protected] indicated the reusability up to 6 reaction cycles while retaining 45.1 % of relative activity. Moreover, the thermal deactivation kinetic studies of [email protected] showed a lower k value (0.00275 min-1) and higher t1/2 values (252.0 min) than the k value (0.00573 min-1) and t1/2 values (121.0 min) of free laccase.
Conclusions: In summary, we explored scientific and reasonable immobilization conditions of [email protected], and the [email protected] possessed relatively better stabilities, which gave the immobilization of laccase on this cheap and easily available carrier material the possibility of industrial applications.