An efficient chitinase from Bacillus amyloliquefaciens M6: Immobilization to ZnO/RGO nanocomposite and its application in biocontrol


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Uçar S., Dikbaş N., Alım Ş., Çepni E., Tozlu G., Şenol Kotan M., ...Daha Fazla

Journal of Stored Products Research, cilt.116, ss.1-17, 2026 (Hakemli Dergi)

Özet

In this study, the chitinase enzyme purified from Bacillus amyloliquefaciens M6 was immobilized on Zinc oxide/

reduced graphene oxide nanocomposites (ZnO/RGO NCs), and its activity on Rhyzopertha dominica and Ory-

zaephilus surinamensis was tested. To optimize the culture medium, the M6 strain was developed under varying

conditions, and the optimum substrate concentration, temperature, pH, and incubation time for chitinase pro-

duction were determined as 5 g/L chitin, 35 ◦C, 6.0 pH, and 24 h. Compared to its activity prior to optimization

(10.42 U/mL), the chitinase activity of the strain was observed to increase by 2.2 times. Under these conditions,

chitinase was purified 6.75-fold from strain M6 with a specific activity of 228.90 U/mg (activity: 11,48 U/mL)

and a yield of 1.84 %, and its molecular weight was determined to be ~50 kDa. Chitinase was immobilized on

ZnO/RGO NCs with 99.68 % immobilization efficiency and 94.30 % recovered activity. The optimum pH and

temperature values of free and immobilized chitinase were determined as 6.0 and 45 ◦C. The activity of

immobilized chitinase against R. dominica and O. surinamensis was tested and the LC50 and LC90 values for

R. dominica were calculated as 1.8 mg/mL (32.8 U/mL) and 3.8 mg/mL (69.2 U/mL) and 4.8 mg/mL (87.4 U/

mL) and 10.0 mg/mL (182.0 U/mL) for O. surinamensis. Thus, the obtained results concluded that the immo-

bilized chitinase proved an alternative biocontrol agent to hazardous synthetic pesticides, the application of this

biocontrol agent to make biopesticides will prove more reliable for sustaining the crop productivity and in crop

protection.