One-pot synthesis and biochemical characterization of protease metal organic framework (protease@MOF) and its application on the hydrolysis of fish protein-waste

Abstract A protease from Bacillus sp. CHA410 was purified and immobilized by a one-step MOF-embedded approach. The immobilized protease characterized using transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). The optimal pH activity of protease CHA410 and protease@MOF was obtained at pH 8.0 and 9.0, respectively. Thermostability results after 160 min incubation showed that a 25 and 41 % enhancement in the relative activity of protease@MOF observed at 60 and 70 °C, respectively, compared to free protease. Km of the free and immobilized protease@MOF in the presence of casein was 0.685 and 0.033 mg/ml, respectively. Also, Km of the free and immobilized protease@MOF in the presence of fibrin was 0.292 and 0.145 mg/ml, respectively. The Fibrinolytic activity/Caseinolytic activity ratio (F/C ratio) of the free and immobilized proteases was 0.36 and 0.43, respectively. Protease activity of both forms of the enzyme was increased in the presence of some divalent cations, including Ca2+, Mn2+, Mg2+, and Zn2+ ions, while it intensely diminished by phenylmethylsulfonyl fluoride (PMSF), proposed as serine-protease. A 10 and 82 % enhancement in protease activity of free and immobilized proteases was achieved in the presence of butanol, respectively. Storage stability results showed that the immobilized enzyme retained about 70% of its original activity at the end of this period, while the free enzyme only showed 22 % of its initial activity. The hydrolysis degree of immobilized protease CHA410 in the hydrolysis of fish protein waste was obtained about 46% after 2 h of incubation at 50 °C. In comparison, it was gained about 20 % for protease CHA410 at a similar situation. Finally, results indicated that the free and immobilized protease could be used in the food industry for the hydrolysis of fish protein waste.