Abstract The present work sought to broaden the repertoire of proteolytic biocatalyst formulations by producing, purifying, characterizing, and identifying suitable immobilization carriers for an extracellular protease from the filamentous fungi Periconia byssoides . Enzyme production was carried out through submerged fermentation using two different growth media. The peptone‐containing medium proved to be the most suitable for producing extracellular protease. The crude enzyme was partially purified by 70% ethanol precipitation to enable subsequent biochemical characterization and formulation studies. A Doehlert‐type design of experiments was used to determine the optimal pH and temperature of the enzyme, resulting in a pH of 6.4 and a temperature of 70 °C. To evaluate the influence of salts on enzyme activity, various salts were tested at concentrations of 0.1 and 0.01 m . At 0.1 m , CaCl 2 , MgCl 2 and NH 4 Cl inactivated the enzyme. At 0.01 m , BaCl 2 increased the catalytic activity by 6% whereas CaCl 2 and NH 4 Cl inactivated the enzyme. The kinetic parameters of the enzyme were estimated using the Michaelis–Menten equation, with casein as the substrate, at various concentrations. K m and V max were determined to be 0.23 mg mL −1 and 1.26 U mg −1 protein, respectively. A preliminary screening to identify the most efficient carrier for protease immobilization was performed using eight commercial carriers: silica gel, celite 545, chitosan, MAT 540, CPC 27709, Purolite Lifetech ECR8806F, ECR8204F, and ECR1090F. Among these, MAT 540 exhibited 49.93% immobilization yield and 90% protein retention. Overall, this work not only characterizes an extracellular P. byssoides protease but also identifies a promising carrier for its immobilization, supporting the development of an active and stable biocatalyst for future biotechnological applications.
Oliveira et al. (Thu,) studied this question.