Isolation and Biochemical Characterization of Extracellular Microbial Proteases | ||||
| Delta Journal of Science | ||||
| Article 2, Volume 38, Issue 2, December 2017, Page 142-149 PDF (1.08 MB) | ||||
| Document Type: Research and Reference | ||||
| DOI: 10.21608/djs.2017.139439 | ||||
 View on SCiNiTO | ||||
| Authors | ||||
| Maha M. Salem1; Rasha Abu-Khudir2; Nanis Gamal Allam3; Ehab M. M. Ali4 | ||||
| 1Chemistry Department Biochemistry Division Faculty of Science, Tanta University, Tanta 31527, Egypt | ||||
| 2Chemistry Department, Faculty of Science, Tanta University, Tanta 31527, Egypt. | ||||
| 3Botany Department microbiology unit, Faculty of Science, Tanta University, Tanta 31527, Egypt | ||||
| 4Chemistry Department Biochemistry Division Faculty of Science, Tanta University, Tanta 31527, Egypt. | ||||
| Abstract | ||||
| Proteases are a broad family of hydrolytic enzymes with various applications in chemical, cosmetics, and pharmaceutical industries. Owing to their physiological necessity, proteases are found in diverse sources including microorganisms. Our objective study was to search for a high quality and inexpensive source for the production of microbial proteases under different culture and growth conditions. Also, we aimed to characterize microbial proteases. Proteases-producing bacteria were isolated from soil samples collected from a poultry waste site. Soil samples were inoculated in skimmed agar media and 48 h later, colonies producing clear zones were selected as the source of microorganisms producing enzyme. The isolates were used to inoculate liquid media and the clear supernatant was taken as a crude for enzyme preparation. The enzyme was isolated and purified with ammonium sulfate at 60-80% saturation followed by dialysis. Subsequently, characterization of the enzyme fraction with the highest activity was carried out. The results indicated that the isolated enzyme with (60-80%) fractionation of ammonium sulfate exhibited the highest specific activity. In addition, the optimal temperature for enzyme activity was determined at 70ºC at pH values of 0.05M of acetate buffer 3.6 and 0.05M of glycine-NaOH buffer 10.0. Finally, the kinetic parameters (Michaelis–Menten constant, Km and maximal reaction velocity, Vmax) were calculated as 0.11 μmole/ml and 0.5x104 nmole of tyrosine/ml/hour, respectively. In conclusion, our findings provide evidence that the isolated bacteria represent a rich source of thermostable proteases. Indeed, more studies are still required to obtain such proteases in a purified form | ||||
| Keywords | ||||
| Proteases; Microorganisms; Poultry waste; Skimmed agar media | ||||
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