[01] Temam Abrar Hamza, Department of Biotechnology, College of Natural Sciences, Arba Minch University, Arba Minch, Ethiopia. [02] Fantahun Woldesenbet, Department of Biotechnology, College of Natural Sciences, Arba Minch University, Arba Minch, Ethiopia.

Proteases are multipurpose group of enzymes used in various industries such as detergent, silver recovery, food, pharmaceutical, leather, and textile industries. This work aimed to produce protease from local microflora for use as detergent additive and silver recovery from waste X-ray film. Isolation of protease producer undertaken using skim milk agar medium and process parameters was optimized. Crude enzyme was characterized followed by stain and gelatine removal tests. A total of 22 protease positive bacteria were isolated from the study area. Out of 22 protease producing bacteria, one isolate designated as Bacillus sp. CAMA14 was selected. Maximum protease production was achieved at 2.5% (v/v) inoculums size, 1% (w/v) NaCl, pH 8.0 at 30°C and 48 hr. Glucose and yeast extract were best carbon and nitrogen sources respectively. The optimum activity was reached at pH 9 and 50°C. The enzyme was stable in the pH range of 7 to 10. It retained 75%, 86% and 72% activity after one hr pre-incubation at 50°C, in 15% H₂O₂ and 0.3% commercial detergent respectively. The enzyme activity was increased by Mg²⁺, Cu²⁺, and Mn²⁺, was not affected by Ca²⁺ but decreased by Zn²⁺, Hg²⁺ and Fe²⁺. It removed stains of egg yolk on cotton cloth, and gelatin on X-ray film at pH 9, 40°C, in 40-46 min. These properties suggest that protease from Bacillus sp. CAMA14 could find potential application in detergent industries and silver recovery process.

Bacillus Sp. CAMA14, Detergent, Gelatine, Protease, X-ray Films

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