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Alkaline protease has tremendous applications in different industries, and to fulfil industrial and commercial enzyme requirements, overproduction and optimization of production medium are prerequisites. Protease production by Bacillus amyloliquefaciens SP1, isolated from apple tree rhizosphere was studied. Plackett Burmann design and response surface methodology was used to optimize medium for alkaline protease production. Higher levels of protease activity were observed in presence of disaccharides owing to diauxic growth and inorganic nitrogen sources proved to be less favourable. After screening through one-factor-at-a time experiments and Plackett Burmann design, casein, yeast extract, maltose and KH2PO4 were identified as the most significant variables. Statistical design based three-dimensional (3-D) and contour plots were generated to understand the relationship between enzyme activity and medium variables. The protease production was found to increase from 1730 to 3630 µg/ml/min in optimized medium i.e. approx. 2.1 fold increased compared with original medium. The study assumes significance in the ability of bacterium to survive in wide variety of salts and agricultural wastes and yield optimum levels of extracellular protease. [ABSTRACT FROM AUTHOR]

Alkaline protease from Bacillus amyloliquefaciens SP1 has been characterized in detail for its ecofriendly application of release of silver particles from gelatin layers of used X-ray films. It exhibited optimum activity at broad temperature range and maximum at 60°C under alkaline pH environment (8-12). Thermal inactivation of the crude enzyme followed first order kinetics. The half-life of the enzyme at 50, 60 and 65°C was 70, 15 and 12.6 min, respectively and the denaturation energy was 114.87 kJ/mol. Enzyme retained 53.83 and 108.33% of its initial activity after heating for 15 min at pH 8.0 and temperature 60°C, in presence and absence of 10 mM MnSO4, respectively. Enzymatic decomposition of gelatin layers was enhanced by increase of enzyme concentration from 38 to 3630 µg/ml/min, at 60°C and pH 8.0. This study reported the shortest time of 1.30 min at 3630 µg/ml/min and 4 : 30 min at 74 µg/ml/min of enzyme concentration for hydrolysis of gelatin layers. Keeping in mind that, nowadays recycling is needed and imperative, this is the first study to report that after addition of Mn2+ ions, thermal stability of enzyme increased and it could be effectively reused for 8 cycles as compared with enzyme without protective agents which also increase its yield of silver recovery i.e. 19.56 ± 0.78% by eight times. [ABSTRACT FROM AUTHOR]