Journal of Pure and Applied MicrobiologyVol. 11 No. 3

Application of Box-Behnken Design for the Optimization of Culture Conditions for Novel Fibrinolytic Enzyme Production by Bacillus altitudinis S-CSR 0020

K.V. Smitha1, 2 and B.V. Pradeep1

1Department of Microbiology, Karpagam University, (Karpagam Academy of Higher Education), Coimbatore - 641021, Tamil Nadu, India. 2Department of Microbiology, Safi Institute of Advanced Study, Vazhyoor-673633, Malappuram, India.

Received on 20 June 2017 and accepted on 25 August 2017

 

ABSTRACT

A bacterium which produced novel extracellular fibrinolytic enzyme for digesting bovine blood clots was isolated from soil, and identified by 16s rRNA sequencing as Bacillus altitudinis, given strain name was S-CSR 0020 (accession number KT369312). Fibrin proved the best nitrogen source with an enzyme activity of 750 U/mL, followed by casein after incubation at 37 C for 4 days. The cultural conditions were optimised using Response Surface Methodology (RSM) and Box-Behnken Design (BBD). Based on 3D surface plot and contour plots, the optimized temperature, pH and substrate concentrations were 47 C, 10.5 and 4 g/L respectively, resulted in increase in enzyme activity of 306.88 U/mL and specific activity of 780 U/mg which was 2-fold; compared to initial level of 400 U/mg after 2 days of incubation. The crude enzyme has got potent activity and digested human blood clot completely within 1 hr.

Keywords : Bacillus altitudinis S-CSR 0020, bovine blood clot, Box Behnken Design, fibrinolytic enzyme.