UNVEILING A THERMO-ALKALIPHILIC BACTERIAL ISOLATE AS POTENTIAL LIPASE PRODUCER FROM LOCAL WATERBODY, KOCHARVA, VAPI, INDIA: MEDIA OPTIMIZATION STUDIES

Abstract

Our study reports thermo-alkaliphilic bacterial isolate C-1, with lipolytic potential, obtained from a local water body, located in Kocahrva village, Vapi, Gujarat, India, loaded with industrial effluents, identified as Bacillus subtilis, based on biochemical and molecular characterization (GenBank Accession no. MN068818). The present study is directed towards media parameters optimization for Bacillus subtilis C-1 lipase productio, using Plackett-Burman Design and response surface methodology. Various media variables viz., glucose, olive oil, MgSO₄, ammonium chloride, KH₂PO₄, peptone, tryptone, and CaCl₂ were tested using 16 runs design. In accordance with the Pareto chart, glucose, peptone, MgSO₄ and CaCl₂ were significant factors. The maximum lipase activity obtained was 21.48 Enzyme Units (EU)/ml. These significant media variables were further optimized for concentration, using randomized Central Composite Design, prepared using Design Expert (v.12.0.1). The lipolytic activity increased to 32.1 EU/ml, which was 1.49 times greater, with 3 gm% glucose, 3.5 gm/L peptone and 0.5 gm/L MgSO₄. Biochemical characterization of partially purified C-1 lipase, demonstrated optimum pH 10.0 and optimum temperature for 55^oC. Hence, it was accomplished that Bacillus subtilis C-1 strain can be a prospective lipase source for large scale production.