OPTIMISATION OF BREVIBACTERIUM, BACILLUS AND PSEUDOMONAS SPECIES FOR PRODUCTION OF LIPASE USING TOBACCO SEED OIL AND OLIVE OIL AS SOLE CARBON SOURCES

Abstract

This study considered the optimization of cultural conditions for enhanced lipolytic enzyme activities amongst 4 bacterial Isolates. (Soil samples from different oil-impact soil areas in Ogun -State Nigeria were obtained with soil auger and transported the Biotechnology Laboratory of Federal Institute of Industrial Research Oshodi, Lagos-Nigeria. Soil was subjected to Microbiology studies such as Primary Isolation. Pure cultures obtained test using tributyrin agar, Lipase - hyper producing bacterial Isolates were identified using biochemical tests and Molecular biology tools (Polymerase chain reaction and gene sequencing) out of a total of 11 bacterial Isolates, 4 bacterial Isolates identified as hyper-pro-ducers were Isolate H (Bavibacillus brevis strain HK 544), Isolate A (Pseudommas aeruinosa strain WES 2), Isolate B (Bcillus megaterium strain WH 13), and Isolate F (Bacillus subtilis BS 01) changes in PH of culture/production medium showed that pH 7.0, favoured the Lipase production in Bacillus subtilis (Isolate F) and Pseudomonas aeruginosa (Isolate A) while pH values of 8.0 favoured Lipase production in Isolate H (Brevibacillus brevis). Variation in temperature showed that the 4 bacterial strain responded best to Mesophilic PH ranges from 25 oC- 40 oC. The optimal Nitrogen source as identified in the study was KNO3 and NH4 No3 at higher concentrations (6 g/C). In comparison, Tobacco seed oil (TSO) competed favorably as carbon source with Olive oil as activities of Lipase from the 4 bacterial Isolates ranged from 8.0 - 28.0 units. The use of lesser known oilbased carbon source like Tobacco seed oil has shown a lot of value addition in terms of commercial value.