PRODUCTION AND PARTIAL PURIFICATION OF LIPASE FROM BACILLUS OLERONIUS B

AUTHOR: OKAFOR, FRANCIS DOZIE

DEPARTMENT: APPLIED MICROBIOLOGY AND BREWING

AFFILIATION: NNAMDI AZIKIWE UNIVERSITY, AWKA

Four bacterial isolates from palm oil contaminated soil in Awka, Nigeria were screened for lipase production in a submerged culture. The isolate with highest lipase activity was characterized and identified as Bacillus oleronius B based on morphological, biochemical characteristics and 16S rRNA sequencing. Lipase was maximally produced when 10ml of palm oil was used as the carbon source in the growth of Bacillus oleronius B. A mixture of yeast extract and peptone stimulated lipase production in Bacillus oleronius B. Lipase accumulation was also enhanced by K+ in Bacillus oleronius B. Of the pH levels studied, pH 7.0 gave the highest yield of lipase in Bacillus oleronius B. Time course experiment for lipase production in Bacillus oleronius B showed highest lipase yield after 48 hours. The lipase was separated by carboxy methyl (CM) Sepharose, ion exchange chromatography. The optimum temperature for enzyme activity was 60oC. The lipase showed optimum activity at pH 7.0. The Km value for palm oil, olive oil, groundnut oil and palm kernel oil was 0.35%, 0.61%, 0.42% and 0.48%, respectively. Co2+, Hg2+, Zn2+, Cu2+, Mn2+, Li+, Fe2+, Mg2+, EDTA and SDS were inhibitory to the lipase.

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