Isolation of an organic solvent-tolerant bacterium Bacillus licheniformis PAL05 that is able to secrete solvent-stable lipase

In this study, seven lipase-producing bacterial strains were isolated from salt-enriched and cattle farm soil samples after incubation in toluene- and benzene-enriched media. One strain (PAL05) showed significantly greater lipase activity on spirit blue agar medium and stability in organic solvents. The positive strain (PAL05) was identified as Bacillus licheniformis by 16S rRNA gene sequencing. Lipase production was optimized in a medium containing glycerol as the carbon source and Tween 80 as an inducer (0.5% glycerol + 0.5% Tween 80) at pH 8.0 and a temperature of 30 degrees C. In addition, the enzyme was moderately halotolerant as it exhibited increased activity in the presence of 2.5% NaCl. Optimized conditions increased the lipase production threefold. Crude lipase retained its activity for 14 days of incubation in the presence of various organic solvents at a level of 25% and 50%. The enzyme was stable at 25% in most solvents; some of the solvents such as hexane, benzene, and ethanol actually stimulated enzyme activity. The organic solvent stability of the lipase produced by the strain PAL05 enables the enzyme to be used as a potential biocatalyst for ester synthesis and other applications in nonaqueous conditions. (C) 2014 International Union of Biochemistry and Molecular Biology, Inc.