Bioprospecting Hot Spring Metagenome: Lipase for the Production of Biodiesel
Screening of metagenomic library from Taptapani Hot Spring (Odisha) yielded a positive lipase clone (pUC-lip479). Sequence analysis showed an ORF (RK-lip479) of 416 amino acid residues which was overexpressed in Escherichia coli BL21 (DE3). Optimum pH and temperature of purified lipase RK-lip479 were 8.0 and 65 °C, respectively, and found to be stable over a pH range of 7.0–9.0 and temperatures 55–75 °C.
RK-lip479 could hydrolyse a wide range of 4-nitrophenyl esters (4-nitrophenyoctanoate, 4-nitrophenyldodecanoate, 4-nitrophenylpalmitate, 4-nitrophenylmyristate and 4-nitrophenylstearate), and maximum activity was observed with 4-nitrophenyldodecanoate. RK-lip479 was resistant to many organic solvents, especially isopropanol, DMSO, methanol, DMF, ethanol, dichloromethane, acetone, glycerol and ethyl acetate.
RK-lip479 also showed activity in the presence of monovalent (Na+ and K+), divalent (Mg2+, Mn2+, Ca2+, Hg2+, Cu2+, Co2+, Zn2+ and Ag2+ ) and trivalent cations (Fe3+ and Al3+). Yield of biodiesel production was in the range of 40–76% using various waste oils with RK-Lip479 under optimized conditions.
Sahoo, R.K., Kumar, M., Sukla, L.B. et al. Environ Sci Pollut Res (2016). doi:10.1007/s11356-016-8118-7