Bioprospecting hot spring metagenome: lipase for the production of biodiesel
2017
Sahoo, Rajesh Kumar | Mohit, Kumar | Sukla, Lala Behari | Subudhi, Enketeswara
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.
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