Journal
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
Volume 24, Issue 4, Pages 3802-3809Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s11356-016-8118-7
Keywords
Hot spring; Metagenomic; Lipase; Solvent tolerance; Metal tolerance; Biodiesel
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Funding
- Department of Biotechnology, Government of India [BT/PR7944/BCE/8/1036/2013]
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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 degrees C, respectively, and found to be stable over a pH range of 7.0-9.0 and temperatures 55-75 degrees 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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