title: Bioprospecting hot spring metagenome: lipase for the production of biodiesel.
abstract: 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.
authors.affiliation: Centre of Biotechnology, Siksha 'O' Anusandhan University, Kalinga Nagar, Bhubaneswar, Odisha, India.
authors.forename: Rajesh Kumar
authors.initials: RK
authors.lastname: Sahoo
authors.affiliation: Department of Biotechnology, Mahatma Jyoti Rao Phoole University, Jaipur, 303002, India. kumarmohit@yahoo.com.
authors.forename: Mohit
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Multi-disciplinary Research Cell, Siksha 'O' Anusandhan University, Khandagiri Square, Bhubaneswar, 751030, India.
authors.forename: Lala Behari
authors.initials: LB
authors.lastname: Sukla
authors.affiliation: Centre of Biotechnology, Siksha 'O' Anusandhan University, Kalinga Nagar, Bhubaneswar, Odisha, India. enketeswarasubudhi@soauniversity.ac.in.
authors.forename: Enketeswara
authors.initials: E
authors.lastname: Subudhi
mesh_terms: D056804:Biofuels; D065132:Bioprospecting; D045482:Hot Springs; D008049:Lipase / Q000378:metabolism*; D054892:Metagenome; D013696:Temperature
chemical_list: D056804:Biofuels; D008049:Lipase
keywords: Biodiesel; Hot spring; Lipase; Metagenomic; Metal tolerance; Solvent tolerance