Characterization of novel metagenomic–derived lipase from Indian hot spring
Extreme environments are the main source of industrially suitable biocatalysts. The non-cultivable approach of searching enzymes is known to provide ample scope to accomplish novelty for their industrial applications. Lip479 clone out of seven lipase-producing clones obtained from Taptapani hot spring was found to be optimally active at pH 8.0 and temperature 65 °C. The recombinant Lip479 was highly stable in organic solvents, methanol, DMF, DMSO, acetone, and dichloromethane. Lip479 lipase activity was enhanced in the presence of K+, Mn2+, Na+, Zn2+, and Ca2+ except for Fe3+. The ability of Lip479 lipase to act on long carbon chain of 4-nitrophenyl myristate suggests it might be a true lipase. Lip479 clone was found to have ORF of 1251 bp encoding 416 amino acid residues of 42.57 KDa size (theoretically calculated). The presence of conserved motif Ala-His-Ser-Gln-Gly and Zn2+-binding consensus sequence (GAAHAAKH) of the clone assigns the protein to lipase family 1.5. Phylogenetic lineage of the protein sequence of Lip479 was traced to family 1.5 as it was clubbed up with those of reported lipases of the same family. The above biochemical features indicated that Lip479 lipase can be a potential biocatalyst for its use in various industries.
KeywordsHot spring Metagenomic Thermostability Lipase
Present research work received the financial support from the DBT, New Delhi through project grants no: BT/PR7944/BCE/8/1036/2013).
Compliance with ethical standards
Conflict of interest
The authors declare that they do not have any conflict of interest.
- Ahmed EH, Raghavendra T, Madamwar D (2010) A thermostable alkaline lipase from a local isolate Bacillus subtilis EH 37: characterization, partial purification, and application in organic synthesis. Appl Biochem Biotechnol 160:2102–2113. https://doi.org/10.1007/s12010-009-8751-4 CrossRefGoogle Scholar
- Aldercreutz P, Mattiasson B (1987) Aspects of biocatalyst stability in organic solvents. Biocatal Biotransform 1:99–108Google Scholar
- Glogauer A, Martini VP, Faoro H, Couto GH, Müller-Santos M, Monteiro RA, Mitchell DA, de Souza EM, Pedrosa FO, Krieger N (2011) Identification and characterization of a new true lipase isolated through metagenomic approach. Microb Cell Factories 10:54. https://doi.org/10.1186/1475-2859-10-54 CrossRefGoogle Scholar
- Jinwal UK, Roy U, Chowdhury AR, Bhaduri AP, Roy PK (2003) Purification and characterization of an alkaline lipase from a newly isolated Pseudomonas mendocina PK-12CS and chemoselective hydrolysis of the fatty acid ester. Bioorg Med Chem 11:1041–1046. https://doi.org/10.1016/S0968-0896(02)00516-3 CrossRefGoogle Scholar
- Sahoo RK (2016) Pure culture and metagenomic approaches to investigate Taptapani hot spring for lipase gene, Dissertation, Siksha O Anusnadhan University, IndiaGoogle Scholar
- Selvin J, Kennedy J, Lejon DPH, Kiran G, Dobson ADW (2012) Isolation identification and biochemical characterization of a novel halo-tolerant lipase from the metagenome of the marine sponge Haliclona simulans. Microb Cell Factories 11:72. https://doi.org/10.1186/1475-2859-11-72 CrossRefGoogle Scholar