Research Article

Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast

Published: March 23, 2010
Genet. Mol. Res. 9 (1) : 514-523 DOI: https://doi.org/10.4238/vol9-1gmr738
Cite this Article:
G.H. Couto, A. Glogauer, H. Faoro, L.S. Chubatsu, E.M. Souza, F.O. Pedrosa (2010). Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast. Genet. Mol. Res. 9(1): 514-523. https://doi.org/10.4238/vol9-1gmr738
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Abstract

A novel gene coding for a LipA-like lipase with 283 amino acids and a molecular mass of 32 kDa was isolated and characterized from a metagenomic library prepared from mangrove sediment from the south Brazilian coast. LipA was 52% identical to a lipolytic enzyme from an uncultured bacterium and shared only low identities (≤31%) with lipases/esterases from cultivable microorganisms. Phylogenetic analysis showed that LipA, together with an orthologous protein from an uncultured bacterium, forms a unique branch within family I of true lipases, thereby constituting a new lipase subfamily. Activity determination using crude extracts of Escherichia coli bearing the lipA gene revealed that this new enzyme has a preference for esters with short-chain fatty acids (C ≤ 10) and has maximum activity against p-nitrophenyl-caprate (chain length C10, 0.87 U/mg protein). The optimum pH of LipA was 8.0, and the enzyme was active over a temperature range of 20 to 35°C, with optimum activity against p-nitrophenyl-butyrate at 35°C and pH 8.0.

A novel gene coding for a LipA-like lipase with 283 amino acids and a molecular mass of 32 kDa was isolated and characterized from a metagenomic library prepared from mangrove sediment from the south Brazilian coast. LipA was 52% identical to a lipolytic enzyme from an uncultured bacterium and shared only low identities (≤31%) with lipases/esterases from cultivable microorganisms. Phylogenetic analysis showed that LipA, together with an orthologous protein from an uncultured bacterium, forms a unique branch within family I of true lipases, thereby constituting a new lipase subfamily. Activity determination using crude extracts of Escherichia coli bearing the lipA gene revealed that this new enzyme has a preference for esters with short-chain fatty acids (C ≤ 10) and has maximum activity against p-nitrophenyl-caprate (chain length C10, 0.87 U/mg protein). The optimum pH of LipA was 8.0, and the enzyme was active over a temperature range of 20 to 35°C, with optimum activity against p-nitrophenyl-butyrate at 35°C and pH 8.0.