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

Genet Mol Res. 2010 Mar 23;9(1):514-23. doi: 10.4238/vol9-1gmr738.

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 (< or =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 < or = 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 degrees C, with optimum activity against p-nitrophenyl-butyrate at 35 degrees C and pH 8.0.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brazil
  • DNA / isolation & purification
  • Enzyme Assays
  • Gene Library*
  • Geologic Sediments / chemistry*
  • Lipase / isolation & purification*
  • Lipase / metabolism
  • Lipolysis
  • Metagenomics / methods*
  • Phylogeny
  • Plasmids / genetics
  • Rhizophoraceae*
  • Seawater*
  • Sequence Homology, Amino Acid

Substances

  • DNA
  • Lipase