title: Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast.
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.
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR, Brasil.
authors.forename: G H
authors.initials: GH
authors.lastname: Couto
authors.forename: A
authors.initials: A
authors.lastname: Glogauer
authors.forename: H
authors.initials: H
authors.lastname: Faoro
authors.forename: L S
authors.initials: LS
authors.lastname: Chubatsu
authors.forename: E M
authors.initials: EM
authors.lastname: Souza
authors.forename: F O
authors.initials: FO
authors.lastname: Pedrosa
mesh_terms: D001938:Brazil; D004247:DNA / Q000302:isolation & purification; D057075:Enzyme Assays; D015723:Gene Library; D019015:Geologic Sediments / Q000737:chemistry*; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D008066:Lipolysis; D056186:Metagenomics / Q000379:methods*; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D043982:Rhizophoraceae; D012623:Seawater; D017386:Sequence Homology, Amino Acid
chemical_list: D004247:DNA; D008049:Lipase