title: Characterization of a new Acidobacteria-derived moderately thermostable lipase from a Brazilian Atlantic Forest soil metagenome.
abstract: A clone (LP001) expressing a new lipase gene was isolated from a metagenomic library of the Brazilian Atlantic Forest soil. The DNA insert of LP001 was fully sequenced, and 38 ORFs were identified. Comparison of ORFs, %G + C content and gene organization with sequenced bacterial genomes suggested that the fosmid DNA insert belongs to an organism of the Acidobacteria phylum. Protein domain analysis and inactivation by transposon insertion showed that the protein encoded by ORF29 was responsible for the lipase activity and was named LipAAc. The purified LipAAc lipase was capable of hydrolyzing a broad range of substrates, showing the highest activity against p-nitrophenol (pNP) decanoate. The lipase was active over a pH range of 5.0-10.0 and was insensitive to divalent cations. LipAAc is moderately thermostable with optimum temperature between 50 and 60 °C and was thermally activated (80% activity increase) after 1 h incubation at 50 °C. Phylogenetic analysis suggested that the LipAAc is a member of family I of bacterial lipases and clusters with other moderately thermostable lipases of this group. Comparisons of the DNA insert of fosmid LP001 with other acidobacterial genomes and sequence database suggest that lipAAc gene has a fungal origin and was acquired by horizontal transfer.
authors.affiliation: Department of Biochemistry and Molecular Biology, Universidade Federal do Paraná, Curitiba, PR, Brazil.
authors.forename: Helisson
authors.initials: H
authors.lastname: Faoro
authors.forename: Arnaldo
authors.initials: A
authors.lastname: Glogauer
authors.forename: Gustavo Henrique
authors.initials: GH
authors.lastname: Couto
authors.forename: Emanuel Maltempi
authors.initials: EM
authors.lastname: de Souza
authors.forename: Liu Un
authors.initials: LU
authors.lastname: Rigo
authors.forename: Leonardo Magalhães
authors.initials: LM
authors.lastname: Cruz
authors.forename: Rose Adele
authors.initials: RA
authors.lastname: Monteiro
authors.forename: Fábio de Oliveira
authors.initials: Fde O
authors.lastname: Pedrosa
mesh_terms: D061271:Acidobacteria / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins; D001482:Base Composition; D001938:Brazil; D004269:DNA, Bacterial / Q000235:genetics; D015723:Gene Library; D016680:Genome, Bacterial; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D054892:Metagenome; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000378:metabolism; D016366:Open Reading Frames; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D014197:Trees / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009596:Nitrophenols; D008049:Lipase; C428386:thermostable lipase; C024836:4-nitrophenol