Open AccessCharacterization of a new A cidobacteria‐ derived moderately thermostable lipase from a B razilian A tlantic F orest soil metagenome
Author(s) -
Faoro Helisson,
Glogauer Arnaldo,
Couto Gustavo Henrique,
Souza Emanuel Maltempi,
Rigo Liu Un,
Cruz Leonardo Magalhães,
Monteiro Rose Adele,
Oliveira Pedrosa Fábio
Publication year - 2012
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1111/j.1574-6941.2012.01361.x
Subject(s) - fosmid , biology , acidobacteria , lipase , orfs , metagenomics , insert (composites) , genetics , gene , genome , biochemistry , actinobacteria , open reading frame , peptide sequence , enzyme , 16s ribosomal rna , mechanical engineering , engineering
A clone ( LP 001) expressing a new lipase gene was isolated from a metagenomic library of the B razilian A tlantic F orest soil. The DNA insert of LP 001 was fully sequenced, and 38 ORF s were identified. Comparison of ORF s, % G + C content and gene organization with sequenced bacterial genomes suggested that the fosmid DNA insert belongs to an organism of the A cidobacteria phylum. Protein domain analysis and inactivation by transposon insertion showed that the protein encoded by ORF 29 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 (p NP ) decanoate. The lipase was active over a p H 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 LP 001 with other acidobacterial genomes and sequence database suggest that lipAAc gene has a fungal origin and was acquired by horizontal transfer.