| Institution: | 1.Laboratório de Enzimologia, Departamento de Biologia Celular, Instituto Central de Ciências Sul,Universidade de Brasília – UnB,Brasília,Brazil;2.Laboratório de Biofísica, Departamento de Biologia Celular,Universidade de Brasília,Brasília,Brazil;3.Instituto Federal de Brasília,Brasília,Brazil |
| Abstract: | ObjectiveTo isolate putative lipase enzymes by screening a Cerrado soil metagenomic library with novel features.ResultsOf 6720 clones evaluated, Clone W (10,000 bp) presented lipolytic activity and four predicted coding sequences, one of them LipW. Characterization of a predicted esterase/lipase, LipW, showed 28% sequence identity with an arylesterase from Pseudomonas fluorescens (pdb|3HEA) from protein database (PDB). Phylogenetic analysis showed LipW clustered with family V lipases; however, LipW was clustered in different subclade belonged to family V, suggesting a different subgroup of family V. In addition, LipW presented a difference in family V GH motif, a glycine replaced by a serine in GH motif. Estimated molecular weight and stokes radius values of LipW were 29,338.67–29,411.98 Da and 2.58–2.83 nm, respectively. Optimal enzyme activity was observed at pH 9.0–9.5 and at 40 °C. Circular dichroism analysis estimated secondary structures percentages as approximately 45% α-helix and 15% β-sheet, consistent with the 3D structure predicted by homology.ConclusionOur results demonstrate the isolation of novel family V lipolytic enzyme with biotechnological applications from a metagenomic library. |
