Structural and functional characterization of a novel lipolytic enzyme from a Brazilian Cerrado soil metagenomic library

Paula Istvan, Amanda Araújo Souza, Aisel Valle Garay, Debora Farage Knupp dos Santos, Gideane Mendes de Oliveira, Renata Henrique Santana, Fabyano Alvares Cardoso Lopes, Sonia Maria de Freitas, João Alexandre Ribeiro Gonçalves Barbosa, Ricardo Henrique Krüger

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Objective: To isolate putative lipase enzymes by screening a Cerrado soil metagenomic library with novel features. Results: Of 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. Conclusion: Our results demonstrate the isolation of novel family V lipolytic enzyme with biotechnological applications from a metagenomic library.

Original languageEnglish
Pages (from-to)1395-1406
Number of pages12
JournalBiotechnology Letters
Volume40
Issue number9-10
DOIs
StatePublished - 1 Oct 2018
Externally publishedYes

Keywords

  • Cerrado
  • Lipolytic enzymes
  • Metagenome
  • Soil

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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