A novel solvent tolerant esterase of GDSGG motif subfamily from solar saltern through metagenomic approach: Recombinant expression and characterization

Int J Biol Macromol. 2018 Nov:119:393-401. doi: 10.1016/j.ijbiomac.2018.06.057. Epub 2018 Jun 13.

Abstract

A novel esterase, designated as EstSP was identified by function based screening from a soil metagenomic fosmid library of solar saltern of Goa. EstSP gene of 1065 bp encoding a putative esterase of 354 amino acids showing 55% identity to esterase from gamma proteobacterium HIMB55 was identified. The enzyme EstSP belongs to family IV hormone sensitive lipase with novel sequence characteristics and a unique motif GDSGG. EstSP expressed as a His-tag fusion protein of mass 58 kDa was visualized on SDS PAGE and confirmed by Western blot analysis. The enzyme is an alkaline esterase that exhibited highest catalytic activity towards p-nitrophenyl acetate with optimum temperature 40 °C and pH 8.0. The catalytic efficiency and specific activity of EstSP for p-nitrophenyl acetate was 7407.4 min-1 mM-1 and 915.23 U mg-1 respectively. EstSP showed remarkable stability in the presence of polar and non-polar solvents, retaining >80% of its activity after 72 h. Furthermore, the enzyme is halotolerant with optimum activity at 1 M NaCl and maintained 60% residual activity after 24 h exposure to 5 M NaCl. This novel enzyme with remarkable properties could be a promising candidate for industrial bioprocesses in non-aqueous media as well as pharmaceutical, food and biotechnological applications.

Keywords: Esterase; Metagenomic library; Organic solvent tolerance; Salt tolerance; Solar saltern.

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Cloning, Molecular
  • Enzyme Inhibitors / pharmacology
  • Esterases / antagonists & inhibitors
  • Esterases / chemistry*
  • Esterases / genetics
  • Esterases / metabolism*
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Metagenomics*
  • Metals / pharmacology
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Salts / pharmacology
  • Sequence Analysis
  • Soil
  • Solvents / pharmacology*
  • Substrate Specificity
  • Temperature

Substances

  • Enzyme Inhibitors
  • Metals
  • Recombinant Proteins
  • Salts
  • Soil
  • Solvents
  • Esterases