Gene cloning and characterization of a novel highly organic solvent tolerant lipase from Proteus sp. SW1 and its application for biodiesel production

Mol Biotechnol. 2013 Jan;53(1):55-62. doi: 10.1007/s12033-012-9518-7.

Abstract

Proteus sp. SW1 was found to produce an extracellular solvent tolerant lipase. The gene, lipA, encoding a bacterial lipase, was cloned from total Proteus sp. SW1 DNA. lipA was predicted to encode a 287 amino acid protein of 31.2 kDa belonging to the Group I proteobacterial lipases. Purified His-tagged LipA exhibited optimal activity at pH 10.0 and 55°C. It was highly stable in organic solvents retaining 112% of its activity in 100% isopropanol after 24 h, and exhibited more than 200% of its initial activity upon exposure to 60% acetone, ethanol, and hexane for 18 h. Biodiesel synthesis reactions, using a single step addition of 13% an acyl acceptor ethanol, showed that LipA was highly effective at converting palm oil into biodiesel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biofuels / microbiology*
  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • Cloning, Molecular*
  • Esterification
  • Gas Chromatography-Mass Spectrometry
  • Gene Library
  • Hexanes / metabolism
  • Hydrogen-Ion Concentration
  • Lipase / biosynthesis*
  • Lipase / genetics
  • Palm Oil
  • Plant Oils / metabolism
  • Plasmids
  • Proteus / enzymology*
  • Proteus / genetics
  • RNA, Ribosomal, 16S
  • Sequence Analysis, RNA
  • Solvents / chemistry
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Biofuels
  • Hexanes
  • Plant Oils
  • RNA, Ribosomal, 16S
  • Solvents
  • n-hexane
  • Palm Oil
  • Lipase