Usage of GD-95 and GD-66 lipases as fusion partners leading to improved chimeric enzyme LipGD95-GD66

Int J Biol Macromol. 2018 Oct 15;118(Pt B):1594-1603. doi: 10.1016/j.ijbiomac.2018.07.002. Epub 2018 Jul 5.

Abstract

Lipases are used as biocatalysts in industrial processes mainly because of their stability at broad temperature and pH range, resistance to organic solvents and wide spectrum of substrates. The usage of several lipolytic domains, each with different activity and resistance profiles, enables both the flexibility and efficiency of industrial processes. In this study, GD-95 and GD-66 lipases produced by Geobacillus sp. 95 and Geobacillus sp. 66, respectively, were used as fusion partners to create a new fused lipolytic enzyme LipGD95-GD66. Chimeric LipGD95-GD66 lipase displayed tenfold increase in activity (200 U/mg) compared to parental GD-66 lipase, improved Vmax (10 μmol/min mg-1) and catalytic efficiency (2 ∗ 105 min-1 mM-1) for p-NP palmitate as a substrate and increased activity at 70-75 °C compared to both parental lipases. All three lipases also retained >50% of their lipolytic activity after incubation with methanol, n-hexane, ethanol and DMF for longer than three weeks, highlighting a great prospect for application in industrial processes. Moreover, transesterification results revealed the capability of parental GD-95 lipase to be the most promising biocatalyst for production of methyl and ethyl esters through eco-friendly transesterification using argan oil and ethanol/methanol as acceptors of acyl group.

Keywords: Ester synthesis; Fused enzymes; Geobacillus lipases.

MeSH terms

  • Biocatalysis
  • Catalytic Domain
  • Enzyme Inhibitors / pharmacology
  • Enzyme Stability
  • Esterification
  • Geobacillus / enzymology
  • Geobacillus / genetics
  • Hydrogen-Ion Concentration
  • Kinetics
  • Lipase / antagonists & inhibitors
  • Lipase / chemistry
  • Lipase / genetics*
  • Lipase / metabolism*
  • Metals / pharmacology
  • Protein Engineering*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism*
  • Solvents / pharmacology

Substances

  • Enzyme Inhibitors
  • Metals
  • Recombinant Fusion Proteins
  • Solvents
  • Lipase