Inhibition of inflammatory response in LPS-induced macrophages by 9-KOTE and 13-KOTE produced by biotransformation

Enzyme Microb Technol. 2014 May 10:58-59:36-43. doi: 10.1016/j.enzmictec.2014.02.011. Epub 2014 Mar 5.

Abstract

Lipid mediators such as the leukotrienes, resolvins and protectins have been considered excellent models for the development of new anti-inflammatory drugs, due to their high potentiality. Nevertheless, only tiny amounts are available from natural sources and they have to be prepared by total synthesis. It is known that besides chemical reagents, microorganisms can also promote fatty acid oxygenation, via enzymatic reactions. In this context, the aim of this work was to produce oxylipids analogues in structure to lipid mediators employing microbial biotransformation. To this end, α-linolenic acid (ALA) was biotransformed by the fungi Aspergillus niger into oxylipids with different levels of oxygenation within 24h or 48h. The anti-inflammatory potential of products were evaluated by means of NO and TNF-α quantification in LPS-stimulated RAW264.7 macrophage cell line which guided the isolation of the regioisomers at m/z [M-H](-) 291, 9-keto-10E,12Z,15Z-octadecatrienoic acid (9-KOTE) and 13-keto-9Z,11E,15Z-octadecatrienoic acid (13-KOTE). We showed that biotransformation represents a powerful strategy for the production of potentially interesting candidates for development of anti-inflammation therapies.

Keywords: Biotransformation; Inflammation; Lipid mediators; Liquid chromatography-tandem mass spectrometry; Polyunsaturated fatty acid (PUFA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Aspergillus niger / metabolism*
  • Biocatalysis
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Drug Design
  • Fatty Acids, Unsaturated / isolation & purification
  • Fatty Acids, Unsaturated / metabolism
  • Fatty Acids, Unsaturated / pharmacology*
  • Lipopolysaccharides / pharmacology*
  • Macrophage Activation / drug effects*
  • Macrophages, Peritoneal / drug effects
  • Macrophages, Peritoneal / metabolism
  • Mice
  • Molecular Structure
  • Nitric Oxide / biosynthesis
  • Oxylipins / isolation & purification
  • Oxylipins / metabolism
  • Oxylipins / pharmacology*
  • Tandem Mass Spectrometry
  • Tumor Necrosis Factor-alpha / biosynthesis
  • alpha-Linolenic Acid / metabolism

Substances

  • 9-keto-10(E),12(Z),15(Z)-octadecatrienoic acid
  • Anti-Inflammatory Agents, Non-Steroidal
  • Fatty Acids, Unsaturated
  • Lipopolysaccharides
  • Oxylipins
  • Tumor Necrosis Factor-alpha
  • alpha-Linolenic Acid
  • Nitric Oxide