The Toluene o-Xylene Monooxygenase Enzymatic Activity for the Biosynthesis of Aromatic Antioxidants

PLoS One. 2015 Apr 27;10(4):e0124427. doi: 10.1371/journal.pone.0124427. eCollection 2015.

Abstract

Monocyclic phenols and catechols are important antioxidant compounds for the food and pharmaceutic industries; their production through biotransformation of low-added value starting compounds is of major biotechnological interest. The toluene o-xylene monooxygenase (ToMO) from Pseudomonas sp. OX1 is a bacterial multicomponent monooxygenase (BMM) that is able to hydroxylate a wide array of aromatic compounds and has already proven to be a versatile biochemical tool to produce mono- and dihydroxylated derivatives of aromatic compounds. The molecular determinants of its regioselectivity and substrate specificity have been thoroughly investigated, and a computational strategy has been developed which allows designing mutants able to hydroxylate non-natural substrates of this enzyme to obtain high-added value compounds of commercial interest. In this work, we have investigated the use of recombinant ToMO, expressed in cells of Escherichia coli strain JM109, for the biotransformation of non-natural substrates of this enzyme such as 2-phenoxyethanol, phthalan and 2-indanol to produce six hydroxylated derivatives. The hydroxylated products obtained were identified, isolated and their antioxidant potential was assessed both in vitro, using the DPPH assay, and on the rat cardiomyoblast cell line H9c2. Incubation of H9c2 cells with the hydroxylated compounds obtained from ToMO-catalyzed biotransformation induced a differential protective effect towards a mild oxidative stress induced by the presence of sodium arsenite. The results obtained confirm once again the versatility of the ToMO system for oxyfunctionalization reactions of biotechnological importance. Moreover, the hydroxylated derivatives obtained possess an interesting antioxidant potential that encourages the use of the enzyme for further functionalization reactions and their possible use as scaffolds to design novel bioactive molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / chemistry
  • Antioxidants / metabolism*
  • Antioxidants / pharmacology
  • Catalysis
  • Catalytic Domain
  • Cell Line
  • Enzyme Activation
  • Ethylene Glycols / chemistry
  • Humans
  • Hydroxylation
  • Indans / chemistry
  • Oxygenases / chemistry
  • Oxygenases / metabolism*
  • Phthalimides / chemistry
  • Substrate Specificity

Substances

  • Antioxidants
  • Ethylene Glycols
  • Indans
  • Phthalimides
  • 2-indanol
  • Oxygenases
  • toluene 2-xylene monooxygenase
  • phenoxyethanol
  • folpet

Grants and funding

This research was supported by FARB 2013 ORSA 132082. Dr. C. Sarcinelli was supported by grant POR CAMPANIA FSE 2007/2013, Progetto CARINA CUP B25B0900080007. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.