The Toluene o-Xylene Monooxygenase Enzymatic Activity for the Biosynthesis of Aromatic Antioxidants.
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.
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Giuliana
G
Donadio
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Carmen
C
Sarcinelli
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Elio
E
Pizzo
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Eugenio
E
Notomista
Department of Chemical Sciences, University of Naples Federico II, via Cinthia I, 80126, Napoli, Italy.
Alessandro
A
Pezzella
Department of Sciences and Technologies, University of Sannio, Via Port'Arsa, 11, Benevento, Italy.
Carlo
C
Di Cristo
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Federica
F
De Lise
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy.
Alberto
A
Di Donato
Department of Biology, University of Naples Federico II, Via Cinthia I, 80126, Napoli, Italy; Department of Medicine and Surgery, University of Salerno, via S. Allende, 84081, Baronissi (SA), Italy.
Viviana
V
Izzo
D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002384:Catalysis; D020134:Catalytic Domain; D002460:Cell Line; D004789:Enzyme Activation; D005026:Ethylene Glycols / Q000737:chemistry; D006801:Humans; D006900:Hydroxylation; D007189:Indans / Q000737:chemistry; D010105:Oxygenases / Q000737:chemistry / Q000378:metabolism*; D010797:Phthalimides / Q000737:chemistry; D013379:Substrate Specificity
D000975:Antioxidants; D005026:Ethylene Glycols; D007189:Indans; D010797:Phthalimides; C521491:2-indanol; D010105:Oxygenases; C114360:toluene 2-xylene monooxygenase; C005398:phenoxyethanol; C005020:folpet