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Aromatic hydroxylation catalyzed by toluene 4-monooxygenase in organic solvent/aqueous buffer mixtures

Abstract

Toluene 4-monooxygenase is a four-protein component diiron enzyme complex. The enzyme catalyzes the hydroxylation of toluene to give p-cresol with ∼96% regioselectivity. The performance of the enzyme in two-phase reaction systems consisting of toluene, hexane, or perfluorohexane and an aqueous buffer was tested. In each of the cosolvent systems, containing up to 93% (v/v) of solvent, the enzyme was active and exhibited regioselectivity indistinguishable from the aqueous reaction. Using the perfluorohexane/buffer system, a number of polycyclic aromatic hydrocarbons were oxidized that were not readily oxidized in aqueous buffer. An instability of the hydroxylase component and a substantial uncoupling of NADH utilization and product formation were observed in reactions that were continued for longer than ∼3 min. More stable enzyme complexes will be needed for broad applicability of this hydroxylating system in nonaqueous media.

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Correspondence to Brian G. Fox or Jonathan S. Dordick.

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Oppenheim, S.F., Studts, J.M., Fox, B.G. et al. Aromatic hydroxylation catalyzed by toluene 4-monooxygenase in organic solvent/aqueous buffer mixtures. Appl Biochem Biotechnol 90, 187–197 (2001). https://doi.org/10.1385/ABAB:90:3:187

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Index Entries

  • Diiron enzyme
  • monooxygenase
  • organic cosolvents
  • regioselective hydroxylation