Nonenzymatic Regeneration of Styrene Monooxygenase for Catalysis

Paul, Caroline E.; Tischler, Dirk; Riedel, Anika; Heine, Thomas; Itoh, Nobuhide; Hollmann, Frank

doi:10.1021/acscatal.5b00041

Cited by 73 publications

(93 citation statements)

References 36 publications

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“…Active GEMs require for regeneration of reduced FAD . Under cell‐free conditions, we used either a reductase ( Pf StyB, Ro StyBart) or 1‐benzyl‐1,4‐dihydronicotinamide (BNAH) as stoichiometric reductant, respectively . The reaction conditions were applied as described previously .…”

Section: Resultsmentioning

confidence: 99%

Accessing Enantiopure Epoxides and Sulfoxides: Related Flavin‐Dependent Monooxygenases Provide Reversed Enantioselectivity

et al. 2019

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Enantiopure organic compounds are of major importance for the chemical and pharmaceutical industry. Flavin‐dependent group E monooxygenases, composed of monooxygenase and reductase, are known to perform epoxidation of substituted alkenes as well as sulfoxidation in a regio‐ and enantioselective fashion. Group E is divided into styrene monooxygenases (SMO) and indole monooxygenases (IMO). Hitherto mainly SMOs have been characterized. In this study, we assayed 31 monooxygenases from both types, while 23 of which showed activity. They almost exclusively produced (S)‐styrene oxide at high enantiomeric excess with maximum activities of 0.73 μmol min−1 mg−1 (kcat=0.54 s−1). In case of sulfoxidation, we found that the enantioselectivity is contrary between both types. IMOs preferably produce the (S)‐enantiomer while SMOs have a tendency to produce the (R)‐enantiomer. Sequence analysis and molecular docking of substrates allowed identifying fingerprint motives: SMO N46‐V48‐H50‐Y73‐H76‐S96 and IMO S46‐Q48‐M50‐V/I73‐I76‐A96. These form an essential part of the active site while the loop (AS44‐51) interacts with the co‐substrate and other amino acids direct the substrate. The motives clearly distinguish group E monooxygenases and define the enantioselectivity and thus direct biotechnological applications. Two‐hour biotransformations with several sulfides in conjunction with upscale experiments (10 and 100 mg scale) resulted in the identification of promising candidates for the realization of biocatalytic processes.

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Section: Resultsmentioning

confidence: 99%

Accessing Enantiopure Epoxides and Sulfoxides: Related Flavin‐Dependent Monooxygenases Provide Reversed Enantioselectivity

et al. 2019

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“…Die prinzipielle Machbarkeit eines solchen stark vereinfachten Reaktionsschemas konnte kürz-lich gezeigt werden (Abb. 3, [7]). …”

Section: Synthetische Nikotinamide In Der Biokatalyseunclassified

“…B. mittels Formiatdehydrogenase(FDH)-katalysierter Oxidation von Ameisensäure. Unten: der vereinfachte FADH 2 -Regenerationszyklus mit mNADH [7].…”

Section: Abbunclassified

Synthetische Nikotinamide in der Biokatalyse

Hollmann

Paul

2015

Biospektrum

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NIEDERLANDENatural nicotinamide cofactors (NAD(P)) can in some cases be replaced by simple synthetic analogues (mNADs). These analogues can enable 'faster than naturally designed' enzymatic reactions, more efficient reaction processes and more chemoselective reactions because of their bioorthogonality.ó Die Nikotinamidkofaktoren Nikotinamidadenindinukleotid (NAD) und Nikotinamidadenindinukleotidphosphat (NADP) sind die zentralen Redoxkoenzyme aller bekannten Lebewesen. Da sie für viele enzymatische Redoxreaktionen unabdingbar sind, spielen sie auch in der Biokatalyse eine wichtige Rolle. In den letzten Jahrzehnten lag ein Schwerpunkt in der Redoxbiokatalyse-Forschung auf der Entwicklung effizienter in situ-Regenerationssysteme, die einen katalytischen Einsatz von NAD(P) ermöglichen.

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“…We previously showed the reduction rate of the flavin cofactor varies according to the NCB substituents and redox potential in ene reductases . Additionally, NCBs directly reduce free flavins in solution . Here we investigate the use of NCBs with group A flavoprotein hydroxylases.…”

Section: Introductionmentioning

confidence: 96%

Flavoenzyme‐mediated Regioselective Aromatic Hydroxylation with Coenzyme Biomimetics

et al. 2020

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Regioselective aromatic hydroxylation is desirable for the production of valuable compounds. External flavin‐containing monooxygenases activate and selectively incorporate an oxygen atom in phenolic compounds through flavin reduction by the nicotinamide adenine dinucleotide coenzyme, and subsequent reaction with molecular oxygen. This study provides the proof of principle of flavoenzyme‐catalyzed selective aromatic hydroxylation with coenzyme biomimetics. The carbamoylmethyl‐substituted biomimetic in particular affords full conversion in less than two hours for the selective hydroxylation of 5 mM 3‐ and 4‐hydroxybenzoates, displaying similar rates as with NADH, achieving a 10 mM/h enzymatic conversion of the medicinal product gentisate. This biomimetic appears to generate less uncoupling of hydroxylation that typically leads to undesired hydrogen peroxide. Therefore, we show these flavoenzymes have the potential to be applied in combination with biomimetics.

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Nonenzymatic Regeneration of Styrene Monooxygenase for Catalysis

Cited by 73 publications

References 36 publications

Accessing Enantiopure Epoxides and Sulfoxides: Related Flavin‐Dependent Monooxygenases Provide Reversed Enantioselectivity

Accessing Enantiopure Epoxides and Sulfoxides: Related Flavin‐Dependent Monooxygenases Provide Reversed Enantioselectivity

Synthetische Nikotinamide in der Biokatalyse

Flavoenzyme‐mediated Regioselective Aromatic Hydroxylation with Coenzyme Biomimetics

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