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Biocatalytic Aromaticity-Breaking Epoxidation of Naphthalene and Nucleophilic Ring-Opening Reactions

Zhang, W. (author), Li, Huanhuan (author), Younes, S.H.H. (author), Gómez De Santos, Patricia (author), Tieves, F. (author), Grogan, Gideon (author), Pabst, Martin (author), Alcalde, Miguel (author), Whitwood, Adrian C. (author), Hollmann, F. (author)

Aromatic hydroxylation reactions catalyzed by heme-thiolate enzymes proceed via an epoxide intermediate. These aromatic epoxides could be valuable building blocks for organic synthesis giving access to a range of chiral trans-disubstituted cyclohexadiene synthons. Here, we show that naphthalene epoxides generated by fungal peroxygenases can...

journal article 2021

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Pilot-scale production of peroxygenase from agrocybe aegerita

Tonin, F. (author), Tieves, F. (author), Willot, S.J. (author), Van Troost, Anouska (author), van Oosten, R.J.C. (author), Breestraat, Stefaan (author), Van Pelt, Sander (author), Alcalde, Miguel (author), Hollmann, F. (author)

The pilot-scale production of the peroxygenase from Agrocybe aegerita (rAaeUPO) is demonstrated. In a fed-batch fermentation of the recombinant Pichia pastoris, the enzyme was secreted into the culture medium to a final concentration of 0.29 g L-1 corresponding to 735 g of the peroxygenase in 2500 L of the fermentation broth after 6 days. Due...

journal article 2021

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Selective Oxyfunctionalisation Reactions Driven by Sulfite Oxidase-Catalysed In Situ Generation of H₂O₂

van Schie, M.M.C.H. (author), Kaczmarek, Alexander T. (author), Tieves, F. (author), Gomez de Santos, Patricia (author), Paul, C.E. (author), Arends, I.W.C.E. (author), Alcalde, Miguel (author), Schwarz, Günter (author), Hollmann, F. (author)

H₂O₂ can be accepted by several peroxygenases as a clean oxidant, able to supply both the necessary electrons and oxygen atom at the same time. The biocatalysts, in turn, are able to catalyse an array of interesting oxygen insertion reactions at enantio- and regio-selectivities hard to attain with classical chemical...

journal article 2020

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Piezobiocatalysis: Ultrasound-Driven Enzymatic Oxyfunctionalization of C-H Bonds

Yoon, Jaeho (author), Kim, Jinhyun (author), Tieves, F. (author), Zhang, W. (author), Alcalde, Miguel (author), Hollmann, F. (author), Park, Chan Beum (author)

Peroxygenases have long inspired the selective oxyfunctionalization of various aliphatic and aromatic compounds, because of their broad substrate spectrum and simplicity of catalytic mechanism. This study provides a proof-of-concept of piezobiocatalysis by demonstrating peroxygenase-catalyzed oxyfunctionalization reactions fueled by...

journal article 2020

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Benchmarking of laboratory evolved unspecific peroxygenases for the synthesis of human drug metabolites

Gomez de Santos, P. (author), Cervantes, Fadia V. (author), Tieves, F. (author), Plou, Francisco J. (author), Hollmann, F. (author), Alcalde, Miguel (author)

By mimicking the role of human liver P450 monooxygenases, fungal unspecific peroxygenases (UPOs) can perform a range of highly selective oxyfunctionalization reactions on pharmacological compounds, including O-dealkylations and hydroxylations, thereby simulating drug metabolism. Here we have benchmarked human drug metabolite (HDM) synthesis...

journal article 2019

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Light-Harvesting Dye-Alginate Hydrogel for Solar-Driven, Sustainable Biocatalysis of Asymmetric Hydrogenation

Yoon, Jaeho (author), Lee, S.H. (author), Tieves, F. (author), Rauch, M.C.R. (author), Hollmann, F. (author), Park, Chan Beum (author)

We report visible light-driven, asymmetric hydrogenation of C=C bonds using an ene-reductase from Thermus scotoductus SA-01 (TsOYE) and a light-harvesting dye (rose bengal, RB) co-immobilized in an alginate hydrogel. Highly efficient encapsulation of RB in alginate hydrogel was achieved using the intrinsic affinity between TsOYE and RB,...

journal article 2019

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Energising the E-factor: The E⁺-factor

Tieves, F. (author), Tonin, F. (author), Fernandez Fueyo, E. (author), Robbins, John M. (author), Bommarius, Bettina (author), Bommarius, Andreas S. (author), Alcalde, Miguel (author), Hollmann, F. (author)

The E-factor has become an important measure for the environmental impact of (bio)chemical reactions. However, summing up the obvious wastes generated in the laboratory neglects energy-related wastes (mostly greenhouse gases) which are generated elsewhere. To estimate these wastes, we propose to extend the E-factor by an energy-term (E<sup>+<...

journal article 2019

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Expanding the Spectrum of Light-Driven Peroxygenase Reactions

Willot, S.J. (author), Fernandez Fueyo, E. (author), Tieves, F. (author), Pesic, M. (author), Alcalde, Miguel (author), Arends, I.W.C.E. (author), Park, Chan Beum (author), Hollmann, F. (author)

Peroxygenases require a controlled supply of H₂O₂ to operate efficiently. Here, we propose a photocatalytic system for the reductive activation of ambient O₂ to produce H₂O₂ which uses the energy provided by visible light more efficiently based on the combination of wavelength...

journal article 2019

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Bias-Free in Situ H₂O₂ Generation in a Photovoltaic-Photoelectrochemical Tandem Cell for Biocatalytic Oxyfunctionalization

Choi, Da Som (author), Lee, Hojin (author), Tieves, F. (author), Lee, Yang Woo (author), Son, Eun Jin (author), Zhang, W. (author), Shin, Byungha (author), Hollmann, F. (author), Park, Chan Beum (author)

Peroxygenases catalyze selective oxyfunctionalization of hydrocarbons with high conversion efficiencies using H₂O₂ as a key cosubstrate. Here, we report an unbiased photoelectrochemical (PEC) tandem structure consisting of a FeOOH/BiVO₄ photoanode, a Cu(In,Ga)Se₂ solar absorber, and a graphitic...

journal article 2019

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Efficient Aerobic Oxidation of trans-2-Hexen-1-ol using the Aryl Alcohol Oxidase from Pleurotus eryngii

Pedroso de Almeida, T. (author), van Schie, M.M.C.H. (author), Ma, A. (author), Tieves, F. (author), Younes, S.H.H. (author), Fernandez Fueyo, E. (author), Arends, I.W.C.E. (author), Riul, A. (author), Hollmann, F. (author)

The selective oxidation of trans-2-hexen-1-ol to the corresponding aldehyde using a recombinant aryl alcohol oxidase from Pleurotus eryngii (PeAAOx) is reported. Especially using the two liquid phase system to overcome solubility and product inhibition issues enabled to achieve more than 2.200.000 catalytic turnovers for the production enzyme...

journal article 2019

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Chemoenzymatic Halocyclization of γ,δ-Unsaturated Carboxylic Acids and Alcohols

Younes, S.H.H. (author), Tieves, F. (author), Lan, Dongming (author), Wang, Yonghua (author), Süss, Philipp (author), Brundiek, Henrike (author), Wever, Ron (author), Hollmann, F. (author)

A chemoenzymatic method for the halocyclization of unsaturated alcohols and acids by using the robust V-dependent chloroperoxidase from Curvularia inaequalis (CiVCPO) as catalyst has been developed for the in situ generation of hypohalites. A broad range of halolactones and cyclic haloethers are formed with excellent performance of the...

journal article 2019

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Formate Oxidase (FOx) from Aspergillus oryzae: One Catalyst Enables Diverse H ₂ O ₂ -Dependent Biocatalytic Oxidation Reactions

Tieves, F. (author), Willot, S.J. (author), van Schie, M.M.C.H. (author), Rauch, M.C.R. (author), Younes, S.H.H. (author), Zhang, W. (author), Dong, J. (author), Gomez de Santos, P. (author), Hollmann, F. (author)

An increasing number of biocatalytic oxidation reactions rely on H ₂ O ₂ as a clean oxidant. The poor robustness of most enzymes towards H ₂ O ₂ , however, necessitates more efficient systems for in situ H ₂ O ₂ generation. In analogy to the well-known formate dehydrogenase to...

journal article 2019

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Photoenzymatic Hydroxylation of Ethylbenzene Catalyzed by Unspecific Peroxygenase: Origin of Enzyme Inactivation and the Impact of Light Intensity and Temperature

Burek, Bastien O. (author), de Boer, Sabrina R. (author), Tieves, F. (author), Zhang, W. (author), van Schie, M.M.C.H. (author), Bormann, Sebastian (author), Alcalde, Miguel (author), Holtmann, Dirk (author), Hollmann, F. (author)

Photoenzymatic cascades can be used for selective oxygenation of C−H-Bonds under mild conditions circumventing the hydrogen peroxide mediated peroxygenase inactivation via in situ H₂O₂ generation. Here, we report the “on demand” production of hydrogen peroxide via methanol assisted reduction of molecular oxygen using UV...

journal article 2019

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P450BM3-Catalyzed oxidations employing dual functional small molecules

Willot, S.J. (author), Tieves, F. (author), Girhard, Marco (author), Urlacher, Vlada B. (author), Hollmann, F. (author), De Gonzalo, G. (author)

A set of dual functional small molecules (DFSMs) containing different amino acids has been synthesized and employed together with three different variants of the cytochrome P450 monooxygenase P450BM3 from Bacillus megaterium in H2O2-dependent oxidation reactions. These DFSMs enhance P450BM3 activity with hydrogen peroxide as an oxidant,...

journal article 2019

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Nicotinamide adenine dinucleotide as a photocatalyst

Kim, Jinhyun (author), Lee, Sahng Ha (author), Tieves, F. (author), Paul, C.E. (author), Hollmann, F. (author), Park, Chan Beum (author)

Nicotinamide adenine dinucleotide (NAD⁺) is a key redox compound in all living cells responsible for energy transduction, genomic integrity, life-span extension, and neuromodulation. Here, we report a new function of NAD⁺ as a molecular photocatalyst in addition to the biological roles. Our spectroscopic and...

journal article 2019

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Cascading g-C₃N₄ and Peroxygenases for Selective Oxyfunctionalization Reactions

van Schie, M.M.C.H. (author), Zhang, W. (author), Tieves, F. (author), Choi, Da Som (author), Park, Chan Beum (author), Burek, Bastien O. (author), Bloh, Jonathan Z. (author), Arends, I.W.C.E. (author), Paul, C.E. (author), Alcalde, Miguel (author), Hollmann, F. (author)

Peroxygenases are very interesting catalysts for specific oxyfunctionalization chemistry. Instead of relying on complicated electron transport chains, they rely on simple hydrogen peroxide as the stoichiometric oxidant. Their poor robustness against H₂O₂ can be addressed via in situ generation of H₂O<sub>2<...

journal article 2019

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Peroxygenase-Catalysed Epoxidation of Styrene Derivatives in Neat Reaction Media

Rauch, M.C.R. (author), Tieves, F. (author), Paul, C.E. (author), Arends, I.W.C.E. (author), Alcalde, Miguel (author), Hollmann, F. (author)

Biocatalytic oxyfunctionalisation reactions are traditionally conducted in aqueous media limiting their production yield. Here we report the application of a peroxygenase in neat reaction conditions reaching product concentrations of up to 360 mM.

journal article 2019

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Biocatalytic synthesis of the Green Note trans-2-hexenal in a continuous-flow microreactor

van Schie, M.M.C.H. (author), Pedroso de Almeida, T. (author), Laudadio, Gabriele (author), Tieves, F. (author), Fernandez Fueyo, E. (author), Noël, Timothy (author), Arends, I.W.C.E. (author), Hollmann, F. (author)

The biocatalytic preparation of trans-hex-2-enal from trans-hex-2-enol using a novel aryl alcohol oxidase from Pleurotus eryngii (PeAAOx) is reported. As O₂-dependent enzyme PeAAOx-dependent reactions are generally plagued by the poor solubility of O₂ in aqueous media and mass transfer limitations resulting in poor...

journal article 2018

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Biocatalytic C=C Bond Reduction through Carbon Nanodot-Sensitized Regeneration of NADH Analogues

Kim, Jinhyun (author), Lee, S.H. (author), Tieves, F. (author), Choi, Da Som (author), Hollmann, F. (author), Paul, C.E. (author), Park, Chan Beum (author)

Light-driven activation of redox enzymes is an emerging route for sustainable chemical synthesis. Among redox enzymes, the family of Old Yellow Enzyme (OYE) dependent on the nicotinamide adenine dinucleotide cofactor (NADH) catalyzes the stereoselective reduction of α,β-unsaturated hydrocarbons. Here, we report OYE-catalyzed asymmetric...

journal article 2018

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Selective Synthesis of the Human Drug Metabolite 5′-Hydroxypropranolol by an Evolved Self-Sufficient Peroxygenase

Gomez De Santos, Patricia (author), Cañellas, Marina (author), Tieves, F. (author), Younes, S.H.H. (author), Molina-Espeja, Patricia (author), Hofrichter, Martin (author), Hollmann, F. (author), Guallar, Victor (author), Alcalde, Miguel (author)

Propranolol is a widely used beta-blocker that is metabolized by human liver P450 monooxygenases into equipotent hydroxylated human drug metabolites (HDMs). It is paramount for the pharmaceutical industry to evaluate the toxicity and activity of these metabolites, but unfortunately, their synthesis has hitherto involved the use of severe...

journal article 2018