Alkyl Formate Ester Synthesis by a Fungal Baeyer–Villiger Monooxygenase

Ferroni, Felix Martín; Tolmie, Carmien; Smit, Martha S.; Opperman, Diederik J.

doi:10.1002/cbic.201600684

Cited by 17 publications

(12 citation statements)

References 25 publications

(83 reference statements)

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“…However, the formation of the abnormal product by BVMOs was reported for a number of reactions, such as the conversions of chiral nitrilolactones, terpenones, β‐amino ketones, β‐hydroxy ketones, and bicyclic ketones . Divergent regioselectivities for different BVMOs have also been observed for the conversion of aldehydes into formates or carboxylic acids and long‐chain ketocarboxylic acids . Thus, understanding how regioselectivity can be controlled is necessary for the development of tailored BVMOs for a variety of applications.…”

Section: Introductionmentioning

confidence: 99%

Controlling the Regioselectivity of Baeyer–Villiger Monooxygenases by Mutation of Active‐Site Residues

2017

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Baeyer-Villiger monooxygenase (BVMO)-mediated regiodivergent conversions of asymmetric ketones can lead to the formation of "normal" or "abnormal" lactones. In a previous study, we were able to change the regioselectivity of a BVMO by mutation of the active-site residues to smaller amino acids, which thus created more space. In this study, we demonstrate that this method can also be used for other BVMO/substrate combinations. We investigated the regioselectivity of 2-oxo-Δ -4,5,5-trimethylcyclopentenylacetyl-CoA monooxygenase from Pseudomonas putida (OTEMO) for cis-bicyclo[3.2.0]hept-2-en-6-one (1) and trans-dihydrocarvone (2), and we were able to switch the regioselectivity of this enzyme for one of the substrate enantiomers. The OTEMO wild-type enzyme converted (-)-1 into an equal (50:50) mixture of the normal and abnormal products. The F255A/F443V variant produced 90 % of the normal product, whereas the W501V variant formed up to 98 % of the abnormal product. OTEMO F255A exclusively produced the normal lactone from (+)-2, whereas the wild-type enzyme was selective for the production of the abnormal product. The positions of these amino acids were equivalent to those mutated in the cyclohexanone monooxygenases from Arthrobacter sp. and Acinetobacter sp. (CHMO and CHMO ) to switch their regioselectivity towards (+)-2, which suggests that there are hot spots in the active site of BVMOs that can be targeted with the aim to change the regioselectivity.

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Section: Introductionmentioning

confidence: 99%

Controlling the Regioselectivity of Baeyer–Villiger Monooxygenases by Mutation of Active‐Site Residues

2017

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“…In the interest of our study, we performed a screening with group B flavoprotein monooxygenases Type I BVMOs for the selective oxidation of two substrates with BNAH. However, we observed no conversion with the purified wild type CHMO, mutant CHMO M16 , [24] and BVMO 791 [25] (SI section 2.7).…”

Section: Activity Of Type I Bvmos With Ncbsmentioning

confidence: 90%

“…In this respect, the wild‐type cyclohexanone monooxygenase (CHMO) from Acinetobacter sp . NCIMB9871, one mutant (CHMO M16 ), and one BVMO from Aspergillus flavus (BVMO 791 ) were selected.…”

Section: Introductionmentioning

confidence: 99%

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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“…Bayer-Villiger monooxygenase (BVMO) is a avin-dependent enzyme that catalyzes the regioselective Bayer-Villiger oxidation of ketones to the corresponding esters or lactones [Fürst et al 2019;Romero et al 2018;de Gonzalo et al 2010]. BVMOs show broad substrate acceptance of aliphatic ketones [Forney et al 1969;Yu et al 2018;Fiorentini et al 2017] and aromatic extended ketones [van Beek et al 2012;Fraaije et al 2005], and also catalyze other oxidation reactions, including hydroxylation [Ferroni et al 2017;Bisagni et al 2014], epoxidation [Colonna et al 2002;Rial et al 2008], and sulfoxidation [Branchaud et al 1985;de Gonzalo et al 2017]. As BVMO catalysis reactions have high enantio-, regio-, and chemo-selectivity, involve a "green" oxidant (oxygen), and utilize the easily recycled NAD(P)H [Mordhorst et al 2020] as the electronic donor, BVMOs are attracting increasing attention for their potential utility in environmentally benign bio-oxidation processes.…”

Section: Introductionmentioning

confidence: 99%

Secretory Expression of Cyclohexanone Monooxygenase by Methylotrophic Yeast for Efficient Omeprazole Sulfide Bio-oxidation

Zheng

Geng

Liu

et al. 2021

Preprint

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Prochiral pyrmetazole can be asymmetrically oxidized into (S)-omeprazole, a proton pump inhibitor that is used to treat gastroesophageal reflux, by an engineered cyclohexanone monooxygenase (CHMOAcineto-Mut) that has high stereoselectivity. CHMOAcineto-Mut is produced by heterologous expression in Escherichia coli, where it is expressed intracellularly. Thus, isolating this useful biocatalyst requires tedious cell disruption and subsequent purification, which hinders its use for industrial purposes. Here, we report the extracellular production of CHMOAcineto-Mut by a methylotrophic yeast, Pichia pastoris, for the first time. The recombinant CHMOAcineto-Mut expressed by P. pastoris showed a higher flavin occupation rate than that produced by E. coli, and this was accompanied by a 3.2-fold increase in catalytic efficiency. At a cell density of 150 g/L cell dry weight, we achieved a recombinant CHMOAcineto-Mut production rate of 1,700 U/L, representing approximately 85% of the total protein secreted into the fermentation broth. By directly employing the pH adjusted supernatant as a biocatalyst, we were able to almost completely transform 10 g/L of pyrmetazole into the corresponding (S)-sulfoxide, with >99% enantiomeric excess.

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Alkyl Formate Ester Synthesis by a Fungal Baeyer–Villiger Monooxygenase

Cited by 17 publications

References 25 publications

Controlling the Regioselectivity of Baeyer–Villiger Monooxygenases by Mutation of Active‐Site Residues

Controlling the Regioselectivity of Baeyer–Villiger Monooxygenases by Mutation of Active‐Site Residues

Flavoenzyme‐mediated Regioselective Aromatic Hydroxylation with Coenzyme Biomimetics

Secretory Expression of Cyclohexanone Monooxygenase by Methylotrophic Yeast for Efficient Omeprazole Sulfide Bio-oxidation

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