Peroxygenase‐Catalysed Epoxidation of Styrene Derivatives in Neat Reaction Media

Rauch, Marine; Tieves, Florian; Paul, Caroline E.; Arends, Isabel W. C. E.; Alcalde, Miguel; Hollmann, Frank

doi:10.1002/cctc.201901142

Cited by 47 publications

(31 citation statements)

References 40 publications

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“…Previously reported data for Aae UPO revealed up to 10000 TONs with 4.6% ee when using a light-driven in situ hydrogen peroxide formation 65 . Combining Aae UPO* and tert-butylhydroperoxide added via a syringe pump setup resulted in 3200 TONs and 12% ee 66 . Within our setup, when using Cgl UPO similar epoxidation activities with TONs of 4120 were obtained.…”

Section: Discussionmentioning

confidence: 99%

A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases

et al. 2021

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Fungal unspecific peroxygenases (UPOs) represent an enzyme class catalysing versatile oxyfunctionalisation reactions on a broad substrate scope. They are occurring as secreted, glycosylated proteins bearing a haem-thiolate active site and rely on hydrogen peroxide as the oxygen source. However, their heterologous production in a fast-growing organism suitable for high throughput screening has only succeeded once—enabled by an intensive directed evolution campaign. We developed and applied a modular Golden Gate-based secretion system, allowing the first production of four active UPOs in yeast, their one-step purification and application in an enantioselective conversion on a preparative scale. The Golden Gate setup was designed to be universally applicable and consists of the three module types: i) signal peptides for secretion, ii) UPO genes, and iii) protein tags for purification and split-GFP detection. The modular episomal system is suitable for use in Saccharomyces cerevisiae and was transferred to episomal and chromosomally integrated expression cassettes in Pichia pastoris. Shake flask productions in Pichia pastoris yielded up to 24 mg/L secreted UPO enzyme, which was employed for the preparative scale conversion of a phenethylamine derivative reaching 98.6 % ee. Our results demonstrate a rapid, modular yeast secretion workflow of UPOs yielding preparative scale enantioselective biotransformations.

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Section: Discussionmentioning

confidence: 99%

A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases

et al. 2021

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“… 55 − 58 From a synthetic point of view, r Aae UPO is very interesting as it catalyzes the specific oxyfunctionalization of a broad range of compounds. 7 , 10 , 55 − 57 , 59 − 69 As a heme-dependent enzyme, it however, is also prone to irreversible oxidative inactivation by H 2 O 2 or other reactive oxygen species. Therefore, we next performed an experiment on the hydroxylation of ethylbenzene using r Aae UPO and SAS-mediated H 2 O 2 generation ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

Water-Soluble Anthraquinone Photocatalysts Enable Methanol-Driven Enzymatic Halogenation and Hydroxylation Reactions

et al. 2020

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Peroxyzymes simply use H 2 O 2 as a cosubstrate to oxidize a broad range of inert C–H bonds. The lability of many peroxyzymes against H 2 O 2 can be addressed by a controlled supply of H 2 O 2 , ideally in situ. Here, we report a simple, robust, and water-soluble anthraquinone sulfonate (SAS) as a promising organophotocatalyst to drive both haloperoxidase-catalyzed halogenation and peroxygenase-catalyzed oxyfunctionalization reactions. Simple alcohols, methanol in particular, can be used both as a cosolvent and an electron donor for H 2 O 2 generation. Very promising turnover numbers for the biocatalysts of up to 318 000 have been achieved.

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“…Besides bioreduction processes catalyzed by ADHs, other reactions employing neat conditions have been developed employing oxidative enzymes. Thus, recent examples with the newly discovered fungal unspecific peroxygenases (UPOs) have also been reported by Hollmann's group [46,47]. UPOs are promising enzymes able to perform a wide set of oxyfunctionalization reactions acting as 'self-sufficient' monooxygenases only requiring H 2 O 2 for activation [48,49].…”

Section: Biocatalyzed Redox Processes At Neat Conditionsmentioning

confidence: 98%

“…The same enzyme was later proved to selectively epoxidize a set of styrene derivatives in the neat substrates [ 47 ]. Thus, it was applied to build an interesting chemoenzymatic process to afford the valuable drug (pseudo)ephedrine 9 ( Scheme 4 ).…”

Section: Biocatalyzed Redox Processes At Neat Conditionsmentioning

confidence: 99%

Biocatalyzed Redox Processes Employing Green Reaction Media

Aranda

Gonzalo

2020

Molecules

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The application of biocatalysts to perform reductive/oxidative chemical processes has attracted great interest in recent years, due to their environmentally friendly conditions combined with high selectivities. In some circumstances, the aqueous buffer medium normally employed in biocatalytic procedures is not the best option to develop these processes, due to solubility and/or inhibition issues, requiring biocatalyzed redox procedures to circumvent these drawbacks, by developing novel green non-conventional media, including the use of biobased solvents, reactions conducted in neat conditions and the application of neoteric solvents such as deep eutectic solvents.

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Peroxygenase‐Catalysed Epoxidation of Styrene Derivatives in Neat Reaction Media

Abstract: 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.

Cited by 47 publications

References 40 publications

A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases

A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases

Water-Soluble Anthraquinone Photocatalysts Enable Methanol-Driven Enzymatic Halogenation and Hydroxylation Reactions

Biocatalyzed Redox Processes Employing Green Reaction Media

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