Light-driven decarboxylative deuteration enabled by a divergently engineered photodecarboxylase

Xu, Jian; Fan, Jiajie; Lou, Yujiao; Xu, W.; Wang, Zhiguo; Li, Danyang; Zhou, Haonan; Lin, Xianfu; Wu, Qi

doi:10.1038/s41467-021-24259-6

Cited by 69 publications

(27 citation statements)

References 72 publications

(95 reference statements)

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“…In rapid succession, FRISM has been applied and extended using different enzyme types, each time guides for making rational choices being offered [46] . Prominent examples concern a photodecarboxylase, [46a] and a glycosyltransferase for synthetically difficult regioselective glucosylation of poly‐hydroxy compounds [46b] . Interestingly, around the same time a study appeared in which a glycosyltransferase was also engineered for substrate acceptance (plant flavonoids) using CAST/ISM, the authors noting that the use of FRISM would not have provided their best quadruple mutant [47] .…”

Section: Current Methods Of Rational Enzyme Designmentioning

confidence: 99%

Making Enzymes Suitable for Organic Chemistry by Rational Protein Design

Reetz

2022

ChemBioChem

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This review outlines recent developments in protein engineering of stereo-and regioselective enzymes, which are of prime interest in organic and pharmaceutical chemistry as well as biotechnology. The widespread application of enzymes was hampered for decades due to limited enantio-, diastereo-and regioselectivity, which was the reason why most organic chemists were not interested in biocatalysis. This attitude began to change with the advent of semi-rational directed evolution methods based on focused saturation mutagenesis at sites lining the binding pocket. Screening constitutes the laborintensive step (bottleneck), which is the reason why various research groups are continuing to develop techniques for the generation of small and smart mutant libraries. Rational enzyme design, traditionally an alternative to directed evolution, provides small collections of mutants which require minimal screening. This approach first focused on thermostabilization, and did not enter the field of stereoselectivity until later. Computational guides such as the Rosetta algorithms, HotSpot Wizard metric, and machine learning (ML) contribute significantly to decision making. The newest advancements show that semi-rational directed evolution such as CAST/ISM and rational enzyme design no longer develop on separate tracks, instead, they have started to merge. Indeed, researchers utilizing the two approaches have learned from each other. Today, the toolbox of organic chemists includes enzymes, primarily because the possibility of controlling stereoselectivity by protein engineering has ensured reliability when facing synthetic challenges. This review was also written with the hope that undergraduate and graduate education will include enzymes more so than in the past.[a] M. Reetz Max-Planck-Institut fur Kohlenforschung Mülheim an der Ruhr (Germany)

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Section: Current Methods Of Rational Enzyme Designmentioning

confidence: 99%

Making Enzymes Suitable for Organic Chemistry by Rational Protein Design

Reetz

2022

ChemBioChem

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“…An engineered G462Y mutant of this fatty acid photodecarboxylase from Chlorella variabilis has been shown to provide the unreacted ( R )‐enantiomer in >99 % ee in the kinetic resolution of racemic 2‐hydroxy‐octanoic acid at 10 m m substrate concentration [190] . Decarboxylative deuteration of various aliphatic, aromatic, and bulky cyclic carboxylic acids has been achieved with a high degree of deuterium incorporation using D 2 O and engineered enzyme variants obtained by divergent directed evolution [191] …”

Section: Selective Biocatalytic C‐defunctionalizationmentioning

confidence: 99%

Selective Biocatalytic Defunctionalization of Raw Materials

Wohlgemuth

2022

ChemSusChem

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Biobased raw materials, such as carbohydrates, amino acids, nucleotides, or lipids contain valuable functional groups with oxygen and nitrogen atoms. An abundance of many functional groups of the same type, such as primary or secondary hydroxy groups in carbohydrates, however, limits the synthetic usefulness if similar reactivities cannot be differentiated. Therefore, selective defunctionalization of highly functionalized biobased starting materials to differentially functionalized compounds can provide a sustainable access to chiral synthons, even in case of products with fewer functional groups. Selective defunctionalization reactions, without affecting other functional groups of the same type, are of fundamental interest for biocatalytic reactions. Controlled biocatalytic defunctionalizations of biobased raw materials are attractive for obtaining valuable platform chemicals and building blocks. The biocatalytic removal of functional groups, an important feature of natural metabolic pathways, can also be utilized in a systemic strategy for sustainable metabolite synthesis.

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“…Further improvement of productivity with shorter-chain fatty acids would likely be attainable by using variants of CvFAP. 51 Alternatively, a continuous flow decarboxylation protocol in neat Tris•HCl (100 mM, pH 8.5) buffer was developed, whereby substrate concentration was lowered to 3 mM in order to avoid precipitation. Under optimal conditions, a pentadecane production rate of 0.5 g•L -1 •h -1 (2.5 mmol•L -1 •h -1 ) was achieved.…”

Section: Photodecarboxylation In Flow Using a Homogenous Reaction Mix...mentioning

confidence: 99%

Strategies for Transferring Photobiocatalysis to Continuous Flow Exemplified by the Photodecarboxylation of Fatty Acids

Simić

Jakštaitė

Huck

et al. 2022

Preprint

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The challenges of light-dependent biocatalytic transformations of lipophilic substrates in aqueous media are manifold. For instance, photolability of the catalyst as well as insufficient light penetration into the reaction vessel may be further exacerbated by a heterogeneously dispersed substrate. Light penetration may be addressed by performing the reaction in continuous flow, which allows two modes of applying the catalyst: (i) heterogenously, immobilized on a carrier, which requires light-permeable supports, or (ii) homogenously, dissolved in the reaction mixture. Taking the light-dependent photodecarboxylation of palmitic acid catalyzed by the fatty-acid photodecarboxylase from Chlorella variabilis (CvFAP) as a showcase, strategies for the transfer of a photoenzyme-catalyzed reaction into continuous flow were identified. A range of different supports was evaluated for the immobilization of CvFAP, whereby Eupergit C250 L was the carrier of choice. As the photostability of the catalyst was a limiting factor, a homogeneous system was preferred instead of employing the heterogenized enzyme. This implied that photolabile enzymes may preferably be applied in solution if repair mechanisms cannot be provided. Furthermore, when comparing different wavelengths and light intensities, extinction coefficients may be considered to ensure comparable absorption at each wavelength. Employing homogenous conditions in the CvFAP-catalyzed photodecarboxylation of palmitic acid afforded a space-time yield unsurpassed by any reported batch process (5.7 g/L·h, 26.9 mmol/L·h) for this reaction, demonstrating the advantage of continuous flow in attaining higher productivity of photobiocatalytic processes.

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Light-driven decarboxylative deuteration enabled by a divergently engineered photodecarboxylase

Cited by 69 publications

References 72 publications

Making Enzymes Suitable for Organic Chemistry by Rational Protein Design

Making Enzymes Suitable for Organic Chemistry by Rational Protein Design

Selective Biocatalytic Defunctionalization of Raw Materials

Strategies for Transferring Photobiocatalysis to Continuous Flow Exemplified by the Photodecarboxylation of Fatty Acids

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