Selektive Biokatalytische Synthese von Glycerolethern
Biochemie
Zusammenfassung der Projektergebnisse
Recent years have seen an unprecedented expansion of the biocatalytic toolbox, enabling chemical transformations with increased selectivity and under milder conditions compared to their classic chemical counterparts. However, there exists a notable lack of biocatalytic approaches for several fundamental organo-chemical reactions and functional groups. This gap is particularly striking for ethers, which present a prevalent structural motif across pharmaceuticals, agrochemicals, natural products and cosmetics. Although ethers are common in secondary metabolism, they are an underrepresented functional group in primary metabolism. Indeed, nature seemingly avoids ethers for routine applications and, hence, there are comparably few enzymes capable of constructing ether bonds in a general fashion. However, such enzymes are highly sought after for synthetic applications as they typically operate with higher regioselectivity and under milder conditions than traditional organo-chemical approaches. To expand the repertoire of well characterized ether synthases, we explored the catalytic potential of the scarcely researched family of geranylgeranylglyceryl phosphate synthases (G3PSs). In this project, we developed methods for the kinetic examination of these enzymes in high throughput as well as NMR spectroscopic methods to investigate their selectivity. We then applied these methods to study a representative model G3PSs. During our work, we found that this ultrastable enzyme performs otherwise challenging transformations with perfect selectivity. To our surprise, it also proved capable of converting several different substrates, for instance ones featuring much a shorter or highly flexible chain than the native substrate or the opposite alkene configuration. Through mechanistic studies, we learned that G3PSs break most of the paradigms inherent to prenyltransferases, making this enzyme family an intriguing mechanistic outlier. Combined, this project provides the fundamental understanding and methods to introduce G3PSs into the biocatalytic alkylation toolbox. To realize the full potential of these enzymes, future work will explore how their performance can be fine-tuned by protein engineering and develop biocatalytic cascades for the efficient delivery of their substrates from inexpensive starting materials.
Projektbezogene Publikationen (Auswahl)
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Alternative Assay Reagents for UV-Spectroscopic Detection of (Pyro-)Phosphate with the PUB Module. Analytical Chemistry, 94(23), 8132-8135.
Kaspar, Felix; Ganskow, Charity S. G.; Eilert, Lea; Klahn, Philipp & Schallmey, Anett
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UV-Spectroscopic Detection of (Pyro-)Phosphate with the PUB Module. Analytical Chemistry, 94(8), 3432-3435.
Eilert, Lea; Schallmey, Anett & Kaspar, Felix
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Biased Borate Esterification during Nucleoside Phosphorylase‐Catalyzed Reactions: Apparent Equilibrium Shifts and Kinetic Implications**. Angewandte Chemie International Edition, 62(20).
Kaspar, Felix; Brandt, Felix; Westarp, Sarah; Eilert, Lea; Kemper, Sebastian; Kurreck, Anke; Neubauer, Peter; Jacob, Christoph R. & Schallmey, Anett
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Biocatalytic Ether Lipid Synthesis by an Archaeal Glycerolprenylase. Angewandte Chemie International Edition, 63(46).
Kaspar, Felix; Eilert, Lea; Staar, Sophie; Oung, Sangwar Wadtey; Wolter, Mario; Ganskow, Charity S. G.; Kemper, Sebastian; Klahn, Philipp; Jacob, Christoph R.; Blankenfeldt, Wulf & Schallmey, Anett
