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Papers by Daniel Eggerichs

Angewandte Chemie, Mar 16, 2023

Applied and Environmental Microbiology, Feb 12, 2021

Angewandte Chemie, Mar 15, 2023

Angewandte Chemie, Mar 16, 2023

Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktion... more Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktionen. Die dazugehörigen Monooxygenasen der Gruppe E (GEMs) katalysieren enantioselektive Epoxidierungs‐ und Sulfoxidierungsreaktionen. In dieser Arbeit beschreiben wir die Kristallstruktur einer Indol‐Monooxygenase aus dem Bakterium Variovorax paradoxus EPS, einer GEM mit der Bezeichnung VpIndA1. Basierend auf Substratkomplexstrukturen produktiver Bindungsmodi und in Verbindung mit Kraftfeldberechnungen sowie durch schnelle Mischungskinetik zeigen wir die strukturelle Grundlage der Substrat‐ und Stereoselektivität auf. Die strukturbasierte Umgestaltung der Substrattasche führte zu Varianten mit neuer Substratselektivität (für die Sulfoxidation von Benzylphenylsulfid) oder mit stark erhöhter Stereoselektivität (von 35.1 % auf 99.8 % ee für die Herstellung von (1S,2R)‐Indenoxid). Diese erste Bestimmung des Substratbindungsmodus von GEMs in Verbindung mit der Untersuchung von Struktur‐Funktions‐Beziehungen öffnet die Tür für ein strukturbasiertes Design dieser leistungsstarken Biokatalysatoren.

Angewandte Chemie, Mar 15, 2023

Journal of Biological Chemistry

Angewandte Chemie

Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktion... more Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktionen. Die dazugehörigen Monooxygenasen der Gruppe E (GEMs) katalysieren enantioselektive Epoxidierungs‐ und Sulfoxidierungsreaktionen. In dieser Arbeit beschreiben wir die Kristallstruktur einer Indol‐Monooxygenase aus dem Bakterium Variovorax paradoxus EPS, einer GEM mit der Bezeichnung VpIndA1. Basierend auf Substratkomplexstrukturen produktiver Bindungsmodi und in Verbindung mit Kraftfeldberechnungen sowie durch schnelle Mischungskinetik zeigen wir die strukturelle Grundlage der Substrat‐ und Stereoselektivität auf. Die strukturbasierte Umgestaltung der Substrattasche führte zu Varianten mit neuer Substratselektivität (für die Sulfoxidation von Benzylphenylsulfid) oder mit stark erhöhter Stereoselektivität (von 35.1 % auf 99.8 % ee für die Herstellung von (1S,2R)‐Indenoxid). Diese erste Bestimmung des Substratbindungsmodus von GEMs in Verbindung mit der Untersuchung von Struktur‐Funktio...

Applied and Environmental Microbiology, 2021

Predation is an important survival strategy of the widespread myxobacteria, but it remains poorly... more Predation is an important survival strategy of the widespread myxobacteria, but it remains poorly understood on the mechanistic level. Without a basic understanding of how prey cell killing and consumption is achieved, it also remains difficult to investigate the role of predation for the complex myxobacterial lifestyle, reciprocal predator-prey relationships, or the impact of predation on complex bacterial soil communities.

Communications chemistry, Jun 3, 2024

Ene‐reductases allow regio‐ and stereoselective reduction of activated C=C double bonds at the ex... more Ene‐reductases allow regio‐ and stereoselective reduction of activated C=C double bonds at the expense of nicotinamide adenine dinucleotide cofactors [NAD(P)H]. Biological NAD(P)H can be replaced by synthetic mimics to facilitate enzyme screening and process optimization. The ene‐reductase \(\it F\)OYE‐1, originating from an acidophilic iron oxidizer, has been described as a promising candidate and is now being explored for applied biocatalysis. Biological and synthetic nicotinamide cofactors were evaluated to fuel \(\it F\)OYE‐1 to produce valuable compounds. A maximum activity of (319.7\(\pm\)3.2) U mg\(^{−1}\) with NADPH or of (206.7\(\pm\)3.4) U mg\(^{−1}\) with 1‐benzyl‐1,4‐dihydronicotinamide (BNAH) for the reduction of \(\it N\)‐methylmaleimide was observed at 30 °C. Notably, BNAH was found to be a promising reductant but exhibits poor solubility in water. Different organic solvents were therefore assayed: \(\it F\)OYE‐1 showed excellent performance in most systems with up to...

Biotechnology Advances, 2021

Flavoprotein monooxygenases (FPMOs) are single-or two-component enzymes that catalyze a diverse s... more Flavoprotein monooxygenases (FPMOs) are single-or two-component enzymes that catalyze a diverse set of chemo-, regio-and enantioselective oxyfunctionalization reactions. In this review, we describe how FPMOs have evolved from model enzymes in mechanistic flavoprotein research to biotechnologically relevant catalysts that can be applied for the sustainable production of valuable chemicals. After a historical account of the development of the FPMO field, we explain the FPMO classification system, which is primarily based on protein structural properties and electron donor specificities. We then summarize the most appealing reactions catalyzed by each group with a focus on the different types of oxygenation chemistries. Wherever relevant, we report engineering strategies that have been used to improve the robustness and applicability of FPMOs.

Flavin-Dependent Enzymes: Mechanisms, Structures and Applications, 2020

Styrene and indole are naturally occurring compounds, which are also produced and processed by va... more Styrene and indole are naturally occurring compounds, which are also produced and processed by various chemical industries. Thus, it is not surprisingly that microorganisms evolved pathways to detoxify or even to utilize those compounds as carbon sources. Especially, among bacteria several routes are described specifically for the activation and degradation of styrene and indole. Respectively, the initial attack toward these compounds occurs via a flavin-dependent monooxygenase: styrene monooxygenase (SMO) or indole monooxygenase (IMO). In the first place, SMOs have been described to initiate a styrene specific degradation. These are in general two-component systems, whereas a small FAD-reductase (SMOB) delivers reduced FAD on the expense of NADH toward the monooxygenase (SMOA). Various modes of interaction are possible and for both mostly dimeric protein subunits structural data were reported. Thus, this flavoprotein monooxygenase-especially the one from Pseudomonas putida S12 can be seen as the prototype of this class of enzymes. In the course of describing related members of this enzyme family some remarkable findings were made. For example, self-sufficient fusion proteins have been reported as well as enzymes, which could not be assigned to a styrene metabolic activity, rather to indole conversion. Later it was found that this flavoprotein group can be separated at least into two subgroups: styrene and indole monooxygenases. And both enzymes rely on a FAD-reductase to obtain the reduced cofactor (FADred), which is employed to activate molecular oxygen toward hydroperoxy-FAD, which allows substrate epoxidation and the formation of hydroxy-FAD, which finally yields H2O and oxidized FAD.

ChemBioChem, 2019

This articlei sp art of aj oint Special Collection dedicated to the Biotrans 2019 symposium. To v... more This articlei sp art of aj oint Special Collection dedicated to the Biotrans 2019 symposium. To view the completecollection, visit our homepage Scheme1.Stereoselective reduction of (R)-carvonethrought he action of an ene-reductase(ER). The nicotinamide (biological or synthetic, NA) actsasa n electrondonor to reduce the flavin cofactor FMN of the ER;this subsequently allows the transfer of ah ydride to C a of the unsaturated substrate. Aproton from ac onserved Tyrr esidue in the ER is added to C b to yield (2R,5R)-dihydrocarvone.

Catalysts, 2020

Styrene and indole monooxygenases (SMO and IMO) are two-component flavoprotein monooxygenases com... more Styrene and indole monooxygenases (SMO and IMO) are two-component flavoprotein monooxygenases composed of a nicotinamide adenine dinucleotide (NADH)-dependent flavin adenine dinucleotide (FAD)-reductase (StyB or IndB) and a monooxygenase (StyA or IndA). The latter uses reduced FAD to activate oxygen and to oxygenate the substrate while releasing water. We circumvented the need for the reductase by direct FAD reduction in solution using the NAD(P)H-mimic 1-benzyl-1,4-dihydronicotinamide (BNAH) to fuel monooxygenases without NADH requirement. Herein, we report on the hitherto unknown solvent tolerance for the indole monooxygenase from Gemmobacter nectariphilus DSM15620 (GnIndA) and the styrene monooxygenase from Gordonia rubripertincta CWB2 (GrStyA). These enzymes were shown to convert bulky and rather hydrophobic styrene derivatives in the presence of organic cosolvents. Subsequently, BNAH-driven biotransformation was furthermore optimized with regard to the applied cosolvent and its...

Angewandte Chemie, Mar 16, 2023

Applied and Environmental Microbiology, Feb 12, 2021

Angewandte Chemie, Mar 15, 2023

Angewandte Chemie, Mar 16, 2023

Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktion... more Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktionen. Die dazugehörigen Monooxygenasen der Gruppe E (GEMs) katalysieren enantioselektive Epoxidierungs‐ und Sulfoxidierungsreaktionen. In dieser Arbeit beschreiben wir die Kristallstruktur einer Indol‐Monooxygenase aus dem Bakterium Variovorax paradoxus EPS, einer GEM mit der Bezeichnung VpIndA1. Basierend auf Substratkomplexstrukturen produktiver Bindungsmodi und in Verbindung mit Kraftfeldberechnungen sowie durch schnelle Mischungskinetik zeigen wir die strukturelle Grundlage der Substrat‐ und Stereoselektivität auf. Die strukturbasierte Umgestaltung der Substrattasche führte zu Varianten mit neuer Substratselektivität (für die Sulfoxidation von Benzylphenylsulfid) oder mit stark erhöhter Stereoselektivität (von 35.1 % auf 99.8 % ee für die Herstellung von (1S,2R)‐Indenoxid). Diese erste Bestimmung des Substratbindungsmodus von GEMs in Verbindung mit der Untersuchung von Struktur‐Funktions‐Beziehungen öffnet die Tür für ein strukturbasiertes Design dieser leistungsstarken Biokatalysatoren.

Angewandte Chemie, Mar 15, 2023

Journal of Biological Chemistry

Angewandte Chemie

Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktion... more Flavoprotein‐Monooxygenasen sind eine vielseitige Gruppe von Enzymen für biokatalytische Reaktionen. Die dazugehörigen Monooxygenasen der Gruppe E (GEMs) katalysieren enantioselektive Epoxidierungs‐ und Sulfoxidierungsreaktionen. In dieser Arbeit beschreiben wir die Kristallstruktur einer Indol‐Monooxygenase aus dem Bakterium Variovorax paradoxus EPS, einer GEM mit der Bezeichnung VpIndA1. Basierend auf Substratkomplexstrukturen produktiver Bindungsmodi und in Verbindung mit Kraftfeldberechnungen sowie durch schnelle Mischungskinetik zeigen wir die strukturelle Grundlage der Substrat‐ und Stereoselektivität auf. Die strukturbasierte Umgestaltung der Substrattasche führte zu Varianten mit neuer Substratselektivität (für die Sulfoxidation von Benzylphenylsulfid) oder mit stark erhöhter Stereoselektivität (von 35.1 % auf 99.8 % ee für die Herstellung von (1S,2R)‐Indenoxid). Diese erste Bestimmung des Substratbindungsmodus von GEMs in Verbindung mit der Untersuchung von Struktur‐Funktio...

Applied and Environmental Microbiology, 2021

Predation is an important survival strategy of the widespread myxobacteria, but it remains poorly... more Predation is an important survival strategy of the widespread myxobacteria, but it remains poorly understood on the mechanistic level. Without a basic understanding of how prey cell killing and consumption is achieved, it also remains difficult to investigate the role of predation for the complex myxobacterial lifestyle, reciprocal predator-prey relationships, or the impact of predation on complex bacterial soil communities.

Communications chemistry, Jun 3, 2024

Ene‐reductases allow regio‐ and stereoselective reduction of activated C=C double bonds at the ex... more Ene‐reductases allow regio‐ and stereoselective reduction of activated C=C double bonds at the expense of nicotinamide adenine dinucleotide cofactors [NAD(P)H]. Biological NAD(P)H can be replaced by synthetic mimics to facilitate enzyme screening and process optimization. The ene‐reductase \(\it F\)OYE‐1, originating from an acidophilic iron oxidizer, has been described as a promising candidate and is now being explored for applied biocatalysis. Biological and synthetic nicotinamide cofactors were evaluated to fuel \(\it F\)OYE‐1 to produce valuable compounds. A maximum activity of (319.7\(\pm\)3.2) U mg\(^{−1}\) with NADPH or of (206.7\(\pm\)3.4) U mg\(^{−1}\) with 1‐benzyl‐1,4‐dihydronicotinamide (BNAH) for the reduction of \(\it N\)‐methylmaleimide was observed at 30 °C. Notably, BNAH was found to be a promising reductant but exhibits poor solubility in water. Different organic solvents were therefore assayed: \(\it F\)OYE‐1 showed excellent performance in most systems with up to...

Biotechnology Advances, 2021

Flavoprotein monooxygenases (FPMOs) are single-or two-component enzymes that catalyze a diverse s... more Flavoprotein monooxygenases (FPMOs) are single-or two-component enzymes that catalyze a diverse set of chemo-, regio-and enantioselective oxyfunctionalization reactions. In this review, we describe how FPMOs have evolved from model enzymes in mechanistic flavoprotein research to biotechnologically relevant catalysts that can be applied for the sustainable production of valuable chemicals. After a historical account of the development of the FPMO field, we explain the FPMO classification system, which is primarily based on protein structural properties and electron donor specificities. We then summarize the most appealing reactions catalyzed by each group with a focus on the different types of oxygenation chemistries. Wherever relevant, we report engineering strategies that have been used to improve the robustness and applicability of FPMOs.

Flavin-Dependent Enzymes: Mechanisms, Structures and Applications, 2020

Styrene and indole are naturally occurring compounds, which are also produced and processed by va... more Styrene and indole are naturally occurring compounds, which are also produced and processed by various chemical industries. Thus, it is not surprisingly that microorganisms evolved pathways to detoxify or even to utilize those compounds as carbon sources. Especially, among bacteria several routes are described specifically for the activation and degradation of styrene and indole. Respectively, the initial attack toward these compounds occurs via a flavin-dependent monooxygenase: styrene monooxygenase (SMO) or indole monooxygenase (IMO). In the first place, SMOs have been described to initiate a styrene specific degradation. These are in general two-component systems, whereas a small FAD-reductase (SMOB) delivers reduced FAD on the expense of NADH toward the monooxygenase (SMOA). Various modes of interaction are possible and for both mostly dimeric protein subunits structural data were reported. Thus, this flavoprotein monooxygenase-especially the one from Pseudomonas putida S12 can be seen as the prototype of this class of enzymes. In the course of describing related members of this enzyme family some remarkable findings were made. For example, self-sufficient fusion proteins have been reported as well as enzymes, which could not be assigned to a styrene metabolic activity, rather to indole conversion. Later it was found that this flavoprotein group can be separated at least into two subgroups: styrene and indole monooxygenases. And both enzymes rely on a FAD-reductase to obtain the reduced cofactor (FADred), which is employed to activate molecular oxygen toward hydroperoxy-FAD, which allows substrate epoxidation and the formation of hydroxy-FAD, which finally yields H2O and oxidized FAD.

ChemBioChem, 2019

This articlei sp art of aj oint Special Collection dedicated to the Biotrans 2019 symposium. To v... more This articlei sp art of aj oint Special Collection dedicated to the Biotrans 2019 symposium. To view the completecollection, visit our homepage Scheme1.Stereoselective reduction of (R)-carvonethrought he action of an ene-reductase(ER). The nicotinamide (biological or synthetic, NA) actsasa n electrondonor to reduce the flavin cofactor FMN of the ER;this subsequently allows the transfer of ah ydride to C a of the unsaturated substrate. Aproton from ac onserved Tyrr esidue in the ER is added to C b to yield (2R,5R)-dihydrocarvone.

Catalysts, 2020

Styrene and indole monooxygenases (SMO and IMO) are two-component flavoprotein monooxygenases com... more Styrene and indole monooxygenases (SMO and IMO) are two-component flavoprotein monooxygenases composed of a nicotinamide adenine dinucleotide (NADH)-dependent flavin adenine dinucleotide (FAD)-reductase (StyB or IndB) and a monooxygenase (StyA or IndA). The latter uses reduced FAD to activate oxygen and to oxygenate the substrate while releasing water. We circumvented the need for the reductase by direct FAD reduction in solution using the NAD(P)H-mimic 1-benzyl-1,4-dihydronicotinamide (BNAH) to fuel monooxygenases without NADH requirement. Herein, we report on the hitherto unknown solvent tolerance for the indole monooxygenase from Gemmobacter nectariphilus DSM15620 (GnIndA) and the styrene monooxygenase from Gordonia rubripertincta CWB2 (GrStyA). These enzymes were shown to convert bulky and rather hydrophobic styrene derivatives in the presence of organic cosolvents. Subsequently, BNAH-driven biotransformation was furthermore optimized with regard to the applied cosolvent and its...