Presentation Title

Cobaltocenium Molecules for Electrode Driven P450 Biocatalysis

Faculty Mentor

Andrew K. Udit

Start Date

17-11-2018 10:15 AM

End Date

17-11-2018 10:30 AM

Location

C323

Session

Oral 2

Type of Presentation

Oral Talk

Subject Area

physical_mathematical_sciences

Abstract

Cytochrome P450 is a class of hemeproteins found in nearly all forms of life. This enzyme is critical to detoxification and bioactivation of many pharmaceuticals. Its use in industry could lead to the ability to manufacture powerful new drug derivatives. Moreover, P450 has the potential to create a safer, more economic method for the testing of new drugs. Current laboratory uses of the enzyme are inefficient and require the use of an expensive mediator molecule called NAD(P)H. This mediator can cost nearly $1,000,000 per mole. NAD(P)H serves as a source of electrons to reduce P450 so that it may function, but cost prohibits its use in many research laboratories. In contrast, cobaltocenium molecules that have the potential to act in place of NAD(P)H can be synthesized cheaply and in large quantities. This research project investigates the synthesis and use of an asymmetric, bifunctional cobaltocenium molecule to serve as an electron relay between an electrode and the enzyme, thus eliminating the need for NAD(P)H. Peptide coupling reagents will be utilized to provide similar functionality to an acid chloride route, while allowing for mild reaction conditions. Preliminary NMR data suggests a mixture of products is obtained following the reaction. Our current work centers on developing a purification method to separate the components of the reaction mixture. Upon isolating the asymmetric derivative, we will begin testing its redox characteristics, as well as its potential as a mediator in P450 biocatalysis.

Summary of research results to be presented

We have synthesized a mixture of homo- and hetero-bifunctionalized cobaltocenium amides. Most cobaltocenium chemistry relies on the precipitation of products from reactants using acid base chemistry or by adding a salt such as NaPF6 to decrease the solubility of the product in the reaction solvent. The product can then be filtered off from the reactants or vice versa. This approach is not applicable for our goal, therefore we have developed a solvent system for the purification of our product over a silica gel column. After separating the products, spectroscopy data (1H-NMR, IR) indicated the presence of the desired functional groups in our products. To further confirm that we had successfully synthesized our product, we performed a copper catalyzed azide-alkyne click reaction with the fluorescent molecule coumarin azide. After workup, the solution containing our cobaltocenium molecule fluoresced, indicating the click reaction was successful and we had in fact synthesized the desired molecule. NMR of this conjugated molecule provided additional evidence suggesting successful synthesis. Overall, our work has resulted in a novel purification method for cobaltocenium molecules, as well as a potential new method for functionalization of cobaltocenium.

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Nov 17th, 10:15 AM Nov 17th, 10:30 AM

Cobaltocenium Molecules for Electrode Driven P450 Biocatalysis

C323

Cytochrome P450 is a class of hemeproteins found in nearly all forms of life. This enzyme is critical to detoxification and bioactivation of many pharmaceuticals. Its use in industry could lead to the ability to manufacture powerful new drug derivatives. Moreover, P450 has the potential to create a safer, more economic method for the testing of new drugs. Current laboratory uses of the enzyme are inefficient and require the use of an expensive mediator molecule called NAD(P)H. This mediator can cost nearly $1,000,000 per mole. NAD(P)H serves as a source of electrons to reduce P450 so that it may function, but cost prohibits its use in many research laboratories. In contrast, cobaltocenium molecules that have the potential to act in place of NAD(P)H can be synthesized cheaply and in large quantities. This research project investigates the synthesis and use of an asymmetric, bifunctional cobaltocenium molecule to serve as an electron relay between an electrode and the enzyme, thus eliminating the need for NAD(P)H. Peptide coupling reagents will be utilized to provide similar functionality to an acid chloride route, while allowing for mild reaction conditions. Preliminary NMR data suggests a mixture of products is obtained following the reaction. Our current work centers on developing a purification method to separate the components of the reaction mixture. Upon isolating the asymmetric derivative, we will begin testing its redox characteristics, as well as its potential as a mediator in P450 biocatalysis.