The use of enzyme-catalyst interfaces is underexplored in the field of biocatalysis, particularly in studies on enabling novel reactivity of enzymes. For this thesis, the HaloTag® protein tagging platform was proposed as a bioconjugation method for a pinacol coupling reaction using lipases, as a model for novel reactivities proceeding via ketyl radical intermediates and hydrogen-bonding-facilitated redox attenuation. After an initial lipase screening of 9 lipases, one lipase (Candida rugosa) was found to perform the pinacol coupling of p-anisaldehyde under standard conditions (fluorescein and 530nm light, 3% yield). Based on a retrosynthetic analysis for the photocatalyst-incorporated HaloTag® linker, the intermediates haloamine 1 and aldehyde 6 were synthesized. Further experiments are underway or planned to complete linker synthesis and conduct pinacol coupling experiments with a bioconjugated system. This project underscores the promising biocatalytic promiscuity of lipases for performing reactions proceeding through ketyl radical intermediates, as well as the underdeveloped potential of incorporating bioengineering principles like bioconjugation into biocatalysis to overcome kinetic barriers to electron transfer and optimize biocatalytic reactions.

Terpene cyclizations are one of the most complex reactions that occur in nature. Scientists have found that replicating this reaction in a lab setting has proved to be immensely challenging as result of the numerous intermediates that must be controlled through the cyclization process in the absence of an enzyme. This study uses commercially available lipases to conduct reactions with geraniol-derived starting materials to identify conditions for performing a terpene cyclization effectively and efficiently. Through hypothesized screening of enzymes and reaction conditions, we have identified a protocol for the successful cyclization of limonene and other geranyl-derived products.