Thomas S. Moody

Engineering and application of P450 monooxygenases in pharmaceutical and metabolite synthesis

Cytochrome P450 monoxygenase (P450s or CYPs) allow access to drug metabolites, necessary for approval of new therapeutics in one step, with increased success being demonstrated using bacterial and fungal P450s. Moreover, 12 of the 13 products of the human metabolism of verapamil can be accessed through engineered and chimeric bacterial P450s. These P450s are also used in the synthesis of pharmaceuticals themselves, including the semi-synthetic production of artemisinin in an engineered cell. The integration of new technologies including ultrasound and polyfluorinated hydrocarbon solvents offers an attractive means by the true synthetic potential of ubiquitous P450s can be fully realised.

A metagenomics approach for new biocatalyst discovery: application to transaminases and the synthesis of allylic amines

Transaminase enzymes have significant potential for the sustainable synthesis of amines using mild aqueous reaction conditions. Here a metagenomics mining strategy has been used for new transaminase enzyme discovery. Starting from oral cavity microbiome samples, DNA sequencing and bioinformatics analyses were performed. Subsequent in silico mining of a library of contiguous reads built from the sequencing data identified 11 putative Class III transaminases which were cloned and overexpressed. Several screening protocols were used and three enzymes selected of interest due to activities towards substrates covering a wide structural diversity. Transamination of functionalized cinnamaldehydes was then investigated for the production of valuable amine building blocks.

CHAPTER 8:Almac: An Industrial Perspective of Ene Reductase (ERED) Biocatalysis

The asymmetric hydrogenation of α,β-unsaturated compounds using ene-reductase enzymes (primarily from the ‘Old Yellow Enzyme’ family of flavoproteins) is moving from an academic curiosity to a technology with potential industrial relevance. Continuing identification and reporting of ene-reductases with improved stability properties and higher turnover numbers will ensure their incorporation into industrial processes. This chapter highlights literature examples of bioreductions available to the modern practical chemist.