Cytochrome P450-mediated N-demethylation of noscapine by whole-cell biotransformation: process limitations and strategies for optimisation
Luke Richards, Ailsa Jarrold, Tim Bowser, Geoffrey W Stevens, Sally L Gras
Journal of Industrial Microbiology and Biotechnology | Springer Verlag | Published : 2020
Cytochrome P450 enzymes catalyse reactions of significant industrial interest but are underutilised in large-scale bioprocesses due to enzyme stability, cofactor requirements and the poor aqueous solubility and microbial toxicity of typical substrates and products. In this work, we investigate the potential for preparative-scale N-demethylation of the opium poppy alkaloid noscapine by a P450BM3 (CYP102A1) mutant enzyme in a whole-cell biotransformation system. We identify and address several common limitations of whole-cell P450 biotransformations using this model N-demethylation process. Mass transfer into Escherichia coli cells was found to be a major limitation of biotransformation rate a..View full abstract
The authors would like to thank the Mass Spectrometry and Proteomics Facility at the Bio21 Institute (Melbourne) for technical assistance and access to analytical equipment. This work was supported by Sun Pharmaceutical Industries Ltd and The University of Melbourne. LR was supported by an Australian Government Research Training Program Scholarship.