Optimization of the Functional Expression of Plant Cytochrome P450s in Yeast

Development of microbial cell factories holds the potential to enable sustainable biobased production of a wide variety of valuable chemical compounds. Engineering microbial hosts for the biosynthesis of plant natural products often includes expression of cytochrome P450s. Heterologous expression of plant P450s in microbial hosts is known to be challenging, including the requirement for coexpression of redox partners. In order to support biobased production of plant natural products in yeast cell factories we have designed, built and tested a multi-parametric set-up to uncover principles related to optimal enzymatic conversion of a P450-mediated reaction. The parameters of interest include copy number, expression tuning, choice of redox partner and cytochrome B5. Here we present the results of our combinatorial approach using a candidate reaction of relevance to development of platform cell factories for plant natural products.