RewiringSaccharomyces cerevisiaemetabolism for optimised Taxol® precursors production

Abstract

Saccharomyces cerevisiae has been recognised as a convenient host for the production of early precursors to the Taxol anticancer drug. Recent studies have highlighted the harmful impact of oxidative stress as a result of the activity of Taxol first cytochrome P450-reductase enzymes (Taxus spp. CYP725A4-POR). Here, we evolved a new oxidative stress-tolerant yeast strain on galactose, which led to a three-fold higher titre of the CYP725A4 enzyme substrate, taxadiene. We comprehensively analysed the performance of the evolved and parent strain in galactose-limited chemostat cultures before and during oxidative stress induction. Integrating the transcriptomics and metabolite profiling data in an enzyme-constrained genome scale model enabled a more accurate prediction of changes that occurred to biological pathways as a response to/consequence of evolution and oxidative stress. The analyses showed a better performance of the evolved strain with improved respiration and reduced overflow metabolites production. The strain was robust to re-introduction of the oxidative stress, potentially due to the cross-protection mechanism, which contributed to likely better heme, flavin and NADPH availability for an optimal expression of CYP725A4 and POR in yeast. The increased level of taxadiene production has potentially occurred due to the antioxidant properties of taxadiene or as a mechanism to overcome the toxicity of geranylgeranyl diphosphate, the precursor to taxadiene synthase.

The synergetic effect from the combination of different adsorption resins in batch and semi‐continuous cultivations of S. Cerevisiae cell factories to produce acetylated Taxanes precursors of the anticancer drug Taxol

“…Microbial cell factories are becoming an important route for the sustainable production of different natural products and chemicals of different commercial (therapeutics, food additives, cosmetics, or biofuels) or academic relevance (Malcı et al, 2020;Walls, Otoupal, et al, 2022;Wong et al, , 2018. To date, a majority of studies involving an integrated process of product synthesis and recovery have not been performed at a lab scale, which has contributed to the slow translation of microbial cell factories to fulfill industry demands (Malcı et al, 2023;Nowrouzi et al, 2023;Walls, Martinez & del Rio Chanona, & Rios-Solis, 2022;. Different bioprocess strategies have been used to enhance the recovery of desired products, for instance, using two or more phase partitions, foam phase recovery or in situ product recovery (ISPR) (Zainuddin et al, 2021).…”

Improved production of Taxol® precursors in S. cerevisiae using combinatorial in silico design and metabolic engineering

“…S. cerevisiae has been harnessed as a platform to produce high-value biopharmaceuticals, ranging from recombinant therapeutic proteins to plant-derived natural products [ 10 – 14 ]. By integrating heterologous plant-derived genes, early-step precursors of Taxol ® (paclitaxel), a leading anticancer drug with a market size over billions of USD [ 15 ], have been produced by yeast cell factories [ 16 – 20 ]. The biosynthesis of Taxol ® initiates with the conversion of geranylgeranyl diphosphate (GGPP), a product of the yeast mevalonate pathway, into taxadiene (taxa-4(5),11(12)-diene) catalyzed by taxadiene synthase [ 21 ].…”

Improved Production of Taxol® Precursors inS. cerevisiaeusing Combinatorialin silicoDesign and Metabolic Engineering

“…cerevisiae has been used as a platform to produce high-value biopharmaceuticals, ranging from recombinant therapeutic proteins to plant-derived natural products [10,11]. By integrating heterologous plant-derived genes, early-step precursors of Taxol® (paclitaxel), a leading anticancer drug with a market size over billions of USD [12], have been produced by yeast cell factories [13][14][15][16][17]. Geranylgeranyl diphosphate (GGPP), a product of the yeast mevalonate pathway, is the substrate of the cyclisation step, the first step in the Taxol® biosynthesis pathway catalyzed by the taxadiene synthase [18].…”