Enhanced production of taxadiene in Saccharomyces cerevisiae

Behnaz Nowrouzi, Rachel A. Li, Laura E. Walls, Leo d’Espaux, Koray Malcı, Lungang Liang, Nestor Jonguitud-Borrego, Albert I. Lerma-Escalera, Jose R. Morones-Ramirez, Jay D. Keasling, Leonardo Rios-Solis*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Background: Cost-effective production of the highly effective anti-cancer drug, paclitaxel (Taxol®), remains limited despite growing global demands. Low yields of the critical taxadiene precursor remains a key bottleneck in microbial production. In this study, the key challenge of poor taxadiene synthase (TASY) solubility in S. cerevisiae was revealed, and the strains were strategically engineered to relieve this bottleneck.
Results: Multi-copy chromosomal integration of TASY harbouring a selection of fusion solubility tags improved taxadiene titres 22-fold, up to 57 ± 3 mg/L at 30 °C at microscale, compared to expressing a single episomal copy of TASY. The scalability of the process was highlighted through achieving similar titres during scale up to 25 mL and 250 mL in shake flask and bioreactor cultivations, respectively at 20 and 30 °C. Maximum taxadiene titres of 129 ± 15 mg/L and 127 mg/L were achieved through shake flask and bioreactor cultivations, respectively, of the optimal strain at a reduced temperature of 20 °C.
Conclusions: The results of this study highlight the benefit of employing a combination of molecular biology and bioprocess tools during synthetic pathway development, with which TASY activity was successfully improved by 6.5-fold compared to the highest literature titre in S. cerevisiae cell factories.
Original languageEnglish
Article number200
JournalMicrobial Cell Factories
Volume19
Issue number1
Number of pages12
ISSN1475-2859
DOIs
Publication statusPublished - 2020

Keywords

  • Taxadiene synthase
  • Saccharomyces cerevisiae
  • Paclitaxel, Taxol™
  • Yeast metabolic engineering,
  • Minibioreactor

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