Synthetic and Systems Biotechnology (Jun 2023)

Improving solubility and copy number of taxadiene synthase to enhance the titer of taxadiene in Yarrowia lipolytica

  • Man Xu,
  • Wenliang Xie,
  • Zhen Luo,
  • Chun-Xiu Li,
  • Qiang Hua,
  • Jian-He Xu

Journal volume & issue
Vol. 8, no. 2
pp. 331 – 338

Abstract

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Taxadiene is an important precursor for the biosynthesis of highly effective anticancer drug paclitaxel, but its microbial biosynthesis yield is very low. In this study, we employed Yarrowia lipolytica as a microbial host to produce taxadiene. First, a “push–pull” strategy was adopted to increase taxadiene production by 234%. Then taxadiene synthase was fused with five solubilizing tags respectively, leading a maximum increase of 62.3% in taxadiene production when fused with SUMO. Subsequently, a multi-copy iterative integration method was used to further increase taxadiene titer, achieving the maximum titer of 23.7 mg/L in shake flask culture after three rounds of integration. Finally, the taxadiene titer was increased to 101.4 mg/L by optimization of the fed-batch fermentation conditions. This is the first report of taxadiene biosynthesis accomplished in Y. lipolytica, serving as a good example for the sustainable production of taxadiene and other terpenoids in this oleaginous yeast.

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