Construction of ajmalicine and sanguinarine de novo biosynthetic pathways using stable integration sites in yeast

Yeast cell factories have been increasingly employed for producing plant‐derived natural products. Unfortunately, the stability of plant natural product biosynthetic pathway genes, particularly when driven by the same sets of promoters and terminators, remains one of the biggest concerns for synthet...

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Veröffentlicht in:Biotechnology and bioengineering 2022-05, Vol.119 (5), p.1314-1326
Hauptverfasser: Liu, Tengfei, Gou, Yuanwei, Zhang, Bei, Gao, Rui, Dong, Chang, Qi, Mingming, Jiang, Lihong, Ding, Xuanwei, Li, Chun, Lian, Jiazhang
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Sprache:eng
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Zusammenfassung:Yeast cell factories have been increasingly employed for producing plant‐derived natural products. Unfortunately, the stability of plant natural product biosynthetic pathway genes, particularly when driven by the same sets of promoters and terminators, remains one of the biggest concerns for synthetic biology. Here we profile genomic loci flanked by essential genes as stable integration sites in a genome‐wide manner, for stable maintenance of multigene biosynthetic pathways in yeast. We demonstrate the application of our yeast integration platform in the construction of sanguinarine (24 expression cassettes) and ajmalicine (29 expression cassettes) de novo biosynthetic pathways for the first time. Moreover, we establish stable yeast cell factories that can produce 119.2 mg L−1 heteroyohimbine alkaloids (containing 61.4 mg L−1 ajmalicine) in shake flasks, representing the highest titer of monoterpene indole alkaloids (MIAs) ever reported and promising the complete biosynthesis of other high‐value MIAs (such as vinblastine) for biotechnological applications. Stable integration sites flanked by two essential genes were profiled in a genome‐wide manner in Saccharomyce scerevisiae. Using the yeast integration platform, the authors reported for the first time on the construction of yeast cell factories for de novo biosynthesis of sanguinarine (a representative benzylisoquinoline alkaloid) and ajmalicine (a monoterpene indole alkaloid). Although ~30 expression cassettes driven by the same sets of promoters and terminators were integrated, the heterologous multigene biosynthetic pathways could be stably maintained in yeast.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.28040