Definitive screening accelerates Taxol biosynthetic pathway optimization and scale up in Saccharomyces cerevisiae cell factories.
Saccharomyces cerevisiae
bioprocess optimization
definitive screening design
high-throughput microbioreactor
taxol
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Jan 2022
Jan 2022
Historique:
revised:
08
10
2021
received:
04
08
2021
accepted:
11
10
2021
pubmed:
15
10
2021
medline:
19
1
2022
entrez:
14
10
2021
Statut:
ppublish
Résumé
Recent technological advancements in synthetic and systems biology have enabled the construction of microbial cell factories expressing diverse heterologous pathways in unprecedentedly short time scales. However, the translation of such laboratory scale breakthroughs to industrial bioprocesses remains a major bottleneck. In this study, an accelerated bioprocess development approach was employed to optimize the biosynthetic pathway of the blockbuster chemotherapy drug, Taxol. Statistical design of experiments approaches were coupled with an industrially relevant high-throughput microbioreactor system to optimize production of key Taxol intermediates, Taxadien-5α-ol and Taxadien-5α-yl-acetate, in engineered yeast cell factories. The optimal factor combination was determined via data driven statistical modelling and validated in 1 L bioreactors leading to a 2.1-fold improvement in taxane production compared to a typical defined media. Elucidation and mitigation of nutrient limitation enhanced product titers a further two-fold and titers of the critical Taxol precursors, Taxadien-5α-ol and Taxadien-5α-yl-acetate were improved to 34 and 11 mg L The results of this study highlight the benefits of a holistic design of experiments guided approach to expedite early stage bioprocess development.
Sections du résumé
BACKGROUND
BACKGROUND
Recent technological advancements in synthetic and systems biology have enabled the construction of microbial cell factories expressing diverse heterologous pathways in unprecedentedly short time scales. However, the translation of such laboratory scale breakthroughs to industrial bioprocesses remains a major bottleneck.
METHODS AND MAJOR RESULTS
UNASSIGNED
In this study, an accelerated bioprocess development approach was employed to optimize the biosynthetic pathway of the blockbuster chemotherapy drug, Taxol. Statistical design of experiments approaches were coupled with an industrially relevant high-throughput microbioreactor system to optimize production of key Taxol intermediates, Taxadien-5α-ol and Taxadien-5α-yl-acetate, in engineered yeast cell factories. The optimal factor combination was determined via data driven statistical modelling and validated in 1 L bioreactors leading to a 2.1-fold improvement in taxane production compared to a typical defined media. Elucidation and mitigation of nutrient limitation enhanced product titers a further two-fold and titers of the critical Taxol precursors, Taxadien-5α-ol and Taxadien-5α-yl-acetate were improved to 34 and 11 mg L
CONCLUSIONS
CONCLUSIONS
The results of this study highlight the benefits of a holistic design of experiments guided approach to expedite early stage bioprocess development.
Identifiants
pubmed: 34649302
doi: 10.1002/biot.202100414
doi:
Substances chimiques
Paclitaxel
P88XT4IS4D
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2100414Subventions
Organisme : the Engineering and Physical Sciences Research Council
ID : EP/R513209/1
Organisme : the Royal Society
ID : RSG∖R1∖180345
Organisme : The British Council
ID : 527429894
Organisme : the Novo Nordisk Foundation within the frame of the Fermentation Based Biomanufacturing initiative
ID : NNF17SA0031362
Informations de copyright
© 2021 The Authors. Biotechnology Journal published by Wiley-VCH GmbH.
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