Data underlying research on engineering oxygen-independent biotin biosynthesis in Saccharomyces cerevisiae

doi: 10.4121/14308007.v1
The doi above is for this specific version of this dataset, which is currently the latest. Newer versions may be published in the future. For a link that will always point to the latest version, please use
doi: 10.4121/14308007
Datacite citation style:
Jean-Marc Daran; Wrońska, Anna Katarzyna; Pronk, Jack T. (2021): Data underlying research on engineering oxygen-independent biotin biosynthesis in Saccharomyces cerevisiae. Version 1. 4TU.ResearchData. dataset. https://doi.org/10.4121/14308007.v1
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Dataset
An oxygen requirement for de novo biotin synthesis in Saccharomyces cerevisiae precludes the application of biotin-prototrophic strains in anaerobic processes that use biotin-free media. To overcome this issue, this study explores introduction of the oxygen-independent Escherichia coli biotin-biosynthesis pathway in S. cerevisiae. Implementation of this pathway required expression of seven E. coli genes involved in fatty-acid synthesis and three E. coli genes essential for the formation of a pimelate thioester, a key precursor of biotin synthesis. A yeast strain expressing these genes readily grew in biotin-free medium, irrespective of the presence of oxygen. However, the engineered strain exhibited lower specific growth rates in biotin-free media than in biotin-supplemented media. Following adaptive laboratory evolution in anaerobic cultures, evolved cell lines that no longer showed this growth difference were characterized by genome sequencing and proteome analyses. The evolved isolates exhibited several genomic rearrangements, including a whole-genome duplication, which caused alterations in the relative gene dosages of biosynthetic pathway genes. These alterations resulted in a reduced abundance of the enzymes catalyzing the first three steps of the E. coli biotin pathway. The evolved pathway configuration was reverse engineered in the diploid industrial S. cerevisiae strain Ethanol Red. The resulting strain grew at nearly the same rate in biotin-supplemented and biotin-free media. This study established the first genetic engineering strategy to enable biotin-independent anaerobic growth of S. cerevisiae and demonstrated its portability in industrial strain backgrounds.
history
  • 2021-03-26 first online, published, posted
publisher
4TU.ResearchData
funding
  • Predictive and Accelerated Metabolic Engineering Network (grant code 722287) [more info...] European Commission
organizations
TU Delft, Faculty of Applied Sciences, Department of Biotechnology

DATA

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