Data underlying research on engineering oxygen-independent biotin biosynthesis in Saccharomyces cerevisiae
doi:10.4121/14308007.v1
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doi: 10.4121/14308007
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
CC BY-NC-ND 4.0
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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