TY - DATA
T1 - Data underlying research on engineering oxygen-independent biotin biosynthesis in Saccharomyces cerevisiae
PY - 2021/03/26
AU - Jean-Marc Daran
AU - Anna Katarzyna Wrońska
AU - Jack T. Pronk
UR - https://data.4tu.nl/articles/dataset/Data_underlying_research_on_engineering_oxygen-independent_biotin_biosynthesis_in_Saccharomyces_cerevisiae/14308007/1
DO - 10.4121/14308007.v1
KW - Prokariotic pathway
KW - biotin biosynthesis
KW - Vitamin B7
KW - gene dosage
KW - anaerobic growth
KW - Metabolic Engineering Multiple
N2 - An oxygen requirement
for <i>de novo</i> biotin synthesis in <i>Saccharomyces cerevisiae</i> 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 <i>Escherichia coli</i>
biotin-biosynthesis pathway in <i>S.
cerevisiae</i>. Implementation of this pathway required expression of seven <i>E. coli </i>genes involved in fatty-acid
synthesis and three <i>E. coli </i>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 <i>E. coli</i> biotin
pathway. The evolved pathway configuration was reverse engineered in the
diploid industrial <i>S. cerevisiae </i>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 <i>S. cerevisiae</i> and demonstrated its
portability in industrial strain backgrounds.
ER -