(2014, IF=2.414) Deletion of the HXK2 gene in Saccharomyces cerevisiae enables mixed sugar fermentation of glucose and galactose in oxygen-limited conditions
Deletion of the HXK2 gene in Saccharomyces cerevisiae enables mixed sugar fermentation of glucose and galactose in oxygen-limited conditions
Yi-Hyun Bae; Dae-Hyuk Kweon; Yong-Cheol Park; Jin-Ho Seo
Process Biochemistry 49(4):547-553. 2014.04.01
Abstract
Galactose is one of the major sugar components of red seaweeds along with
glucose. In Saccharomyces cerevisiae, synthesis of galactose-metabolizing enzymes (the Leloir
proteins) is under tight and complex regulation in the presence of glucose by a mechanism
called catabolite repression. As a result, when both sugars are concurrently present in the
medium under oxygen-limited conditions, the yeast cannot utilize galactose even after
glucose consumption. Research efforts on mixed sugar fermentation of glucose/galactose,
therefore, were made in the presence of oxygen, thereby resulting in sequential substrate
consumption, and low ethanol yield and productivity. In this study, mixed sugar fermentation
of glucose and galactose in oxygen-limited conditions was achieved by deleting the HXK2 gene,
a moonlighting protein acting as hexokinase or repressor involved in catabolite repression of
S. cerevisiae. Remarkably, the S. cerevisiae D452-2hxk2 strain utilized galactose after glucose
depletion without a diauxic lag period, whereas the parental strain could not use galactose at
all under oxygen-limited conditions. The hxk2 strain fermented galactose at the consumption
rate of 3.020.10 g/L∙h to produce ethanol with a yield of 0.440.01 g ethanol/g galactose.
Complementation of the hxk2 strain with the plasmid-harbored HXK2 gene under the control
of various promoters clearly showed that the galactose consumption rates were inversely
elated to the mRNA levels of the HXK2 gene. The transcription of the GAL genes were
dramatically elevated by deleting the HXK2 gene, whereas the expressions of hexokinase
genes were not significantly affected. We concluded that the HXK2-deleted strain is able to
efficiently utilize both glucose and galactose in the oxygen-limited conditions by alleviating
catabolite-repression.
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