E. Boles et al., IDENTIFICATION AND CHARACTERIZATION OF MAE1, THE SACCHAROMYCES-CEREVISIAE STRUCTURAL GENE ENCODING MITOCHONDRIAL MALIC ENZYME, Journal of bacteriology, 180(11), 1998, pp. 2875-2882
Pyruvate, a precursor for several amino acids, can be synthesized from
phosphoenolpyruvate by pyruvate kinase, Nevertheless, pyk1 pyk2 mutan
ts of Saccharomyces cerevisiae devoid of pyruvate kinase activity grew
normally on ethanol in defined media, indicating the presence of an a
lternative route for pyruvate synthesis. A candidate for this role is
malic enzyme, which catalyzes the oxidative decarboxylation of malate
to pyruvate. Disruption of open reading frame YKL029c, which is homolo
gous to malic enzyme genes from other organisms, abolished malic enzym
e activity in extracts of glucose-grown cells. Conversely, overexpress
ion of YXL029c/MAE1 from the MET25 promoter resulted in an up to 33-fo
ld increase of malic enzyme activity. Growth studies with mutants demo
nstrated that presence of either Pyk1p or Mae1p is required for growth
on ethanol, Mutants lacking both enzymes could be rescued by addition
of alanine or pyruvate to ethanol cultures. Disruption of MAE1 alone
did not result in a clear phenotype. Regulation of MAE1 was studied by
determining enzyme activities and MAE1 mRNA levels in wild-type cultu
res and by measuring beta-galactosidase activities in a strain carryin
g a MAE1::lacZ fusion, Both in shake flask cultures and in carbon-limi
ted chemostat cultures, MAE1 was constitutively expressed. A three-to
fourfold induction was observed during anaerobic growth on glucose, Su
bcellular fractionation experiments indicated that malic enzyme in S.
cerevisiae is a mitochondrial enzyme. Its regulation and localization
suggest a role in the provision of intramitochondrial NADPH or pyruvat
e under anaerobic growth conditions. However, since null mutants could
still grow anaerobically, this function is apparently not essential.