Phosphagen
kinase systems provide different advantages to tissues with high and fluctuating energy demands, in particular an efficient energy buffering system. In this study we show for the first time functional expression of two phosphagen
kinase systems in Saccharomyces cerevisiae, which does not normally contain such systems. First, to establish the
creatine kinase system, in addition to overexpressing
creatine kinase isoenzymes, we had to install the biosynthesis pathway of
creatine by co-overexpression of
L-arginine:glycine amidinotransferase and
guanidinoacetate methyltransferase. Although we could achieve considerable
creatine kinase activity, together with more than 3 mM intracellular
creatine, this was not sufficient to confer an obvious advantage to the yeast under the specific stress conditions examined here. Second, using
arginine kinase, we successfully installed an intracellular phosphagen pool of about 5 mM
phosphoarginine. Such
arginine kinase-expressing yeast showed improved resistance under two stress challenges that drain cellular energy, which were transient pH reduction and
starvation. Although transient
starvation led to 50% reduced intracellular
ATP concentrations in wild-type yeast,
arginine kinase overexpression stabilized the
ATP pool at the pre-stress level. Thus, our results demonstrate that temporal energy buffering is an intrinsic property of phosphagen
kinases that can be transferred to phylogenetically very distant organisms.