Long term continuous ATP regeneration by enzymes of the alcohol fermentation pathway and kinases of yeast

Abstract

Continuous ATP regeneration from adenosine using enzymes of the alcohol fermentation pathway, adenosine kinase and adenylate kinase of baker's yeast was investigated using a reactor equipped with a semipermeable membrane. The addition of DTT, which protected the thiol groups of some enzymes against oxidation, increased the duration of the period of ATP formation from 42 h (previously the longest period) to 100 h. With the addition of a yeast extract containing intermediates of alcohol fermentation, a stable steady state was attained in which a yield of more than 75% ATP continued for 2 weeks. These results suggest that the long term continuous ATP formation attained might be due to the protection and stabilization of enzymes by the yeast extract which was added to prevent inactivation.