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J C Hoogerheide

Publications and source records attributed to J C Hoogerheide.

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Studies on the energy metabolism during the respiratory process by baker's yeast.

Microcalorimetry, in combination with conventional methods for determining metabolic activity, opens the possibility to study the efficiency with which ATP, produced as a result of metabolic activity, is utilized by the cell for energy-requiring synthetic reactions. Using commercial baker's yeast as a test organism and glucose, ethanol, acetic and lactic acids as substrates, the fate of the ATP produced by the respiratory process was studied by measuring oxygen consumption (using the Warburg technique) and the corresponding heat development (using the LKB Flow-Microcalorimeter). From these data heat development per mm3 oxygen consumed was calculated. Values obtained should fall within a heat production range that can be calculated from the combustion heat of the process (maximum heat development) and maximum energy conservation, assuming full participation of ATP in energy-requiring synthetic reactions (minimum heat development). It was found that during the respiratory process of "resting" cells of baker's yeast, regardless of the substrate used, heat development was close to the maximum value inherent with substrate oxidation. Consequently, practically all ATP, produced as a result of the respiratory process, is de-phosphorylated under heat development and thus is not (or very inefficiently) utilized for energy-requiring synthetic reactions. In accordance with this conclusion it was found that addition of 2-4-DNP, a powerful uncoupler of phosphorylation from the respiratory process, did not result in an appriciable increase in heat development. Even in the presence of an assimilable N source, allowing unrestricted growth, initially only a very small percentage of the ATP produced is utilized for synthetic processes. A gradual improvement of this poor economic ATP utilization was observed during the prelogarithmic growth phase. As a possible explanation of this wasteful aerobic metabolism of baker's yeast and its restricted ability to utilize ATP for synthetic processes was mentioned the exceptional low content of messenger RNA, typical for a baker's yeast subjected to a ripening process before harvesting.

Dinitrophenols

Studies on the energy metabolism during anaerobic fermentation of glucose by baker's yeast.

As a result of the intimate association of ADP phosphorylation with alcoholic fermentation, resulting in the synthesis of 2 mole ATP per mole glucose fermented, it may be calculated that a minimum of 672 mucal heat development may be expected for every mm-3 CO2 developed during alcoholic fermentation. When all ATP produced would be fully de-phosphorylated to ADP + Pi (e.g. by ATP-ase activity) a maximum heat development of 1200 mucal per mm-3 CO2 could be expected. Using the LKB-Flow-Microcalorimeter for measurement of heat development and at the same time the Warburg technique for measuring CO2 development during anaerobic glucose fermentation of a baker's yeast suspension, the heat development per mm-3 CO2 produced was calculated over a fermentation period of 90 min. Maintenance of strict anaerobic conditions in the Flow-Microcalorimeter vessel was complicated by diffusion of traces of oxygen via the Teflon transport lines, resulting in excessive heat development values, not representative for the alcoholic fermentation. This problem could be circumvented by removal of traces of oxygen by means of addition of the enzyme glucose-oxidase. Poisoning the respiratory enzyme system of the yeast by addition of KCN or azide, or using respiratory-deficient mutants of the yeast also resulted in heat development values, inherent with alcoholic fermentation. The values obtained were very close to the minimum of 672 mucal per mm-3 CO2, at least during the initial phases of fermentation, indicating that ADP regeneration from ATP, essential for maintaining the high fermentation rate, is not primarily the result of ATP-ase activity, but must be due to participation of ATP in energy-requiring synthetic reactions.

Adenosine Diphosphate