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Intracellular pH and the role of D-lactate dehydrogenase in the production of metabolic end products by Leuconostoc lactis

Published online by Cambridge University Press:  01 June 2009

Richard J. Fitzgerald ,
Shawn Doonan ,
Larry L. McKay  and
Timothy M. Cogan
Richard J. Fitzgerald
Affiliation:
National Dairy Products Research Centre, Moorepark, Fermoy, Co. Cork, Irish Republic Department of Biochemistry, University College, Cork, Irish Republic
Shawn Doonan
Affiliation:
Department of Biochemistry, University College, Cork, Irish Republic
Larry L. McKay
Affiliation:
Department of Food Science and Nutrition, University of Minnesota, St Paul, MN 55108, USA
Timothy M. Cogan
Affiliation:
National Dairy Products Research Centre, Moorepark, Fermoy, Co. Cork, Irish Republic
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Summary

The kinetics of lactate dehydrogenase from Leuconostoc lactis NCW1 were studied. The pH optimum for the enzyme depended on the concentration of pyruvate used in the assay and the enzyme displayed an ordered mechanism with respect to substrate binding. The Km for pyruvate and NADH and the Vmax of the enzyme decreased 20–, 30– and 6-fold respectively as the pH decreased from 8·0 to 5·0. No activators were found and none of the intermediates of the phosphoketolase pathway tested inhibited the enzyme. ATP, ADP, GTP and NAD+ were inhibitory. The intracellular volume (Volin) and intracellular pH (pHin) decreased as the extracellular pH (pHex) decreased. Co-metabolism of citrate and glucose affected the Volin but did not affect the pHin, which decreased by 0·6 units per unit change in pHex; at pH 7·0, the pHin and pHex were equal. The results suggest that pHin may play a role in determining the production of diacetyl and acetoin at low pH by Leuconostoc.

Type
Original Articles
Information
Journal of Dairy Research , Volume 59 , Issue 3 , August 1992 , pp. 359 - 367
Copyright
Copyright © Proprietors of Journal of Dairy Research 1992

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