The involvement of nicotinamide adenine nucleotides (NAD � , NADH) in the regulation of glycolysis in Lactococcus lactis was investigated by using 13 C and 31 P NMR to monitor in vivo the kinetics of the pools of NAD �, NADH, ATP, inorganic phosphate (P i), glycolytic intermediates, and end products derived from a pulse of glucose. Nicotinic acid specifically labeled on carbon 5 was synthesized and used in the growth medium as a precursor of pyridine nucleotides to allow for in vivo detection of 13 C-labeled NAD � and NADH. The capacity of L. lactis MG1363 to regenerate NAD � was manipulated either by turning on NADH oxidase activity or by knocking out the gene encoding lactate dehydrogenase (LDH). An LDH � deficient strain was constructed by double crossover. Upon supply of glucose, NAD � was constant an
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