Lactobacillus acidophilus, L. bulgaricus, L. casei, L. delbrueckii, L. lactis and L. plantarum contained a pyruvate oxidase for the oxidation of pyruvate to acetyl phosphate and acetate. The presence of an acetate kinase converted the acetyl phosphate to acetate. L. casei and L. plantarum produced lactate and acetoin, in addition to acetate, under the conditions used while L. casei also produced diacetyL L. casei and L. plantarum were the only species to utilize citrate. L. helveticus and L. helveticus subsp. jugurti did not utilize pyruvate under the conditions used.

Pyruvate dehydrogenase activity was detected in whole cells but not in cell-free extracts of Streptococcus lactis. However, the three component enzymes (pyruvate decarboxylase, lipoate acetyltransferase and lipoyl dehydrogenase) of the pyruvate dehydrogenase complex were identified in the cell-free extracts. Whole cells of the three species of group N streptococci formed acetoin and diacetyl only after the pathway forming acetate had become saturated. S. lactis subsp. diacetylactis DRC2 formed more acetoin and diacetyl and less acetate from pyruvate than did S. lactis CW. Strains CIO and DRC2 were able to form acetoin via a-acetolactate or diacetyl and to convert acetoin to butane-2,3-diol. S. cremoris HP was able to form acetoin only via a-acetolactate and could not convert acetoin to butane-2,3cdiol.

The fumarate reductase (NADH) present in cell-free extracts of S. lactis CIO was purified approximately 100-fold by chromatography on DEAE-cellulose in the presence of the non-ionic detergent Teric X-lO, and some of the properties of this partially purified enzyme were characterized. Fumarate was able to act as a terminal electron acceptor and decreased the amount of lactate formed and oxygen used during the metabolism of pyruvate by resting cells of S. lactis. Anaerobic growth of S. lactis on glycerol was not observed and fumarate reduction was not coupled with glycerol-3-phosphate oxidation.

The pathway of formation of acetate from pyruvate in the homofermentative organism L. bulgaricus was studied. Three pathways for the formation of acetate were investigated. These were the formation of acetyl CoA by the pyruvate dehydrogenase (lipoate) system, the formation of acetaldehyde by pyruvate decarboxylase, and the formation of acetyl phosphate by pyruvate oxidase. The first two pathways were eliminated when it was found that the formation of acetate was not inhibited by arsenite and that acetaldehyde was not converted to acetate by L. bulgaricus. The formation of acetyl phosphate and acetate by dialysed cell-free extracts indicated the presence of pyruvate oxidase in L. bulgaricus. The pyruvate oxidase system, unlike the pyruvate dehydrogenase (lipoate) system, was not inhibited by unsaturated fatty acids. The organism was shown to possess both acetate kinase and phosphate acetyltransferase which suggested that acetyl phosphate could be converted to acetate or acetyl CoA.