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PURIFICATION AND PROPERTIES OF ENZYMES INVOLVED IN THE PROPIONIC ACID FERMENTATION¹

^a Department of Biochemistry, Western Reserve University, School of Medicine, Cleveland, Ohio

ABSTRACT

ALLEN, S. H. G. (Western Reserve University, Cleveland, Ohio), R. W. KELLERMEYER, R. L. STJERNHOLM, AND HARLAND G. WOOD. Purification and properties of enzymes involved in the propionic acid fermentation. J. Bacteriol. 87:171–187. 1964.—Chromatographic procedures are described for the separation and purification of phosphotransacetylase, acetyl kinase, malic dehydrogenase and coenzyme A (CoA) transferase. Purity of the enzymes was judged by homogeneity in an ultracentrifuge and by specific activity. Phosphotransacetylase was obtained 85% pure with a specific activity of 27.1. The preparation of acetyl kinase was a homogeneous protein with a specific activity of 531. The malic dehydrogenase likewise was homogeneous with a specific activity of 938. The CoA transferase, which was about 56% pure with a specific activity of 42.6, is the purest preparation of this enzyme yet described. The pH optimum was 6.5 to 7.8, and the K_m for succinyl-CoA in the transfer of CoA to acetate was found to be 1.3 x 10^–4M; for acetate, in the same transfer, the K_m was 7.0 x 10^–3M; for succinyl-CoA to propionate it was 6.8 x 10^–5M, and for propionate, in the same reaction, 6.2 x 10^–4M. Methods are described for the enzymatic production of methyl-malonyl-CoA, malonyl-CoA, propionyl-CoA, acetyl-CoA, and succinyl-CoA. The role of these enzymes in the propionic acid fermentation as well as the possible mechanism responsible for the high yields of adenosine triphosphate from glucose are considered.

² Present address: Department of Biochemistry, Albany Medical College of Union University, Albany, N.Y.

¹ Presented in part at the 47th meeting of the Federation of American Societies for Experimental Biology, Atlantic City, N.J., April 1963 (Allen et al., 1963b).

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