The enzymes involved in the initial degradative steps of lysine metabolism, lysine-2-oxoglutarate reductase and saccharopine dehydrogenase, were studied and their activities in different mammals compared. Values obtained in human, rat, pig, dog, cat, ox and sheep liver indicated that in vitro, appreciable degradation of lysine to saccharopine (4-6nmol/min per mg of protein) occurred. The specific activity of saccharopine dehydrogenase in most species studied was higher than that of lysine-oxoglutarate reductase. The rate of production of glutamate from saccharopine in each animal species was investigated and related to the rate of production of alpha-aminoadipate. The rate of formation of lysine from saccharopine, catalysed by saccharopine oxidoreductase, was examined and correlated with the dietary intake of lysine in each species studied.

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Lysine-2-oxoglutarate reductase was prepared from ox liver and its characteristics were examined. Its activity was totally inhibited in the presence of NH(4)Cl. Under conditions that inhibit saccharopine formation, and in the presence of NADP(+), ox liver mitochondria were found to catalyse the hydrolysis of saccharopine to lysine and alpha-oxoglutarate. The enzyme involved was named saccharopine oxidoreductase. It was partially purified and separated from lysine-oxoglutarate reductase. Comparison of the properties of these two enzymes showed that saccharopine degradation was stimulated under conditions that inhibit its formation. The effect of pH, various cofactors and stability during incubation confirm that saccharopine biosynthesis from, and degradation to, lysine are catalysed by two distinct enzymes.

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ExtractElevated levels of saccharopine, lysine, and citrulline in urine and plasma were observed in a patient suffering from saccharopinuria. Using radioisotope methods the lysine-degradative enzymes, lysine-oxoglutarate reductase and saccharopine dehyrogenase, were studied in skin fibroblasts grown from this patient and from healthy subjects.The results show that, in contrast to healthy individuals (range 177-320 pmol formed/min/mg protein), the paitent's fibroblasts are completely lacking in saccharopine dehydrogenase activity, which accounts for the presence of the high levels of saccharopine. The patient also has a reduced level of lysine-oxoglutarate reductase activity (333 pmol saccharopine formed/min/mg protein; range 550-1,570 nmol), which may in part explain the hyperlysinemia. A further enzyme saccharopine oxidoreductase which metabolizes saccharopine to lysine was found to be present in the patient's fibroblasts (63 pmol lysine formed/min/mg protein) but absent from those of healthy control subjects. This indicated induction of this enzyme by the patient in an attempt to reduce the high levels of saccharopine in her tissues and body fluid.

Sea lampreys, Mordacia mordax, were collected in spring from the Yarra River, Victoria, during their upstream spawning migration, to study the lipid composition in their tissues and plasma and their lipid transport system. Plasma lipoproteins were isolated by preparative ultracentrifugation and their chemical compositions were analyzed. The major classes of lipoproteins were found to be similar to those of man and higher animals. Lipids from the muscle and liver were analyzed for fatty acids. The striking feature of the lipids in the migrating lamprey is the presence of very high levels of cholesterol in both plasma and muscle. The possible metabolic roles of cholesterol have been discussed.