Enzymatic hydrolysis of sphingolipids. 8. Further purification and properties of rat brain ceramidase.
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Biomedical subjects
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Ceramide lactoside [1-O-(galactosido-4-beta-glucosido)-2-N-acyl-sphingosine] was hydrolysed to ceramide glucoside and galactose by beta-galactosidase of rat brain. The reaction was not reversible, required cholate or taurocholate, had optimum pH5.0 and K(m) 2.2x10(-5)m. It was inhibited by gamma-galactonolactone and galactose as well as by ceramide, sphingosine and fatty acid. Ceramide lactoside could be degraded to ceramide, galactose and glucose by mixtures of rat-brain beta-galactosidase and ox-brain beta-glucosidase.
Ceramide glucoside (1-O-glucosido-2-N-acyl-sphingosine) was hydrolysed to ceramide (N-acyl-sphingosine) and glucose by beta-glucosidase from ox brain. The reaction was stimulated by the non-ionic detergent, Triton X-100, or by the anionic detergents, cholate or taurocholate. It was not reversible, had optimum pH5.0 (with acetate buffer) or 5.6 (with pyridine buffer), had K(m) 1.8x10(-4)m and was inhibited by delta-gluconolactone and sphingosine, but not by ceramide or palmitic acid.
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