RESUMO
The immunological relations of the cytochrome P-450 from the n-alkane utilizing yeast Candida maltosa to cytochrome P-450 forms of other organisms - yeasts, bacteria and mammalia - were investigated using a solid-phase double-antibody radioimmunoassay. Only the microsomal fraction of other n-alkane utilizing yeasts shows a distinct cross-reaction with an antiserum against cytochrome P-450 from Candida maltosa. Neither the tested bacterial nor the mammalian cytochromes P-450 cross-react with the antiserum.
Assuntos
Candida/enzimologia , Sistema Enzimático do Citocromo P-450/imunologia , Alcanos , Animais , Bovinos , Reações Cruzadas , Hidroxilação , Imunoquímica , Microssomos/enzimologia , Coelhos , Ratos , Especificidade da EspécieAssuntos
Ascomicetos/enzimologia , Sistema Enzimático do Citocromo P-450/metabolismo , Oxigenases de Função Mista/metabolismo , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Saccharomycetales/enzimologia , Citocromo P-450 CYP4A , Membranas Intracelulares/enzimologia , Cinética , Microssomos/enzimologia , NADPH-Ferri-Hemoproteína Redutase/isolamento & purificação , EspectrofotometriaRESUMO
The growth of the investigated Candida guilliermondii strain on n-alkanes induces an alkane-hydroxylating enzyme system, which consists of a cytochrome P-450 and a NADPH-dependent reductase. The cytochrome P-450 was purified to 4 nmoles per mg protein. Long-chain alkanes, preferably hexadecane to octadecane, are hydroxylated to the corresponding primary alcohol by this enzyme system. The substrate induces a type I spectrum, other compounds checked type II spectra.
Assuntos
Candida/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , NADH NADPH Oxirredutases/metabolismo , NADPH Desidrogenase/metabolismo , Alcanos , Sistema Enzimático do Citocromo P-450/isolamento & purificação , Redutases do Citocromo/análise , Citocromos/análise , Hidroxilação , Oxigenases de Função Mista/análise , NADPH Desidrogenase/isolamento & purificação , NADPH-Ferri-Hemoproteína Redutase/análiseRESUMO
In the investigated Candida guilliermondii strain after growth on n-alkanes as the only carbon and energy source 5--10 nMol cytochrome P-450 per g cells (wet weight) could be detected. Cytochrome P-450 and alkane hydroxylase activity was found in the 100 000 xg pellet. Cofactor studies and inhibition experiments revealed the existence of a NADPH-dependent cytochrome P-450 alkane hydroxylase system.
