ABSTRACT
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.
Subject(s)
Candida/enzymology , Cytochrome P-450 Enzyme System/immunology , Alkanes , Animals , Cattle , Cross Reactions , Hydroxylation , Immunochemistry , Microsomes/enzymology , Rabbits , Rats , Species SpecificitySubject(s)
Ascomycota/enzymology , Cytochrome P-450 Enzyme System/metabolism , Mixed Function Oxygenases/metabolism , NADPH-Ferrihemoprotein Reductase/metabolism , Saccharomycetales/enzymology , Cytochrome P-450 CYP4A , Intracellular Membranes/enzymology , Kinetics , Microsomes/enzymology , NADPH-Ferrihemoprotein Reductase/isolation & purification , SpectrophotometryABSTRACT
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.
Subject(s)
Candida/metabolism , Cytochrome P-450 Enzyme System/metabolism , NADH, NADPH Oxidoreductases/metabolism , NADPH Dehydrogenase/metabolism , Alkanes , Cytochrome P-450 Enzyme System/isolation & purification , Cytochrome Reductases/analysis , Cytochromes/analysis , Hydroxylation , Mixed Function Oxygenases/analysis , NADPH Dehydrogenase/isolation & purification , NADPH-Ferrihemoprotein Reductase/analysisABSTRACT
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.
