||Gene Engineering in Yeast for Biodegradation: Immunological Cross-Reactivity among Cytochrome P-450 System Proteins of 'Saccharomyces cerevisiae' and 'Candida tropicalis'.
Loper, J. C. ;
Chen, C. ;
Dey, C. R. ;
||Cincinnati Univ. Medical Center, OH.;Environmental Protection Agency, Cincinnati, OH. Risk Reduction Engineering Lab.
Genetic engineering ;
Cross reactions ;
Cytochrome P-450 ;
Saccharomyces cerevisiae ;
Hazardous wastes ;
Immune sera ;
Aromatic polycyclic hydrocarbons ;
Polyacrylamide gel electrophoresis ;
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Yeasts are eukaryotic microorganisms whose cytochrome P-450 monooxygenase systems may be amenable to genetic engineering for the hydroxylation and detoxication of polychlorinated aromatic hydrocarbons. The molecular genetic properties of strains of bakers yeast, Saccharomyces cerevisiae, and an n-alkane utilizing yeast, Candida tropicalis ATCC750 are examined. Standard methods were used to purify cytochrome P-450 and NADPH-cytochrome c (P-450) reductase proteins from cells cultured by semi-anaerobic glucose fermentation (S. cerevisiae, C. tropicalis) and by growth on tetradecane (C. tropicalis). Polyvalent antisera prepared in rabbits to some of these proteins were used in tests of immunological relatedness among the purified proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose filter immunoblots. The results provide evidence for gene relationships which should prove useful in gene isolation and subsequent engineering of P-450 enzyme systems in yeast.