Activation and detection of (Pro)mutagenic chemicals using recombinant strains ofStreptomyces griseus

Two recombinant strains ofStreptomyces griseus have been developed to report on the activation of promutagenic° chemicals. This activation is monitored by reversion of the bacterial test strains to a kana-mycin-resistant phenotype. Strain H69 detects point mutations and was reverted at an increased frequency by acetonitrile, 2-aminoanthracene, 1,2-benzanthracene, benzidine, benzo(a)pyrene, 9,10-dimethyl-1,2-benzanthracene, and glycine. The second strain, FS2, detects frame shift mutations and was reverted at an increased frequency by 1,2-benzanthracene, benzidine, and glycine. Compounds such as butylated hydroxytoluene, catechol, chlorobenzene, hydroquinone, potassium chloride, phenol,cis-stilbene,trans-stilbene, and toluene did not elicit positive responses in either strain. In addition, these strains are capable of detecting direct-acting mutagens such asN-methyl-N’-nitrosoguanidine and ICR-191, providing further evidence of their promise for detecting a wider range of mutagens. To our knowledge, this is the first report of bacterial strains capable of activating promutagenic compounds and detecting their mutagenic metabolites without the benefit of an exogenous activation system such as the rodent liver homogenate (S9).     

PHOTOCHEMICAL OXIDATION OF VINDOLINE and 16-O-ACETYLVINDOLINE

Abstract— Photooxidation of 16-O-acetylvindoline in methanol using chlorpromazine-HCl as a photosensitizing agent produces nearly quantitative yields of an iminium derivative. The 16-O-acetylvindol-ine-iminium product was characterized spectrally by proton NMR and mass spectrometry; and by chemical reduction with sodium borodeuteride. Proton NMR and mass spectral analyses of the reduced product revealed that deuterium was stereospecifically incorporated into the 3-a-position. The iminium product formed by photochemical oxidation is identical to that produced by copper oxidases and peroxidase. Photochemical oxidation provides direct evidence for the involvement of free radicals in Aspidosperma alkaloid transformations.