Pseudomonadsによるphenol性化合物の分解について-1-
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A simple and reliable assay method was devised for determination of the phenol oxygenase activity of <I>Pseudomonas aeruginosa</I>. One loopful of an overnight nutrient agar culture of a test organism was inoculated into 1ml of an assay medium consisting of 0.2% KH2PO4, 0.01% MgSO4⋅7H2O, 0.5% NaCl, 0.05% (NH4)2SO4, and an appropriate concentration of phenol compounds (final pH 7.0). In the case of protocatechuic acid (PA, 0.05%), protocatechualdehyde (PAl, 0.01%), salicylic acid (SA, 0.005%), gentisic acid (Ge, 0.08%), or catechol (Ct, 0.005%), 1ml of 1% FeCl3 solution was dropped into a 24-48 hours' culture of test microbes in the assay medium. A light yellow color appeared as a result positive for phenol oxygenase. When the test compound remained unchanged, a specific colonization took place to exhibit green-purple-black immediately after the addition. In the case of gallic acid (Gl, 0.0025%), pyrogallol (Py, 0.0025%), resorcinol (Re, 0.001%), phloroglucinol (Phl, 0.001%), phenol (Phe, 0.0025%), or o-cresol (Cr, 0.00025%), one or two drops of 2% phosphomolybdic acid solution and 2 to 4 drops of 10% aqueous ammonia were placed into the assay culture. A colorless reaction appeared as a result positive for phenol oxygenase. When the test compound remained unchanged, a blue colorization took place.P. aeruginosa degraded Ge, PA, PAl, and Ct after incubation at 37C for 24-72 hours. P. fluorescens and other pseudomonads degraded PA, PAl, and Ct at 25C for 24 hours, but not at 37C for 72 hours. Ge was degraded by P. aeruginosa, but not by any other pseudomonads at 25C or 37C. Therefore, the degradation of Ge was highly specific to P. aeruginosa.All the tested strains of Enterobacter B and Klebsiella degraded PA and PAl. Most of the Enterobacter B strains and about 50% of the Klebsiella strains degraded Ct. Most of the Enterobcter B strains and three of the Klebsiella strains degraded Ge after incubation at 37C for 72 hours. The other Gram-negative rods tested, however, failed to degrade any of the phenol compounds.
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