• Korean Journal of Microbiology
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• v.25 no.3
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• pp.212-220
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• 1987

The taxonomical properties of strain J-275L isolated from soil as a microorganism which produces extracellular adenine deaminase and cultural conditions for the enxyme production were studied. The hyphae of strain J-275L is fragmented into rod-or coccus-like elements. The elements of fragmented aerkal hyphae has smooth surfaces. The cell wall of the organism contains LL-diaminopimelic acid. Mycolic acid are not produced. As a result of taxonomical studies, strain J-275L is designated as Nocardioides sp. J-275L. The optimum medium for the enzyme production from Nocardioides sp.J-275L wascomposed of 0.5% peptone, 0.5% dextrin, 1% yeast extract, and 0.2% $K_{2}HPO_{4}$. The optimum initial pH of the medium was pH 7.5.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.306-311
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• 1987

After series of purification by means of ammonium sulfate fractionation, the 1st and 2nd DEAE-Cellulose, DEAE-Sephadex A-50, and Sephacryl S-200 superfine gel filtration, the activity of extracellular adenine deaminase from Streptomyces sp. J-350P increased 1764 fold and the yield was 0.3% of original activity. The enzyme was stable at the pH range 6.5 to 8.5 and at up to 5$0^{\circ}C$. The optimum pH and temperature of the enzyme were around 6.5 and 35$^{\circ}C$. The molecular weight ol the enzyme was estimated as 36, 000 by calibrated Sephacryl S-200 superfine column chromatography.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.312-318
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• 1987

The apparent Michaelis constant Km of extracellular adenine deaminase from Streptomyces sp. J-350P was 5.8$\times$10$^{-5}$M. The activation energy or the enzyme was calculated from Arrhenius plots for adenine and the value was 3.13 Kcal/mole. The purine analogues, 6-chloropurine, 2,6-diaminopurine, 6-bromopurine, 4-aminopyrazolo[3,4-d] pyrimidine, 6-iodopurine, and 8-bromoadenine were substrates for the enzyme. 6-Dimethylaminopurine was a competitive inhibitor of the enzyme. The enzyme was inhibited by 0.1mM of Fe$^{3+}$, Ag+, and Hg$^{2+}$ and 1 mM of $\alpha$, $\alpha$＇-dipyridyl, Penta-chiorophenol, and p-chloromercuribenzoate.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.221-228
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• 1987

The extracellular adenine deaminase from Nocardioides sp. J-275L was purified by the following techniques: ammonium sulfate fractionation, DEAE-Cellulose, DEAE-Sephadex A-50 column chromatography, and Sephacryl S-200 superfine gel filtration. The enzyme was partially purified about 3889.5-fold with about 5.2% yield by these procedures. The molecular weight of the enzyme was 39,000 by a calibrated Sephacryl S-200 superfine column chromatography. The enzyme was stable at pH 7.5 and up to $40^{\circ}C$. Glycerol was effective on the stabilization of the enzyme during storage. The optimum pH and temperature of the enzyme were around pH 7.5 and $40^{\circ}C$, respectively. The apparent Michaelis constant Km of the enzyme for adenine was $7.4\times 10^{-5}$M. The purine analogues, 6-chloropurine, 2,6-diaminopurine, 6-bromopurine, 4-aminopyrazolo [3.4-d]pyrimidine, and 8-azaadenine were substrates for the enzyme. 6-Dimethylaminopurine was a competitive inhibitor of the enzyme. The enzyme was inhibited by 1mM of $Cu^{2+}, Fe^{3+}, Pb^{2+}, Hg^{2+}$, and $Ag^{+}$, and 1mM of $\alpha$,$\alpha$'-dipyridyl, pentachlorophenol, and pCMB.