• Journal of Life Science
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• v.8 no.6
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• pp.673-680
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• 1998

Adenosine deaminase gene from Nocardioides sp. J-326TK was cloned by polymerase chain reaction using primers (PI, PII and PIII) constructed from the highly conserved amino acid sequences among Escherichia coli, mouse and human. A PCR product of about 800bp, as expected from the sequence of E. coli adenosine deaminase gene, was obtained from Nocardioides sp. J-326TK chromosomal DNA double-digested with EcoRI and Pst I. DNA sequencing of the PCR product after cloning into pT7Blue T-vector shows 99.5% and 98.9% homologies in nucleotide and amino acid sequences, respectively, with the E. coli adenosine deaminase whereas 59.5% and 46.8% homologies with the human adenosine deaminase, indicating the evolutionarily relationship of these organisms.

• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.380-383
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• 1996

Adenosine deaminase inhibitor was extracted from culture broth of Streptomyces sp. strain V-8 with ethylacetate. The ethylacetate extract showed the characteristic UV absorption spectrum of actinomycins at 440-450 nm. The ethylacetate extract was compared with respect to inhibitory behavior against adenosine deaminase from calf intestinal mucosa with actinomycin D, -C complex and actinomycin V. The Ki values for actnomycin D, -C complex, and actinomycin V against adenosine deaminase were determined to be 9.9 $\times$ 10$^{-6}$ M, 9.6 $\times$ 10$^{-6}$ M and 9.3 $\times$ 10$^{-6}$ M, respectively. The Ki value for the ethylacetate extract of culture broth against adenosine deaminase was determined to be 5.7 $\times$ 10$^{-6}$ M. The kinetic parameters of actinomycin D, -C complex, -V and ethylacetate extract of culture broth for adenosine deaminase were as follows:I$_{50}$ = 1.5 $\times$ 10$^{-5}$ M (actinomycin D), 2.7 $\times$ 10$^{-5}$ M (actinomycin C complex), 3.5 $\times$ 10$^{-5}$ M (actinomycin V), 8.9 $\times$ 10$^{-6}$ M (ethylacetate extract of culture broth). The adenosine deaminase was inhibited noncompetitively by ethylacetate extract of culture broth as well as by actinomycin D, -C complex and actinomycin V.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.9-14
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• 1997

A bacterium, strain YK 391 producing extracellular cytosine deaminase, has been isolated from soil sample collected near Taegu City and identified. The strain YK 391 was observed to be a motile Gram-negative rod, and did not produced capsule nor spore. The bacterium produced acid from glucose and trehalose, not from arabinose. Esculine was nto hydrolyzed. The isolate could grow anaerobically at 37$\circ $C, but not at 4$\circ $C. Palmitoleic and palmitic acids comprised over 80% of the fatty acid composition of the strain. The strain. The strain YK 391 was identified as Chromobacterium violaceum YK 391 based on its morphological and physiolohical characteristics, and on the fatty acid composition. The extracellular cytosine deaminase produced by Chromobacterium violaceum YK 391 is believed to be unique because it was active not only on cytosine and 5-fluorocytosine but also on cytidine.

• Journal of Life Science
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• v.6 no.2
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• pp.120-128
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• 1996

In the screening of actinomycetes culture filtrate for inhibitor of adenosine deaminase, a novel inhibitor was found in a cultured broth of strain J-144K. The optimum conditions for the adenosine deaminase inhibitor production from the isolated strain J-144K were evaluated. This strain showed the maximum yield of adenosine deaminase inhibitor when grown at pH 7.0 and 30$\circ$C for 60 hours in the medium of 1.0% dextrose, 0.5% yeast extract, 0.5% peptone and 0.1% KH$_{2}$PO$_{4}$ under the aerobic condition. Through the activated charcoal extraction, methanol fractionation, Dowex 50 H$^{+}$ X-8 ion exchange column chromatography, Dowex CI$^{-}$ X-8 ion exchange column chromatography, and Sephadex G-15 gel filtration procedures, this inhibitor was purified with three materials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.1
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• pp.87-93
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• 1999

This study was carried out to investigate the effects of nucleic acid related compounds and metal ions on activities of cytidine deaminase from Bacillus subtilis ED 213. The purified cytidine deaminase was weakly inhibited by 1mM GMP, IMP and ATP, but not affected by other nucleic acid related compounds such as CMP and UDP. The apparent Km values for cytidine, deoxycytidine, 5 methylcy tidine, fluorodeoxycytidine, and 5 bromocytidine were calculated to be 6.6$\times$10-4M, 6.0$\times$10-4M, 0.9$\times$10-4M, 0.8$\times$10-4M, and 2.0$\times$10-3M, respectively. The cytidine deaminase was completely inhibited by 1mM Hg2＋, and mildly inhibited over 40% by metal ions such as Na＋ and Fe2＋. However the enzyme activity was activated more than 40% by 1mM Mg2＋.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.1119-1128
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• 2015

1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl₂ compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

• KSBB Journal
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• v.13 no.6
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• pp.669-674
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• 1998

This study was carried out particularly focusing on he antitumor effect in combination with 5-fluorocytosine(5-FC), antifungal agent, and extracellular cytosine deaminase from Chromobacterium violaceum YK 391 against U-937, K-562 and SNU-C4 cells. While the addition of 10$\mu\textrm{g}$/100 ${\mu}\ell$ of anticancer agent, 5-fluorouracil(5-FU), to U-937, K-562 and SNU-C4 caused the decrease of proliferation 90%, 75% and 93% respectively, the addition of 20 $\mu\textrm{g}$/100 ${\mu}\ell$ of the extracellular cytosine deaminase and 10 $\mu\textrm{g}$/100 ${\mu}\ell$ of antifungal agent 5-FC caused the decrease of proliferation 80%, 70% and 90%, respectively. These results, therefore, reveal that this enzyme has the similar clinical effect for considering of adjuvant antitumor effect. From the above results, the treatment of 5-FC and the cytosine deaminase was very effective and showed the possibility to remove side effects which easily occur by the treatment of 5-FU only. An extracellular cytosine deaminase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.2
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• pp.179-186
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• 1991

The nutritional requirement and cultural condition such as carbon and nitrogen sources, cultural temperature, initial pH, cultural time and aeration for the production of endocellular cytosine deaminase from Aspergillus fumigatus IFO 5840 were investigated. The cultural broth giving maximum cytosine deaminase yield was found to consist of 2% glucose as a carbon source and 1% yeast extract and 0.1% peptone as a nitrogen source. Optimal initial pH of the culture broth was pH 8.5 and the enzyme production in the cell usually reached a maximum after 28 hours of cultivation in the 500ml shaking flask containing 100ml broth at $30^{\circ}C$. The endoenzyme production of the used strain was inhibited by inorganic nitrogen, but activated by orgainc nitrogen, yeast extract.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.169-174
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• 1986

Cytosine deaminase was partialy purified about 10 fold from the ceil-free extract of Aspergillus fumigatus. The partialy purified enzyme was relatively stable in a pH 5.5 to 8.0, but thermo-unstable. The enzyme activity was found in a pH optimum of 7.0 and temperature optimum of 30 to 35$^{\circ}C$. The activation energy calculated to be 13,240 cal/mol. The apparent Michaelis constants Km for cytosine was found to be 1.53 mM and the molecular weight was determined to be approximately 32,000. The enzyme was strongly inhibited by 0.1 mM of Hg$^^{2+}$, Pb$^{2+}$, Cd$^{2+}$ and Fe$^{2+}$, furthermore inhibited by 1mM of ATP, UTP, o-phenanthroline and p-chloromercuribenzoate.

• The Korean Journal of Food And Nutrition
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• v.10 no.1
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• pp.53-59
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• 1997

In this study investigated the effect of cytosine deaminase activity from Aspergillus fumigatus IFO 5840 by cytosine analogues. The results were as follows. The enzyme was strongly inibited by 2-thiouracil, 2-thiocytosine, 6-azacytosine and 2-mercaptopyrimidine. The half inhibitory concentration(HIC) of 2-thiocytosine and 6-azacytosine on cytosine deaminase was 0.80mM and 1.15mM, respectively. The enzyme was inhibited at a certain level by addition of 2-thiocytosine immediately, but was maintained to some extend under the inhibited state by 6-azacytosine in proportion to reaction time. Regardless of kinds of substrate such as cytosine and 5-fluorocytosine, 2-thiocytosine and 6-azacytosine showed action as inhibitors, 2-thiocytosine inhibited cytosine deaminase activity about twice as strong as 6-azacytosine. The enzyme, when cytosine was used as a substrate, was revealed the pattern of competitive inhibition by 2-thiocytosine and 6-azacytosine, The ki value for these compounds was 4.5$\times$10-4M and 1.756$\times$10-3M, respectively. At this point, the Hill coefficient for cytosine, 2-thiocytosine and 6-azacytosine was 1.80, 1.81 and 2.45, respectively.