• 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 the Korean Society of Food Science and Nutrition
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• v.21 no.3
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• pp.279-285
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• 1992

Cytidine deaminase (EC 3.5.4.5) from Aspergillus fumigatus IFO 5840, which was the first cytidine deaminase to be found in a mold, was fractionated with ammonium sulfate (35-60%). When the enzyme solution in 0.25M of Tris-HCI buffer (pH 7.2) was preincubated at $37^{\circ}C$ for 25min, the enzyme activity was reached to maximum state. The optimum pH and temperature for the enzyme activity were found to be 6.8 to 7.2 and near $37^{\circ}C$, respectively. The enzyme was stable in a pH 7.2 to 9.0, and was generally stable at 4$0^{\circ}C$, but after treating at 6$0^{\circ}C$ for 20min at the optimal pH, 17% of the enzyme activity was inactivated, and disappeared completely by treating at 1$0^{\circ}C$ for 25min. Activation energy (Ea) of fungal cytidine deaminase was calculated as 14.190 Kcal /mol by the Arrhenius plot, and temperate coeffient ($Q_{10}$ ) of the enzyme was calculated as 2.163.

• Korean Journal of Environmental Biology
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• v.21 no.1
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• pp.60-65
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• 2003

The cytidine deaminase was partialy purified with sephadex G-200 and the characteristics of the enzyme were clarified. The molecular mass of the plasmid-encoded protein was identified as about 30 kDa in a minicell system. The native enzyme was estimated to have the molecular mass of 60 kDa by gel filtration. This indicates that the native enzyme may exist as a dimer composed of two identical subunits. The enzyme was reasonably stable in the pH range of 6 to 9, and was labile under high temperature above $50^{\circ}C$. Mercaptoethanol, pCMB, mercury and copper were found to inhibit the enzyme activity. The cytidine analogues of bromo- and iodo-(deoxy)-cytidine were also found to inhibit the activity, while fluorodeoxycytidine and azacytidine were found to activate it. Deoxycytidine, cytidine, ara-C and Methyldeoxycytidine have excellent substrates for the enzyme.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.512.2-512
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• 1986

A mutant of bacillus subtilis with a defective cdd gene encoding deoxycytidine-cytidine deaminase(EC 3.5.4.5.) has been characterized genetically. The genetic lesion causing the altered deoxycytidine-cytidine deaminase, cdd, was mapped at 225 min on the linkage map of B.subtilis by AR9 transduction Transductional analysis of the cdd region established the gene order as trp-lys-dnaE-cdd-aroD. The cdd gene was linked 72% with the aroD and 20% with the lys.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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• pp.536-539
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• 1988

A mutant of Bacillus subtilis with a defective cdd gene encoding deoxycytidine-cytidine deaminase (EC 3.5.4.5) has been characterized genetically. The genetic lesion, cdd, causing the altered deoxycytidine-cytidine deaminase was mapped at 225 min on the linkage map of B. subtilis by AR9 transduction, Transductional analysis of the cdd region established the gene order in clockwise as trp-lys-cdd-aroD. The cdd gene was linked 72% with the aroD and 20% with the lys.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.334-342
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• 1989

The Bacillus stearothermophilus cdd gene encoding cytidine deaminase (cytidine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5) was isolated through shot gun cloning by oomplementation of an E. coli cdd mutation. Primarily 3.0 kbp of the exogenote was cloned into the Pstl site of pBR322 (pJSC101). By subsequent deletion and subcloning from the insert of pJSC101 with cdd$^+$ and tetracycline resistancy, about 1.35 kbp of the EcoRI$_1$/PstI$_2$ fragment containing the cdd gene was isolated as pJSC201. The minicell experiment revealed a molecular mass of 33,000 dalton for polypeptide from the cloned DNA fragment complementing the cdd gene. From the lacZ fusion of 550 bp fragment of the EcoRI$_1$/AuaI as a putative promoter region, the transcription direction of the cdd gene on pJSC201 is from EcoRI towards the PstI sites, When the cdd gene was expressed in B. subtilis ED4O (cdd$^-$, pyr$^-$) by transformation with the E. coli-B. subtilis shuttle vector, the gene expression occured more efficiently than in E. coli and the gene appears to be stably maintained in B. subtitis as well as in E. coli.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.640-646
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• 1990

We have cloned the cdd gene from E. coli C600 using (cdd-) as a host. From the sequenced promoter region of E=, coli cdd gene which has been determined by Valentin-Hansen P. (1985), we synthesized the 23 mer oligonucleotides corresponding to the transcription initiation region and used as a probe for cloning of the cdd gene by Southern blotting. The isolated fragments in the blotting were introduced to the colony hybridization after transforming it into the E. coli JF611 (cdd-, pyr double mutant), and we identified the hybridized band at 27 kb long. From the original insert of 27 kb fragment in theBamHI site of pBR322, the 5.3 kb fragment containing the cdd gene was isolated by subsequent deletion and subeloning. From the derived plasmid pTK509, further deletion and subcloning were performed and clarified that the cdd gene was located in the 2.1 kb of SaZI/DraI segment in the insert of pTK605. The polypeptide encoded by the cloned DNA was appeared to be a molecular mass of 33,000.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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• pp.468-475
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• 1988

The cloned B. subtilis cdd gene encoding cytidine/2'deoxycytidine deaminase (EC 3.5.4.5) was expressed in the cdd deficient B. subtilis mutant ED40. The gene was isolted from the cdd complementing plasmid pSO21, and inserted into the EcoR1/Pvu1 sites of pGB215-110 ΔB, which is a temperature sensitivie E. coli-B. subtilis shuttle vector. In the transformed B. subtilis ED4O harboring the resulting plasmid pSO100, cdd was expressed at several hundred fold elevated levels, and the cytidine deaminase activity in E. coli containing pSO100 was twice the level in B. subtilis/pSO0100. The Km value for cytidine of the partially purified enzyme is 1.88$\times$10$^{-4}$M at pH 7.0 and the V$_{max}$ = 11.1 $\mu$mol/min/mg of protein. The enzyme was completely inhibited by 0.1M mercaptoethanol and HgCl$_2$. The inhibition by p-chrolomercurybenzoic acid showed a Ki = 5 uM. These results suggest that sulfhydryl reagents block an active site thiol group, and/or disturb the formation of the tetrameic holoenzyme.

• IMMUNE NETWORK
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• v.12 no.6
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• pp.230-239
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• 2012

Activation-induced cytidine deaminase (AID) is an enzyme that is predominantly expressed in germinal center B cells and plays a pivotal role in immunoglobulin class switch recombination and somatic hypermutation for antibody (Ab) maturation. These two genetic processes endow Abs with protective functions against a multitude of antigens (pathogens) during humoral immune responses. In B cells, AID expression is regulated at the level of either transcriptional activation on AID gene loci or post-transcriptional suppression of AID mRNA. Furthermore, AID stabilization and targeting are determined by post-translational modifications and interactions with other cellular/nuclear factors. On the other hand, aberrant expression of AID causes B cell leukemias and lymphomas, including Burkitt's lymphoma caused by c-myc/IgH translocation. AID is also ectopically expressed in T cells and non-immune cells, and triggers point mutations in relevant DNA loci, resulting in tumorigenesis. Here, I review the recent literatures on the function of AID, regulation of AID expression, stability and targeting in B cells, and AID-related tumor formation.

• Korean Journal of Microbiology
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• v.35 no.2
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• pp.133-138
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• 1999

Essential amino acids involving in the active site ofthe cytidn~e deruninase from Bncillus subtilis ED 213 were determined by chemical modification studies. Tllc purified cytidine deruninase tiom Booillus subtilis ED 213 required the reduced form of Fe(lI)ion. since the enzyme was inhibited 43% by 1 mnM o-phenanthroline. Whereas the enzyme activity was activated up to 28% by 1 1 ethylenediaminetetraacetic acid. The cytidine deaninase activily was completely inhibited by 1 mM N-bromosuccinimide, chloramine-T, and p-chloromercuribenzoic acid (p-CMB), respectively. The enzyme activity was inhibited 36% by 1 mM pyridoxal-S-phosphale, and 31% by 1 mM l-ethy~-3-(3-dirneIhj~laminoprop}~~)c~bodiiamide and glycine inethyl ester. The enzyme activity was strongly inhibited 68% by 1 \mu$M \rho$-CMB and this inhibition of the enzyme activity with 1 \mu$M \rho$-CMB was completely reactivated by 5 mM cysleine as a reducing agent. We speculaled that tyrosine, methionine, cysteuie and/or serine residues are located ui or near ihe active site of the cytidine deruniuase from Bncilus subrilis ED 213 and indirectly related to lysine and/or glycine.