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화학적수식에 의한 Bacillus subtilis ED 213 Cytidine Deaminase의 활성부위에 관한 연구
박정문,박상원,서태수,김정,유대식,Park, Jung-Moon,Park, Sang-Won,Suh, Tae-Soo,Kim, Jung,Yu, Tae-Shick 한국미생물학회 1999 미생물학회지 Vol.35 No.2
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. Bacillus subtilis ED 213의 cytidine deaminase 의 활성부위에 존재하는 필수 아미노산잔기를 화학수식 방법으로 측정하였다. 본 효소는 1mM o-phenanthroline 에 의하여 효소활성이 43% 저해되어 효소활성 발현에 Fe\sup 2+\가 요구된다고 추정되며, 1mM ethylenediaminetetraacetic acid 에 의해서는 효소활성이 오히려 28% 정도 촉진되었다. 본 효소는 1mM N-bromosuccinimide, 1mM chloramine-T 와 1mM $\rho$-chloromercuribenzoic acid에 의하여 100% 저해되었으며, 그의 저해 양상은 경쟁적 저해 양상을 나타내었다. 본 효소의 효소활성은 1mM pyridoxal-5-phosphate 에 의항 36% 저해되었으며, 1mM 1ethyl-3-carbodiamide 와 1mM glycine methylester에 의해 저해된 효소활성이 5mM cysteine에 의해 완전히 회복되었다. 이상의 결과로부터 Bacillus subtilis ED 213 cytidine deaminase의 활성부위에는 tyrosine, methionine, cysteine 과 serine 잔기가 관여할 뿐만 아니라 lysine 과 glycine 도 효소활성에 관여하는 것으로 추정된다.
Bacillus subtilis ED 213 Cytidine Deaminase 활성에 미치는 핵산관련물질의 영향
박정문(Jung-Moon Park),유대식(Tae-Shick Yu),서태수(Tae-Soo Suh),김정배(Jung-Bae Kim),윤종국(Chong-Guk Yoon) 한국식품영양과학회 1999 한국식품영양과학회지 Vol.28 No.1
Bacillus subtilis ED 213을 lysine, casamino acid와 cytidine과 kanamycine을 첨가한 Spizizen 최소배지 에서 30℃에서 24시간 진탕배양하여, 집균한 균체를 초음파 파쇄시켜 조효소액을 조제후, 조효소액으로부터 cytidine deaminase를 정제하였다. 정제된 효소의 활성은 핵산계 조미료의 정미 성분인 1mM GTP, IMP와 ATP에 의하여 약하게 저해되며, cytidine, deoxycytidine, 5-methycytidine, fluorodeoxycytidine과 5-bromocytidine에 대한 Km값은 각각 6.6×10^(-4)M, 6.0×10^(-4)M, 0.9×10^(-4)M, 0.8×10^(-4)M, 2.0 ×10^(-3)M로 계산되었다. 이 효소의 활성은 1mM Hg^(2+)에 의하여 완전히 저해되었으며, Na^+와 Fe^(2+)에 의해서는 약하게 저해되었다. 그러나 1mM Mg^(2+)에 의하여 이 효소의 활성은 약 40% 이상 촉진되었으며 10mM Mg^(2+)에 의하여 최고값의 효소 활성의 촉진을 나타냈다. 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-methylcytidine, fluorodeoxycytidine, and 5-bromocytidine were calculated to be 6.6×10^(-4)M, 6.0×10^(-4)M, 0.9×10^(-4)M, 0.8×10^(-4)M, and 2.0×10^(-3)M, respectively. The cytidine deaminase was completely inhibited by 1mM Hg^(2+), and mildly inhibited over 40% by metal ions such as Na^+ and Fe^(2+). However the enzyme activity was activated more than 40% by 1mM Mg^(2+).
Leptospira canicola 실험적 감염햄스타의 조직절편내 면역조직화학적 균체항원검출
구복경,김순복,서정향,문윤경,정석찬,박정문,박용호,문진산,Ku, Bok-gyeong,Kim, Soon-bok,Sur, Jung-hang,Moon, Oun-kyong,Jung, Suk-chan,Park, Jung-moon,Park, Yong-ho,Mun, Jin-san 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.2
The present study was done to detect Leptospira canicola antigens in paraffin sections from experimentally infected hamsters with PAP method. The anti-Leptospira canicola serum used as first antibody was prepared by immunizing rabbits with Leptospira canicola. Positive reactions were detected in interstitium, tubular epithelial cells and glomerulus of the kidney, and in the hepatic sinusoid. With PAP method, leptospiral antigens were detected at the 48 hours after bacterial infection, but isolation, silver stain, and dark field microscopic examination of the samples did 48 hours later than PAP method. The results suggested that PAP method is considered as a reliable tool for confirmative diagnosis of this disease.
국내 돼지의 Streptococcus suis 감염율 조사 및 혈청형 동정
정병열,정석찬,김봉환,박용호,박정문,Jung, Byeong-yeal,Jung, Suk-chan,Kim, Bong-hwan,Park, Yong-ho,Park, Jeung-moon 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.3
To investigate the prevalence of Streptococcus suis in pig industry in Korea, we isolated S suis from pigs during two years. The isolates were identified by biochemical and coagglutination test. Serotypes and antimicrobial susceptibility of the isolates were determined. Fourteen strains(12.2%), 11 strains(27.5%), 58 strains(7.8%) and 4 strains(11.1%) of S suis were isolated from 115 nasal swabs, 40 tonsils of healthy pigs, 745 pneumonic lungs and 36 meningitis of diseased pigs, respectively. The isolates were highly susceptible to ampicillin, cephlothin and ofloxacin while they were resistant to oxytetracycline. Among the isolates, 63(75.9%) strains were to the S suis capsular type 1 to 10 and 11(13.3%) strains were untypable. Capsular type 2 was the most prevalent with 32(38.6%) strains of all isolates.