• Biotechnology and Bioprocess Engineering:BBE
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• 제4권1호
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• pp.17-20
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• 1999

Facile synthetic methods of 2',5'-dideoxy-, 2',3'-dideoxy- and 3'-deoxy-1, N6-ethenoadenosine nucleosides by either an enzymatic dideoxyribosyl transfer reaction or a simple chemical reaction were proposed. The synthesis products were isolated and purified by preparative HPLC and their structures were confirmed by 1H NMR (500 MHz) and FAB-MS including high resolution mass measurement. These modified nucleoside analogs have not been reported yet. Therefore, these modified nucleoside analogs are of potential value to be studied further for biological activity such as anticancer or antiviral.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.88-94
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• 2008

Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

• Bulletin of the Korean Chemical Society
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• 제21권3호
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• pp.321-327
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• 2000

Polynucleotide analogues containing pyrimidine (uracil and thynime) bases, $poly[(1'{\beta}-uracil-1-yl-2'.5'-dideoxy-D-glycero-pent-4'-enofuranose)-alt-(maleic$ acid)] (12) and $poly[(1'-{\beta}-thymin-1-yl-2'5'-dideoxy-D-glycero-pent-4'-enofuranose)-alt-(maleic$, acid)] (15), were synthesized by the altermating copolymerization of relevant nucleosied derivatives and maleic anhydride, and the subsequent hydrolysis. The polymers had quite similar structures to the natural polymes and were soluble in water, They showed high hypochromicities up to 49% and excimer fluorescence due to the base stacking, and polyelectrolyte behavior. Since the polymers had compact structrures, depyrimidinations, the release of pyrimidine bases from the polymer backbone, occurred in aqueous solutions with higher rates compared with those of the natural polymers.

• Bulletin of the Korean Chemical Society
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• 제16권9호
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• pp.805-808
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• 1995

Diacetone-D-glucose was converted into 5-azido-6-O-benzoyl-3-O-benzyl-5-deoxy-1,2-O-isopropylidene-α-D-glucofuranose. After removal of isopropylidene and benzoyl protecting groups, hydrogenation performed reduction of azide and subsequent cyclization by reductive amination to give 3-O-benzyl-1-deoxy nojirimycin in high yield. The second azide group was introduced on 2-carbon by selective substitution reaction, and reduction of azide to amino group gave titled compound.

• Bulletin of the Korean Chemical Society
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• 제6권6호
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• pp.350-354
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• 1985

1-[Ethyl (E)-5,6-dideoxy-2,3-O-isopropylidene-$\beta$-D-ribo-hept-5-enofuranosyluronate] uracil(12) was synthesized. Other various heptofuranose uncleosides were also synthesized from uridine and adenosine by two-carbon chain extension using Witting reaction.

• Restorative Dentistry and Endodontics
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• 제23권1호
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• pp.328-338
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• 1998

Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.

• 한국식품영양과학회지
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• 제37권5호
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• pp.612-617
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• 2008

우리나라 남해안 연안에 서식하는 복섬(Takifugu niphobles) 간장의 TTX유도체들을 활성탄 칼럼을 이용하여 부분 정제하고, Hydrophilic Interaction Liquid Chromatography(HILIC)를 사용한 LC/MS(SIM mode)로 분석하였으며, pH와 가열 조건에 따른 독성 변화를 조사하고 복섬 fillet으로 복국 조리시 독성의 분포를 조사하였다. 복섬 간장의 독성분은 LC/MS에 의하여 7개의 성분이 분석되었으며, 각 성분의 함량과 조성은 5,6,11-trideoxyTTX(34.0%, 1,029.6nmol/g), 6,11-dideoxyTTX(29.3%, 887.6 nmol/g), TTX (22.1%, 667.8 nmol/g), 4,9-anhydroTTX(11.2%, 339.3nmol/g), 11-deoxyTTX＋5-deoxyTTX(2.6%, 78.6 nmol/g), 4-epiTTX(0.8%, 23.6 nmol/g), 5,6,11-trideoxyTTX(34.0%), 6,11-dideoxyTTX(29.3%), TTX(22.1%), 4,9-anhydroTTX (11.2%), 4-epiTTX(0.8%)이었다. 복섬 간장 추출물 희석액의 독성은 pH에 따라 크게 변하여 pH 3에서 8.4 MU/mL로 최고의 독성을 나타내었고, 알칼리로 갈수록 독성이 감소하여 pH 10에서는 pH 3의 1/7(1.4 MU/mL)을 나타내었다. 각각의 pH(pH 5, 7, 9)에서 $80^{\circ}C,\;100^{\circ}C,\;115^{\circ}C$를 유지하며 가열 시 온도는 높을수록, 시간은 길수록 독성은 감소하였다. 특히, 산성이나 중성에서는 독성이 완만하게 감소하는 경향을 보였으나, 알칼리 영역인 pH 9에서는 가열 10분 후에 $80^{\circ}C$의 경우라도 최초 독성(79 MU/mL)이 1/2 이하로 급격히 감소하였으며, $115^{\circ}C$에서는 완전 소멸하였다. 복섬 개체별 fillet 중의 총 독량은 $43.2{\sim}106.7$ MU로 개체에 따라서 2.5배까지 독량의 차이를 나타내었으며, 복섬 가식부에 3배량의 물을 가하여 10분간 끓일 경우 총 독량의 $64{\sim}78%$는 국물 중으로 용출되어 나와 육에 남아있는 것보다 독성이 강하였다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.192-192
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• 1994

본 논문에서는 cytochrome P450 LA1 유전자의 5'-upsteam 조절부위의 클로닝을 실시하였다. pUC19 vector에 연결시킨 3.4 Kb 크기의 Pstl DNA조각을 Sst1, Nco1 제한 효소로 자른 뒤, Exonuclease III 를 처리하여 약 200bp 씩의 차이를 갖는 여러 크기의 plasmid들을 얻었다. 이 plasmid 의 핵산서열을 알아보기 위해 dideoxy nucletide를 이용한 sequencing방법으로 그 핵산서열의 결정 실험을 시도하였다. 또한, 다환상 방향족 탄화수소 화합물에 반응성을 갖는 C57BL/6N 생쥐와 반응성을 갖지않는 DBA/2N 생쥐에 있어 phase II 대사 효소인UDP-glucuronosyltransferase 효소활성에 대한 3-methylcholanthrene의 영향을 알아보기 위해 C57BL/6N 생쥐와 DBA/2N 생쥐에 각각 다른 농도의 3-methylcholanthrene을 처리하거나 각기 다른 시간에 3-methylcholanthrene를 처리하였다. 그 결과 UDP-glucuronosyl-transferase의 mRNA가 3-methylcholanthrene양의 증가에 따라, 처치시간이 길어짐에 따라 증가되어지며 그 mRNA위 크기는 약 2.2Kb 정도임을 알았다. 이로부터 UDP-ghucuronosyltransferase 또한 cytochrome P45O와 함께 다환상 방향족 탄화수소 화합물 조절인자를 통한 조절을 받을 것이며 phase I phase II 약물 대사 효소가 조절상 밀접한 관련을 가짐을 예측할 수 있었다.

• 한국양식학회지
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• 제8권3호
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• pp.209-220
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• 1995

미꾸라지의 간으로부터 핵을 분리하여, 저농도 염추출 및 제한효소 처리로 핵기질(nuclear matrix)을 분리하였다. 분리된 핵기질을 Proteinase K로 분해한 후, phenol-chloroform 추출로 크기가 약 0.3kb-15kb의 분포를 나타내는 핵기질 부착 DNA (nuclear matrix attachment regions : MARs)를 얻었다. 효모 URA 3 유전자를 가진 2.13 kb Eco47 III 단편을 제한효소 Ssp I 으로 절단된 pUC19 플라즈미드 벡타에 결합시켜, ARS (autonomously replication sequence) 클로닝을 위한 pURY19 플라즈미드 벡타를 만들었다. 이 pURY19 벡타는 Saccharomyces cerevisiae내에서 독립적으로 복제할 수 없기 때문에, 물고기의 효율적인 발현 벡타 개발을 위해, 이 system을 이용하여, S. cerevisiae내에서 독립적으로 복제 가능한 미꾸라지의 ARS를 클로닝하고자 하였다. 분리 된 MARs를 pURY19 벡타에 결합시 킨 다음, E. coli $DH5\alpha$에 형질전환시켜 $pURY19N_{l-62}$를 얻었다. MAR Libraries $(pURY19N_{1-62})$를 각각 $Ura^-\;S.\;cerevisiae$에 형질전환시켜, S. cerevisiae내에서 독립적으로 복제 가능한 M. mizolepis 유래의 복제원점들 (ARSs)을 분리하여, Sanger's dideoxy-chain termination method로 염기서열을 분석하였다. 염기서열 분석결과 모든 clones들은 AT-rich하였으며, 특히 $pURY19N_6$에는 ARS concensus sequence, Topoisomerase II consensus, near A-box, 그리고 T-box들이 존재하였다.

• Archives of Pharmacal Research
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• 제29권5호
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• pp.375-383
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• 2006

The anti platelet effects of a novel guanidine derivative, KR-32570 ([5-(2-methoxy-5-chlorophenyl) furan-2-ylcarbonyl]guanidine), were investigated with an emphasis on the mechanisms underlying its inhibition of collagen-induced platelet aggregation. KR-32570 significantly inhibited the aggregation of washed rabbit platelets induced by collagen $(10{\mu}g/mL)$, thrombin (0.05 U/mL), arachidonic acid $(100{\mu}M)$, a thromboxane (TX) $A_2$ mimetic agent U46619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin $F_2,\;1{\mu}M$) and a $Ca^{2+}$ ATPase inhibitor thapsigargin $(0.5{\mu}M)$ ($IC_{50}$ values: $13.8{\pm}1.8,\;26.3{\pm}1.2,\;8.5{\pm}0.9,\;4.3{\pm}1.7\;and\;49.8{\pm}1.4{\mu}M$, respectively). KR-32570 inhibited the collagen-induced liberation of $[^3H]$arachidonic acid from the platelets in a concentration dependent manner with complete inhibition being observed at $50{\mu}M$. The $TXA_2$ synthase assay showed that KR-32570 also inhibited the conversion of the substrate $PGH_2$ to $TXB_2$ at all concentrations. Furthermore, KR-32570 significantly inhibited the $[Ca^{2+}]_i$ mobilization induced by collagen at $50{\mu}M$, which is the concentration that completely inhibits platelet aggregation. KR-32570 also decreased the level of collagen $(10{\mu}g/mL)$induced secretion of serotonin from the dense-granule contents of platelets, and inhibited the NHE-1-mediated rabbit platelet swelling induced by intracellular acidification. These results suggest that the antiplatelet activity of KR-32570 against collagen-induced platelet aggregation is mediated mainly by inhibiting the release of arachidonic acid, $TXA_2$ synthase, the mobilization of cytosolic $Ca^{2+}$ and NHE-1.