• Title/Summary/Keyword: Acid site

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Activity of Human Dihydrolipoamide Dehydrogenase Is Largely Reduced by Mutation at Isoleucine-51 to Alanine

  • Kim, Hak-Jung
    • BMB Reports
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    • v.39 no.2
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    • pp.223-227
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    • 2006
  • Dihydrolipoamide dehydrogenase (E3) belongs to the pyridine nucleotide-disulfide oxidoreductase family including glutathione reductase and thioredoxin reductase. It catalyzes the reoxidation of dihydrolipoyl moiety of the acyltransferase components of three $\alpha$-keto acid dehydrogenase complexes and of the hydrogen-carrier protein of the glycine cleavage system. Isoleucine-51 of human E3, located near the active disulfide center Cys residues, is highly conserved in most E3s from several sources. To examine the importance of this highly conserved Ile-51 in human E3 function, it was substituted with Ala using site-directed mutagenesis. The mutant was expressed in Escherichia coli and highly purified using an affinity column. Its E3 activity was decreased about 100-fold, indicating that the conservation of the Ile-51 residue in human E3 was very important to the efficient catalytic function of the enzyme. Its altered spectroscopic properties implied that conformational changes could occur in the mutant.

The penetration site of local anesthetics into liposomal membrane

  • Han, Suk-Kyu;Bae, Song-Ja;Il-Yun;Kim, Nam-Hong
    • Archives of Pharmacal Research
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    • v.8 no.4
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    • pp.205-211
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    • 1985
  • The distribution of local anesthetics between the hydrocarbone interior and surface area of the lipid bilayer of liposomal membrane was calculated employeg fluorescence probe technique. The quenching of fluorescence probe technique. The quenching of fluorescence probe technique. The quenching of fluorescence of 12-(9-anthroyl) stearic acid and N-octadecyl naphthyl-2-amini-6-sulfonic acid by the local anesthetics in liposomal system was used to calculate the distribution. The Stern-Volmer equation was modified and employed for this calculation. The results showed that procaine hydrocloride and benzocaine were mainly distributed on the surface area of the lipid bilayer of the liposoal membrane, while tetracaine hydrochloride penetrated effectively into the hydrocarbon interior and showed even distribution in the lipid bilayer.

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Chemical Compositions of the Observed Precipitation in Forest Area on the Border of Highway(Shingal, Seochun) (고속도로변 산림지역(신갈, 서천) 강우의 화학적 조성)

  • 김영채;정동준;김홍률
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.4 no.4
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    • pp.237-247
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    • 2002
  • Air pollution by acid pollutants is problematic in the whole world. Water acidification has already been deteriorating the forest ecosystem. This study was conducted to analyze the acidity and chemical composition of the open precipitation and throughfall at forests with various geographic locations in Korea. The results of this study are as follows; The open precipitation pH was lowest in Seochun. The throughfall pH showed some buffering capacity in only Quercus mongolica stands. In Pinus rigida(Shingal and Seochun) stands, there was little difference from the open precipitation. Chemical composition of the open precipitation for each sampling site showed that $Ca^{2+}$, N $H_{4}$$^{+}$ and S $O_{4}$$^{2-}$ concentrations had higher value than other ions, and except these ions, the small quantity of ions showed different properties to each site. Changes of ion concentrations in the throughfall showed a tendency to increase. ion concentrations of the throughfall increased with washout and nutrient leaching from the trees. In conclusion, the influence was extended to the pure zone, and the frequency of acid rain is increasing. But, if the deposition of pollutants exceeds the capacity of purification, it would damage forest ecosystem. Further investigations are necessary to identify tolerant tree species to acid pollutants.nts.

Characterization of the Plasmid-Encoded Arsenic Salts Resistance Determinant from Klebsiella oxytoca D12

  • Rhie, Ho-Gun;Lee, Sung-Jae;Lee, Ho-Sa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.593-598
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    • 2004
  • The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K. oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arse) kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced amino acid sequences of the K. oxytoca Dl2 Ars proteins were compared with the amino acid sequences of the E. coli R773 Ars proteins, a significant amino acid similarity was observed (87.9% for ArsR, 89.2% for ArsD, 83.2% for ArsA, 92.6% for ArsB, and 91.3% for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R773 and K. oxytoca Dl2.

Studies on the Developement of Active Components and their Charactrization of 3-Way Catalysts for Autombile Emission Control -Studies on the surface Characterstics Changes of Pd/$\gamma$-Alumina Catalysts by Addition of WO$_3$ and La$_2$O$_3$ as Promoters- (자동차 배기정화용 3원촉매정화기의 국산화 시도를 위한 촉매성분의 개발과 그 촉매 특성에 관한 연구 -WO$_3$ 및 La$_2$O$_3$조촉매성분들의 첨가에 따른 Pd/$\gamma$-Alumina 촉매들의 표면특성 변화-)

  • 이상윤;정석진;박경석
    • Journal of Surface Science and Engineering
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    • v.23 no.2
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    • pp.30-38
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    • 1990
  • For the purpose of suggesting the thermal resistant catalyst for automobile emission control, various catalysts, Pd-WO3 and PD-La2O3 systems, were charactrized before and after thermal aging. It was found that La2O3 formed amorphous surface compound on the support by strong metal-support interaction(SMSI). And by Temperature Programmed Desorption (TPD) expeiment, it was found that the distribution of acid site which is strong acid sites by adding the promoters. After thermal aging, it was observed that the acidity of Pd-WO3 system was decreased largely because of losing acid site by metal vaporization. On the other hand, there was pretty small change in the properties of matter of Pd-La2O3 system. Therefore, it could be considered that La2O3 formed heat resisting amorphous surface compound on the support by SMSI.

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Analysis of the Organic Acid Contaminants on the surface of TEN-STORIED STONE PAGODA ON THE SITE OF WON-GAKSA (원각사지 십층석탑 오염물의 유기산 분석)

  • Lee, Kyu-Shik;Han, Sung-Hee
    • 보존과학연구
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    • s.16
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    • pp.112-122
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    • 1995
  • TEN-STORIED STONE PAGODA ON THE SITE OFWON-GAKSA(Temple) which is one of three marble pagodas in South Korea, were dated from the thirteenth year of the reign of King Sejo(1467). On the roof and surface of each the stories, there were large amount of the contaminants such as pigeon′s excretions, dust and environmental elements for a long time. The pH of contaminants is not acid, but is 7.2, neutral. To find the species of organic acidscontained in the contaminants and the degree of damaging for a marble pagoda, we analyzed the contaminants using GC-MSD method by the following procedures. Organic acids were extracted by saponifying whole contaminants. After Saponification, the organic acids were mathylated to increase their volatility upon subsequent GC-MSD analysis. The mathyl esters of the organic acids are extracted from the acidified aqueous solution. And the organic extracts were washed with adilute base solution. The washed extract were analyzed by GC(Hewlett Packard 5890)with a nonpolar capillary column(Crosslinked 5% Ph Me Silicone, $50×0.2㎜×0.33\mum$film thickness, USA) and Mass Spectrometric Detector(Hewlett Packard 5970B).As the result, it was found that 12 organic acids were the main compound in pagoda′contaminants, and the amount of organic acid were negligible.

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Analysis of Active Center in Hyperthermophilic Cellulase from Pyrococcus horikoshii

  • Kang, Hee-Jin;Ishikawa, Kazuhiko
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1249-1253
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    • 2007
  • A hyperthermostable endoglucanase from Pyrococcus horikoshii with the capability of hydrolyzing crystalline cellulose was analyzed. A protein engineering study was carried out to obtain a reduced-size mutant. Five amino acid residues at both the N- and C-terminus were found to be removable without any loss of activity or thermal stability. Site-directed mutagenesis was also performed on R102, N200, E201, H297, Y299, E342, and W377, residues possibly involved in the active center or in the recognition and binding of a cellulose substrate. The activity of the resulting mutants was considerably decreased, confirming that the mutated residues were all important for activity. A reduced-size enzyme, as active as the wild-type endoglucanase, was successfully obtained, plus the residues critical for its activity and specificity were confirmed. Consequently, an engineered enzyme with a reduced size was obtained, and the amino acids essential for activity were confirmed by site-directed mutagenesis and comparison with a known three-dimensional structure.

Nucleotide Sequence of Pre Protein in Chloramphenicol Resistance Plasmid pKH7. (클로람페니콜 내성 플라스미드 pKH7의 Pre 단백질의 염기서열 결정)

  • 문경호;박봉동;이동석;이백락
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.566-568
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    • 1998
  • Partial nucleotide sequence (nt 1-1842) of chloramphenicol resistance plasmid pKH7 has been reported previously and residual nucleotide sequence (nt 1843-4118) of pKH7 was determined and then the complete nucleotide sequence of pKH7 was obtained. pKH7 consists of 4118 bp and has three ORFs. Besides Rep and CAT proteins described in previous paper, Pre protein which mediates site-specific recombination in Staphylococcus aureus was found to be on pKH7. R $S_{A}$, a site-specific recombination site of Pre protein, and palA, a specific lagging-strand conversion signal, was also found in pKH7. Amino acid sequence of Pre protein of pKH7 was compared with those of other antibiotic resistant Staphylococcus aureus plasmids.s.

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Offsite Risk Assessment on Flammable Hazard Site (가연성물질 저장설비의 사고시 사업장외에 미치는 영향평가)

  • Lee, Dong Hoon;Park, Kyo Shik;Kim, Tae Ok;Shin, Dong Min;Shin, Seo Yun
    • Korean Journal of Hazardous Materials
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    • v.3 no.1
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    • pp.52-58
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    • 2015
  • Since the HF release in 2012 in Korea, it became one of the most significant to evaluate consequence to the vicinity of industry facilities handling hazardous materials. BTX plant is selected to assess off-site risk to check whether the facility satisfies the Chemical Control Law by Korea Government. Accident scenarios were listed using process safety information. The scenarios having effect to the off-site were selected and assessed further according to guideline provided by Korea government. Worst case and alternative scenarios including other interested scenarios were evaluated using ALOHA. Each evaluated scenario was assessed further considering countermeasures. The results showed that the facility handling chloric acid is safe enough and needed no further protections at the moment.