• BMB Reports
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• 제37권3호
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• pp.330-338
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• 2004

Different types of cardiotoxin (I-V and n) were isolated and purified from the venom of the Taiwan cobra (Naja naja atra). The effects of these cardiotoxins were studied on membrane-bound acetylcholinesterase, which was isolated from a sheep's brain cortex. The results showed that cardiotoxins I-III, V, and n activated the enzyme by modification of substrate inhibition, but cardiotoxin IV's reaction was different. The inhibition and activation of acetylcholinesterase were linked to the functions of the hydrophobicity index, presence of a cationic cluster, and the accessible arginine residue. Our results indicate that Cardiotoxins have neither a cationic cluster nor an arginine residue in their surface area of loop I; therefore, in contrast to fasciculin, cardiotoxins are attached by loop II to the peripheral site of the enzyme. As a result, fasciculin seems to stabilize nonfunctional conformation, but cardiotoxins seem to stabilize the functional conformation of the enzyme. Based on our experimental and theoretical findings, similar secondary and tertiary structures of cardiotoxins and fasciculin seem to have an opposite function once they interact with acetylcholinesterase.

• 대한환경공학회지
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• 제22권2호
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• pp.375-383
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• 2000

유일로 오염된 지역의 토양에서 toluene을 탄소원으로 이용하는 혼합미생물을 분리하여 toluene, benzene, ethylbenzene 및 xylene isomers(BTEX)의 분해특성을 관찰하였다. 단일기질 실험에서는 모든 BTEX의 분해가 이루어졌으며 toluene, benzene, ethylbenzene, p-xylene 순서로 분해되었다. BTEX 혼합기질 분해실험에서는 단일기질일 때보다 분해속도가 상대적으로 느려졌으며, ethylbenzene이 benzene보다 먼저 분해되는 것이 관찰되었다. 이중 혼합물질 반응 실험에서는 방해작용(inhibition), 촉진작용(stimulation), 그리고 비반응(non-interaction)과 같은 다양한 기질반응이 관찰되었으며, ethylbenzene은 benzene, toluene, xylene의 분해에 강한 방해영향을 주었다. Xylene 분해특성에서 m- 및 p-xylene은 혼합미생물에 탄소원으로 이용되었으며 benzene이나 toluene이 동시에 존재할 때는 xylene isomer의 분해가 촉진되었다. 그러나 o-xylene의 분해는 benzene에 의해서만 촉진되었다.

• Applied Biological Chemistry
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• 제40권4호
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• pp.277-282
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• 1997

아라비돕시스 탈리아나의 아세토락테이트 합성 효소 (ALS)를 그 유전자를 포함하고 있는 대장균 MF 2000/pTATX로부터 부분 정제하였다. 부분 정제된 이 효소를 가지고 여러 가지 변형 화학물질들 즉, 요오드아세트산, 요오드아세타마이드, N-에틸말레이미드 (NEM), 5,5'-디티오비스(2-니트로벤조산) (DTNB), 파라염화수은벤조산 (PCMB), 그리고 페닐글리옥살 등에 대한 민감성을 조사하였다. PCMB가 가장 민감하게 저해를 했으며, DTNB와 NEM이 그 뒤를 따랐다. 이 효소의 기질인 피루브산이 요오드아세트산에 의한 활성 저해를 보호하지 못하였으므로 기질의 결합에 시스테인의 관련이 없는 것 같이 보인다. 한편, 기질이 페닐글리옥살에 의한 효소의 활성 저해를 부분적으로 보호하는 것으로 보아 기질이 아르기닌기와 상호 작용함을 암시하고 있다. 부분 정제된 효소는 발린과 이소루신에 민감하게 방해를 받았으나 루신은 그렇지 않았다. 그러나, PCMB로 변형시킨 효소는 되먹임 방해를 더 강하게 받았다. 그 외 ALS에 대한 새로운 제초제 후보인 피리미디설퍼 벤조산 유도체의 저해 효과를 검토하였다.

• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.909-915
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• 2002

The biodegradation of BTEX components (benzene, toluene, ethylbenzene, o-xylene, m-xylene, and p-xylene) individually and in mixtures was investigated using the o-xylene-degrading thermo-tolerant bacterium Ralsronia sp. strain PHS1 , which utilizes benzene, toluene, ethylbenzene, or o-xylene as its sole carbon source. The results showed that as a single substrate for growth, benzene was superior to both toluene and ethylbenzene. While growth inhibition was severe at higher o-xylene concentrations, no inhibition was observed (up to 100 mg $l^-1$) with ethylbenzene. In mixtures of BTEX compounds, the PHS1 culture was shown to degrade all six BTEX components and the degradation rates were in the order of benzene, toluene, o-xylene, ethylbenzene, and m- and p-xylene. m-Xylene and p-xylene were found to be co-metabolized by this microorganism in the presence of the growth-supporting BTEX compounds. In binary mixtures containing the growth substrates (benzene, toluene, ethylbenzene. and o-xylene), PHS1 degraded each BTEX compound faster when it was alone than when it was a component of a BTEX mixture, although the degree of inhibition varied according to the substrates in the mixtures. p-Xylene was shown to be the most potent inhibitor of BTEX biodegradation in binary mixtures. On the other hand, the degradation rates of the non-growth substrates (m-xylene and p-xylene) were significantly enhanced by the addition of growth substrates. The substrate utilization patterns between PHS1 and other microorganisms were also examined.

• Bulletin of the Korean Chemical Society
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• 제28권5호
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• pp.758-762
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• 2007

A new alkyldithiocarbonate (xanthate), as sodium salts, C2H5OCS2Na, was synthesized by the reaction between CS2 with ethyl alcohol in the presence of NaOH. The new xanthate was characterized by 1H NMR, IR and elemental analysis. Then, the new synthesized compound was examined for functional study of cresolase activity of Mushroom Tyrosinase (MT) from a commercial source of Agricus bisporus in 10 mM phosphate buffer pH 6.8, at three temperatures of 10, 20 and 33℃ using UV spectrophotemetry. 4-[(4-methylphenyl)- azo]-phenol (MePAPh) was used as a synthetic substrate for the enzyme for cresolase reaction. The results show that ethyl xanthate can activate or inhibit the cresolase activity of mushroom tyrosinase depending to the concentration of ethyl xanthate. It was concluded that the enzyme has two distinct sites for ethyl xanthate. The first one is a high-affinity activation site and the other is a low-affinity inhibition site. Activation of the enzyme in the low concentration of ethyl xanthate arises from increasing the affinity of binding for the substrate as well as increasing the enzyme catalytic constant. The affinity of ligand binding in the activation site is decreased by increasing of the temperature, which is the opposite result for the inhibition site. Hence, the nature of the interaction of ethyl xanthate is different in two distinct sites. The binding process for cresolase inhibition is only entropy driven, meanwhile the binding process for cresolase activation is not only entropy driven but also enthalpy driven means that hydrophobic interaction is more important in the inhibition site.

• 유기물자원화
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• 제21권1호
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• pp.62-68
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• 2013

회분식 실험을 통해 매몰지 내 유기물의 농도가 분해 속도에 미치는 영향을 평가하였다. 기질은 돈 및 우육을 이용하였으며 기질의 농도는 2, 4, 6, 8 및 10 g VS/L로 선정하였다. 기질의 농도가 2 g VS/L 일 경우에 돈 및 우육의 메탄 발생율 각각 46.3 및 48.4 ml CH4/g VS.d 로 가장 높게 나타났으며 기질의 농도가 증가할수록 메탄 발생율은 감소하였다. 비선형 저해 방정식을 이용하여 평가된 저해 상수 값은 돈육의 경우, n 및 m은 각각 4.9 및 0.6으로 나타났으며 우육은 각각 1.1 및 0.4로 나타났다(n: 최대 메탄 발생율 저해 상수, m: 최종 메탄 수율 저해 상수). 기질의 농도가 증가할수록 메탄 발생율은 민감하게 반응하였으나 최종 메탄 수율은 상대적으로 둔감하게 반응하였다. 또한, 돈 및 우육의 n과 m 값 관계를 통해 기질 농도에 따른 저해 특성은 반경쟁적 저해 특성으로 판단된다.

• Archives of Pharmacal Research
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• 제19권4호
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• pp.258-260
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• 1996

Palmatine, an protoberberine isoquinoline alkaloid, has been found to inhibit dopamine biosynthesis by reducing tyrosine hydroxylase (TH) activity in PC12 cells (Lee and Kim, 1996). We have therefore investigated the effects of palmatine on bovine adrenal TH. Palmatine showed a mild inhibition on bovine adrenal TH (36.4% inhibition at concentration of $200\muM$). Bovine adrenal TH was inhibited competitively by palmatine with a substrate L-tyrosine. The Ki value was found to be 0.67 mM. This result suggests that the inhibition of TH activity by palmatine may be partially involved in the reduction of dopamine biosynthesis in PC12 cells.

• Bulletin of the Korean Chemical Society
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• 제22권11호
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• pp.1236-1242
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• 2001

(3S,1'S)-3-(1'-Carboxy-2'-phenyl)ethylamino-2-oxetanone (1a) and (3R,1'S)-3-(1'-carboxy-2'-phenyl)ethylamino-2-oxetanone (1b) were designed, synthesized, and evaluated as inhibitors for carboxypeptidase A, a prototypical zinc protease that removes the C-terminal amino acid having an aromatic side chain from oligopeptide substrate. It was concluded from the analysis of inhibition kinetics that while 1a inactivates CPA irreversibly, its diastereoisomer, 1b is a weak competitive inhibitor for CPA. A possible explanation for the observed difference in inhibition mode that is dependent on the inhibitor stereochemistry is offered.

• Biomolecules & Therapeutics
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• 제1권2호
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• pp.166-170
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• 1993

Hirudin is a potent inhibitor of thrombin, which was originally obtained from the medicinal leech (Hirudo medicinalis) Now it is being produced through the recombinant technology on a large scale. Recombinant hirudin has been assayed for the anticoagulant activity by the measurement of clotting time and the inhibition of thrombin actvity using a chromogenic substrate. The assay range of partial thromboplastin time and thrombin time is within $0.2{\sim}1.0 {\mu}g/mι.$ Thrombin time is more sensitive to the measurement of clot. Ex vivo study showed the level of hirudin in rat plasma was highest in 10 min and then it was eliminated slowly. The half-life of r-hirudin was 80~110 min depending on the assay methods. Intraveneous injection of russel viper venom was used for thrombus induction combined with vents cava ligation. Inhibition of venous thrombosis was observed with i.v. hirudin. It was dependent on the concentration of hirudin.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.179-182
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• 2001

Two intracellular enzymes, cellobiose phosphorylase (CBP) and cellodextrin phosphorylase (CDP) are involved in the phosphorolytic pathway in cellulose degradation. Those enzymes are considered to be useful in syntheses of oligosaccharides because the reactions are reversible. CBP from Cellvibrio gilvus and CDP from Clostridium thermocellum YM-4 were cloned and over-expressed in Escharichia coli. Both the enzyme reactions showed ordered bi bi mechanism. Acceptor specificity of CBP in the reverse reaction was determined. Several $\beta$-l,4-glucosyl disaccharides were synthesized by using the reaction. A new substrate inhibition pattern, competitive substrate inhibition, was also found in the reverse reaction of CBP Cellobiose was produced from sucrose at a high yield by a combined action of three enzymes including CBP