• Korean Journal of Microbiology
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• 제31권3호
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• pp.224-229
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• 1993

GABA transminase (4-aminobutyrate aminotransferase), which catalyzes the breakdown of the major inhibitory neurotransmitter, GABA, in mammalian brain, was inactivated by preincubation with the mycotoxin patulin. The time course of the reaction was significantly affected by the substrate .alpha.-ketoglutarate, which aforded complete protection against the loss of catalytic activity. The recovery from the inhibition of patulin by the addition of dithiothreitol (DTT) supports that patulin reacts with the sulfhydryl residue in the catalytic domain of the enzyme. The reconstitution of the reduced enzyme and apoenzyme with pyridoxal-5-P(PLP) was inhibited by another mycotoxin, penicilic acid. This mycotoxin may interact with lysyl residue of the enzyme. Therefore, it is postulated that the critical sulfhydryl and lysyl residues in the catalytic domain of the enzyme react with mycotoxin patulin and penicillic acid, respectively.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 한국지하수토양환경학회 1998년도 공동 심포지엄 및 추계학술발표회
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• pp.174-177
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• 1998

The effects of surfactants affecting polycyclic Aromatic Hydrocarbon(PAHs) solubility and biodegradation in soil slurry were investigated. The critical micelle concentration(CMC) values of surfactants used in this study were 12.7mg/L(Brij 30), 13.4mg/L(Tween 80), 13.6mg/t(Triton X-100). The solubility of PAH increased as the Hydrophile-Lipophile Balance(HLB) value of surfactant decrease. At surfactant biodegradation and toxicity experiement using respirometer, Brij 30 did not show any toxic effect and substrate inhibition upon the level of 1.5g/L. Also, biodegradation of Brij 30 gave no reduction on the phenanthrene biodegradation rate. When the desorption rate of phenanthrene between sand and clay is compared, lower percentage of phenanthrene was desorbed at clay because of the larger surface aera and higher organic content of clay. At the biodegradation experiments of phenanthrene in soil slurry phase, more than 90% of initial phenanthrene adsorbed onto both sand and clay were biodegraded by phenanthrene- acclimated cultures.

• Journal of the Korean Society of Tobacco Science
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• 제21권1호
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• pp.49-56
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• 1999

Classification of esterase isozymes of the apterous green peach aphids (Myzus persicae Sulzer) collected in tobacco fields were investigated by the native polyacrylamide gel electrophoresis (PAGE). A total of twelve esterase bands were identified in adult apterous aphid, and the difference of enzyme band activity in the clones was observed at the first and second bands group. Esterases of green peach aphids reacted with specific substrate were more stained $\alpha$-naphthyl acetate than $\alpha$-naphthyl propionate, and $\alpha$-naphthyl acetate more than $\beta$-naphthyl acetate. Twelve esterases on the basis of inhibition by the three types of inhibitors (organophosphates: 2.5$\times$10$^{-3}$ M paraoxon, 4$\times$10$^{-3}$ M DFP; eserine sulfate : 2$\times$10$^{-3}$ M eserin; sulfhydryl reagents: 2$\times$10$^{-3}$ M p-HMB) were classified into three class, namely, cholinesterase (ChE) I, II, carboxylesterase (CE) and arylesterase (ArE), and these classes contained 3, 4, 3 and 2 isozymes, respectively.

• Biomedical Science Letters
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• 제12권3호
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• pp.225-231
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• 2006

Fibrinolytic enzyme was purified from the fruiting bodies of Lepista nuda, using DEAE-Cellulose chromatography, Phenyl Sepharose chromatography, and Mono-S column chromatography. The substance has a molecular weight of 30006.62 Da as measured by MALD-TOF mass spectrometry. The N-terminal amino acid sequence of the enzyme was Tyr-Pro-Ser-Pro-Ser-His-Gln-Thr-Ala-Val-Asn-Ala-Ile-Ile-X. The activity of the enzyme was inhibited by PMSF, indicating that the enzyme is a serine protease. No inhibition was found with E-64, pepstatin, and EDTA. It has broad substrate specificity for synthetic peptides. The enzyme was stable up to $30^{\circ}C$. The enzyme hydrolyzes both Aa and y chains of human fibrinogen but did not show any reactivity for $B{\beta}$ chain of human fibrinogen.

• Archives of Pharmacal Research
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• 제21권2호
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• pp.207-211
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• 1998

Three prolyl endopeptidase inhibitors were isolated and identified as luteolin, quercetin and ${\beta}$-sitosterol-3-O-${\beta}$-D-glucopyranoside with $IC_{50}$ of 0.17, 0.19 and 27.5 ppm, respectively. The inhibition of two flavonoids were non-competitive with substrate. Twenty authentic flavonoids were tested in order to investigate structure-activity relationship. No significant relationship was found in them, however, catechol moiety of B-ring and 7-OH group in flavonoid skeleton were seemed to be responsible for the stronger activity.

• Archives of Pharmacal Research
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• 제17권5호
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• pp.354-359
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• 1994

The activity of SD-framesylcysteine O-methyltransferase was assayed by incubating the enzyrne with a synthetic in vitro substrate, [N-acetyl-S-trans, trns-famesyl-L-cysteine (AFC)], together with S-adenosyl-L-[emthyl-$_{14}$C)ester(AFCME)], was then analyzed either directly on HPLC or by converting the AFC[$methyl^{14}C$]ME to [$methyl^{14}C$] aclcohol by basehydrolysis. Employing these two analytical methods, it was established that a peptide purifed from rat liver cytosol fraction [Int. J. Biochem., 25, 1157 919930] strongly inhibited the above enzyme activity with $IC_{50}\; of\; 7.1\times 10^{-8}$ M. Also, the S-famesylcysteine O-methyltransferase from several human colon cancer cells was equally inhibited by the peptide.

• Toxicological Research
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• 제22권1호
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• pp.39-45
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• 2006

As the antioxidant and free radical scavenger, glutathione (GSH) participates in the preservation of cellular redox status and defense against reactive oxygen species and xenobiotics. Glutamate-cysteine ligase (GCL; also known as ${\gamma}$-glutamylcysteine synthetase, EC 6.3.2.2) is the rate limiting enzyme in GSH synthesis. In the present study, the accurate method for determination of GCL activity in crude liver extracts was developed by measuring both ${\gamma}$-glutamylcysteine and GSH from cysteine in the presence of glutamate, glycine and an ATP-generating system. We added glycine to promote the conversion of ${\gamma}$-glutamylcysteine to GSH, and to minimize the possibility of ${\gamma}$-glutamylcysteine metabolism to cysteine and oxoproline by ${\gamma}$-glutamylcyclotransferase. We established optimal conditions and substrate concentrations for the enzyme assay, and verified that inhibition of GCL by GSH did not interfere with this assay. Therefore, this assay of hepatic GCL under optimal conditions could provide a more accurate measurement of this enzyme activity in the crude liver extracts.

• BMB Reports
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• 제37권5호
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• pp.515-521
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• 2004

The tricarboxylate (or citrate) carrier was purified from eel liver mitochondria and functionally reconstituted into liposomes. Incubation of the proteoliposomes with various sulfhydryl reagents led to inhibition of the reconstituted citrate transport activity. Preincubation of the proteoliposomes with reversible SH reagents, such as mercurials and methanethiosulfonates, protected the eel liver tricarboxylate carrier against inactivation by the irreversible reagent N-(1-pyrenyl)maleimide (PM). Citrate and L-malate, two substrates of the tricarboxylate carrier, protected the protein against inactivation by sulfhydryl reagents and decreased the fluorescent PM bound to the purified protein. These results suggest that the eel liver tricarboxylate carrier requires a single population of free cysteine(s) in order to manifest catalytic activity. The reactive cysteine(s) is most probably located at or near the substrate binding site of the carrier protein.

• Journal of the Korean Institute of Intelligent Systems
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• 제3권3호
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• pp.3-18
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• 1993

본 논문에서는 박테리아에서 생성되는 생체 계면활성제인 emulsan의 생산을 위한 유가식 배양에서 에칸을 농도의 제어에 퍼지기법을 적용하였다. 기절저해가 있는 유가식 배양에서 emulsan의 생산을 향상시키기 위해 최대 비성장속도를 갖는 최적 기질농도가 유지되도록 기질인 에탄올의 공급 속도가 조절되어 졌다. 생물반응기에서 Acunetobacter calcoaceticus RAG-1 박테리아를 회분식과 유가식으로 배양 실험하여 최적 에탄올 농도를 구하고, kinetic 모델을 제시하였다. 배양실험의 결과와 지식을 바탕으로 퍼지 규칙을 구성하였다. 퍼지 제어기에서 제어 입력변수는 기질농도의 최적치와 운전치의 오차와 오차의 변화로서 구성되고, 제어 출력변수는 기질 공급 속도의 변화량으로 구성되었다. 멤버쉽 함수를 입력변수의 퍼지 집합화 과정을 통하여 구하였고, 최소-최대법과 무게 중심법을 이용하여 출력 제어값을 구하였다. 유가식 배양의 전산모사와 실험 결과에서 퍼지제어 기법은 최적 기질 농도를 정확히 제어하였으며, emulsan 생산은 향상되었다.

• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.85-89
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• 1991

The aim of this study is to elucidate characteristics of ${\beta}-lactamase$ inhibitor produced by Streptomyces sp. SMF-19 isolated from soil was found to produce proteinaceous extracellular ${\beta}-lactamase$ inhibitor. The ${\beta}-lactamase$ inhibitor was purified through ammonium sulfate fractionation, gel filtration, anion exchange chromatography and fast performace liquid chromatography. The molecular weight of the ${\beta}-lactamase$ inhibitor was estimated to be about 48,000 by SDS-PAGE. The mode of inhibition against penicillin G as a substrate was uncompetitive. The ${\beta}-lactamase$ inhibitor was stable in wide pH range but unstable at high temperature above $50^{\circ}C$.