• Journal of Life Science
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• 제11권2호
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• pp.108-110
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• 2001

The E. coli expression system to overproduce a harmful protein, carboxypeptidase Taq was developed. Since expression plasmid pCK305N containing the colicin promoter already has the initiation codon on the restriction site, the initiation codon of the CPase Taq gene was removed. Expression plasmid pCP4-col includes the entire CPase Taq gene, which is directed by the colicin promoter. E. coli cells harboring pCP-col produced a high amount of the enzyme when they were cultured in the present of mitomycin C (0.4 ${\mu}g$/ml). An amount of purified enzyme produced by pCP4-col directed by the colicin promoter was 10.5 mg. This result indicated that the novel E. coli expression system controlled by the colicin promoter could produce almost twice amounts of CPase Taq than the conventional system controlled by the tart promoter.

• The Korean Journal of Physiology and Pharmacology
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• 제18권3호
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• pp.269-278
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• 2014

Various antiepileptic drugs (AEDs) especially enzyme-inducing AEDs might be associated with increased vascular risk, through impairment of the endogenous antioxidative ability which may trigger oxygen-dependent tissue injury. Lamotrigine (LTG) a non-enzyme-inducing AED has scarce information regarding its effects on oxidative stress. The present study aimed to study the possible modulation of vascular risk factors of epileptogenesis by LTG, in a rat model of kindling seizure induced by pentylenetetrazole (PTZ). Four groups of male Wister rats were used; vehicle control group, PTZ group (alternate day PTZ, 30 mg/kg, i.p), LTG/PTZ group (LTG 20 mg/kg/day p.o and alternate day PTZ) and LTG group. The study period was 5 weeks. Lipoproteins and total homocysteine (tHcy), malondialdehyde (MDA) and reduced glutathione (GSH) were measured. Aortic endothelial function study and histopathological examination of the rats' brains, aortas and coronaries were conducted. Serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C), tHcy, MDA, GSH levels were significantly higher in epileptic rats than normal controls rats. A decrease in HDL-cholesterol with high atherosclerotic index was also demonstrated. The administration of LTG improved the PTZ-kindled seizures. It produced a significant decrease in TC, TG and LDL-cholesterol, MDA, aortic GSH and increase in HDL-cholesterol with no significant effect on serum GSH and tHcy levels. LTG improved endothelium-dependent relaxation, decreased hippocampal neurodegenerative changes and atherosclerotic changes of aortas and coronaries. LTG decreased seizures severity, hippocampal damage and improved vascular risk markers in this rat model of kindling seizures.

• Biomolecules & Therapeutics
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• 제8권4호
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• pp.299-304
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• 2000

The modulation of cytochrome P450(P450) activities and glutathione S-transferase (GST) was investigated after i.p. administration of glucose-diethyldithiocarbamate (Glu-DDTC) to rats. P450 1 A2 and 2El activities were inhibited by 60% 4 hr after the administration of 200 mg Glu-DDTC/kg and those activities were recovered to original levels 24 hr after dosing. In contrast, GST activities were enhanced up to 24 hr after dosing. These results seem to be due to the bifunctional activity of Glu-DDTC. Glu-DDTC acts as an inhibitor of P450 enzymes as well as inducer of GST enzyme. Glu-DDTC inhibited PNP hydroxylation (P450 2El) and ethoxycoumarin O-deethylation (P450 1A2) in a dose-dependent manner up to 200 mg/kg wherease it did not affect testosterone 6$\beta$-hydroxylation (P450 3A) and pentoxyresorufin O-dealkylation (P450 2B) activities. Induction of GST activity toward 1-chloro-2,4-dinitrobenzene (CDNB) and 1,2-dichloro-4-nitrobenzenen (DCNB) was dependent on the dose of Glu-DDTC and no species difference in the GST induction was seen between rat and mouse. Amoung GST subunits, Ya, Yb1 and partially Yb2 were induced by Glu-DDTC as conjugated by western blotting. The levels Yp, Yk and Yc subunits were not affected by Glu-DDTC treatment. Therefore the enhanced activity of GST toward CDNB and DCNB might be due to the induction of Ya, Ybl and partially Yb2 subunits. In conclusion, Glu-DDTC selectively inhibited P45O 1A2 and P450 2El activities whereas it enhanced Ya, Ybl subunits and partially Yb2 subunits of GST and the antimutagenic activity of this compound might be attributed from the modulation of these enzyme activities in animals.

• Applied Biological Chemistry
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• 제20권1호
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• pp.33-42
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• 1977

콩나물에 많이 존재하는 Asn의 생합성(生合成)에 관하여 연구하고자 콩나물 생육기간중(生育其間中) 수분함량(水分含量) 및 Asn 함량을 조사한 결과 10일째 수분(水分)은 $85{\sim}90%$에 달했으며 Asn은 6일째부터 급격히 증가하여 10일째는 건물량(乾物量)의 22.7%에 달하였다. 콩나물 100 g에서 Asn을 추출(抽出)한 결과 2.29g을 얻었고 재결정하여 1.62 g을 얻었다. 콩나물 생육시(生育時) $NH_4Cl,\;(NH_4)_2SO_4$ 및 urea를 살포하여 질소화합물에 대한 영향을 고찰하였던 바 $NH_4Cl$ 및 $(NH_4)_2SO_4$구(區)는 Asn이 control보다 약간 증가하였으나 urea구(區)는 10일째 29.5%로 증가하였다. 콩나물에서 asparagine synthetase를 $(NH_4)_2SO_4$ fractionion 및 Sephadex G-150에 의해 정제하여 8.6배 정제하였다. 이 효소는 매우 불안정(不安定)하였으나 glycerol, mercaptoethanol 및 기질(基質)인 ATP, $Mg^{++}$이온에 의해 보호를 받았으며 이 산소(酸素)에 의해 최고(最高)의 Asn이 생성(生成)되는 ATP, Gln(또는 ${NH_4}^+$)의 농도는 표준반응 용액에서 각각의 구성농도를 변화시키면서 측정하였을 때 ATP 10 mM, ${NH_4}^+$는 50 mM, Gln은 10 mM이었다. $Mg^{++}$이온 대신 $Mn^{++}$, $Zn^{++}$, 및 $Fe^{++}$로 대치할수 없었으며 Asp에 대한 Km값은 3.1 mM이었고 효소작용의 최적(最適) pH는 7.5이었다. Amide donor로서 $NH_2OH$로 대치한 결과 ${\beta}-aspartyl$ hydroxamate가 생성(生成)되어, 효소의 열불안성(熱不安性), amide donor로서 Gln 이용 및 SH기(基)에 의한 효소안정성을 비추어 보아, 같은 두과식물(荳科植物)인 완두와 비슷하게 asparagine synthetase는 ${\beta}-aspartyl$ adenylate의 중간반응(中間反應)을 거치는 기작(機作)에 의해 Asn을 생합성(生合成)하는 것을 알았다.

• Bulletin of the Korean Chemical Society
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• 제25권5호
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• pp.721-725
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• 2004

Acetohydroxy acid synthase (AHAS) is one of several enzymes that require thiamine diphosphate and a divalent cation as essential cofactors. The active site contains several conserved ionizable groups and all of these appear to be important as judged by the fact that mutation diminishes or abolishes catalytic activity. Recently, we have shown [Yoon, M.-Y., Hwang, J.-H., Choi, M.-K., Baek, D.-K., Kim, J., Kim, Y.-T., Choi, J.-D. FEBS Letters 555 (2003), 185-191] that the activity is pH-dependent due to changes in $V_{max}$ and V/$K_m$. Data were consistent with a mechanism in which substrate was selectively catalyzed by the enzyme with an unprotonated base having a pK 6.48, and a protonated group having a pK of 8.25 for catalysis. Here, we have in detail studied the pH dependence of the kinetic parameters of the cofactors (ThDP, FAD, $Mg^{2+}$) in order to obtain information about the chemical mechanism in the active site. The $V_{max}$ of kinetic parameters for all cofactors was pH-dependent on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 9.5, 9.3 and 10.1, respectively. The V/$K_m$ of kinetic parameters for all cofactors was pH-dependent on the acidic and on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 6.2-6.4 on the acidic side and 9.0-9.1 on the basic side. The well-conserved histidine mutant (H392) did not affect the pH-dependence of the kinetic parameters. The data are discussed in terms of the acid-base chemical mechanism.

• 한국식품영양학회지
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• 제20권2호
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• pp.114-119
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• 2007

Our research objective was to examine the in vitro biological activity of deer antler(Nogyong in Korean) extract, including the antioxidative, nitrite scavenging, and tyrosinase inhibitory effects, as well as the antithrombotic, and angiotensin I converting enzyme(ACE) inhibitory activities. The carbohydrate, protein, fat, and mineral contents of the deer antler were 7.6%, 65.3%, 3.2% and 23.9%, respectively. The electron donating ability(EDA) by the reduction of 2,2'-diphenyl-1-picrylhydrazyl(DPPH) was 67.1%, and the inhibition rate of lipid peroxidation by the thiocyanate method using linoleic acid was 92.1% in 100 mg/ml of extract. The nitrite scavenging effects were pH dependent, and were highest at pH 1.2 and lowest at pH 6.0. The sample inhibition rate against tyrosinase was above 64.0%. The platelet aggregation induced by ADP(adenosine-5'diphosphate) was inhibited up to 51.7%, and the inhibitory effect was dependent on the sample concentration. Lastly, the inhibition rate of ACE was 47.5% in 100 mg/ml of deer antler extract.

• 동의신경정신과학회지
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• 제19권3호
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• pp.129-142
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• 2008

Objective: The purpose of this study was to evaluate the antioxidative effects of the extract of Scolopendra subspinipes which has been used mainly for detoxication in the oriental medicine and reported to have sedative action, antiinflammatory effect, antihypertensive property and immunity enhancing activity. Method: Inhibitory activities on oxygen radical generating enzymes (aldehyde oxidase and xanthine oxidase) and increasing activities on oxygen radical scavenging enzymes (superoxide dismutase, glutathione peroxidase, glutathione-S-transferase) were investigated. Furthermore, the content of glutathione in the mouse brain, DPPH radical scavenging activity and also anti-lipid peroxidative effects in vivo and in vitro were estimated. Results: The extract showed weak inhibitory effects on the activities of aldehyde oxidase and xanthine oxidase which are oxygen radical generating enzymes. The extract inhibited lipid peroxidation with 26.1% against control group at 500 mg/kg in vivo and with 11.2% against control group at 10 mg/kg in vitro in a dose-dependent manner, which means this drug may protect radical-induced cell damages. The extract showed dose-dependently the scavenging effect on DPPH radical with 24.8% activity at 10 mg/ml in vitro. The extract enhanced the activities of superoxide dismutase, glutathione peroxidase and glutathione-S-transferase, which are oxygen radical scavenging enzymes, with 28.9%, 22.3% and 23.1%, respectively at 500mg/kg in vivo. Finally, this extract strongly increased the glutathione content in the mouse barin. Conclusion: Above results indicated that Scolopendra subspinipes can be useful for the protection or treatment of some diseases caused by reactive oxygen species.

• 생약학회지
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• 제53권4호
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• pp.213-225
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• 2022

Schisandra chinensis is widely known to regulate fatigue recovery, lowering blood pressure, and contains several bioactive compounds such as schizadran. In this study, we obtained basic data for the development of health food by measuring the schizandrin content, antioxidant activity, anti-inflammatory, antibacterial activity, and enzyme activity of the hot water extract (SCW) and 40% ethanol extract (SCE) of S. chinensis grown in Sunchang-gun, Korea. Schizandrin content was measured as 7.87 ± 0.01 mg/g for SCW and 10.38 ± 0.05 mg/g for SCE. SCE had higher DPPH and ABTS radical scavenging activity than SCW, and the total polyphenol and flavonoid contents were also higher. Both SCW and SCE (500 ㎍/mL) exhibited more than 55% protective effects against oxidative stress in HepG2 cell lines. Anti-inflammatory efficacy was assessed using RAW 264.7 and Caco-2 cells and both SCW and SCE do not have cytotoxic effects. Anti-inflammatory results showed concentration-dependent NO activity and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) inhibition. The antibacterial activity increased with increasing dose concentrations of SCW and SCE, and the MIC was 25 mg/mL for L. monocytogenes, S. typhimurium, and 75 mg/mL for H. pylori. In addition, amylase and protease enzyme activity was observed in both SCW and SCE.

• Journal of Ecology and Environment
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• 제34권3호
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• pp.315-322
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• 2011

We investigated the effect of Pb uptake by Pinus densiflora and the Pb fraction in forest soil. We also investigated the change in soil physicochemical characteristics, microbial activity, and root exudates of Pinus densiflora in Pb-contaminated soils. Three-year-old pine seedlings were exposed to 500 mg/kg Pb for 12 months. The metal fractions were measured using sequential extraction procedures. Additionally, factors that affect solubility (three soil enzyme activities and amino acids of root exudate compounds) were also determined. The results showed that Pb contamination significantly decreased enzyme activities due to soil characteristics. In addition, organic matter, nitrate content, and Pb concentration were time dependent. The results indicate that changes in the Pb fraction affected Pb uptake by pine trees due to an increase in the exchangeable Pb fraction. The concentrations of organic acids were higher in Pb-spiked soil than those in control soil. Higher rhizosphere concentrations of oxalic acid resulted in increased Pb uptake from the soil. These results suggest that pine trees can change soil properties using root exudates due to differences in the metal fraction.

• 한국식품저장유통학회지
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• 제30권6호
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• pp.905-917
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• 2023

The optimal fermentation conditions for producing fermented black soybeans rich in sulfur-containing amino acids were investigated. Response surface methodology monitored fermentation conditions. A central composite design examined the effect of independent variables: enzyme concentration (X₁) and fermentation time (X₂) on yield and methionine content. Both factors significantly influenced these dependent variables. Enzyme concentration more profoundly affected amino nitrogen content than fermentation time. β-Glucan content and cystine level were primarily affected by fermentation time. We elicited each variable's regression formula and identified optimal fermentation conditions for functional compounds. The predicted optimum conditions were an enzyme concentration of 0.28-0.32% and a fermentation time of 58.0-62.0 min. Under these optimal conditions, each black soybean variety's sulfur amino acid content ranged from 818.62 to 922.62 mg/100 g, demonstrating significant variety differences.