• 제목/요약/키워드: Enzyme-inhibition

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Enzyme Immunoassay for the Sulfamethazine Residues in Pork Tissue

  • Park, Jun-Hong;Lim, Yoon-Kyu
    • 한국식품위생안전성학회지
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    • 제11권4호
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    • pp.287-290
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    • 1996
  • To control the maximum residue level (MRL) for sulfamethazine (SMZ) residues in pork tissue, a microbial inhibition method is a regulatory screening assay method in Korea. Microwell plate-based competitive enzyme immunoassay (ELISA) kit is avalable for routine screening of SMZ residues in pork tissue. One ELISA kit is evaluated. Phosphate buffer extracts of samples fortified with SMZ at 0, 1, 5, and 10 ng/g were used in a recovery test of the kit. Market pork samples were assayed by the kit. Recovery of sulfamethazine was 104% at 10 ng/g. Intraassay variations and interassay variations for the kit were 7.70% and 5.76%, respectively. Concentration causing 50% inhibition of color development compared with blanks was 16.4ng. The violative pork samples with over MRL (0.1 $\mu\textrm{g}$/g) was 4 of 32 cases (12.5%) by used ELISA kit. This result indicates a possibility of the ELISA kit for screening test of SMZ residues in pork tissue, and still needs a comfirmatory assay for mandatory purposes.

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아세트알데히드로 유도된 급성독성에 대한 인삼부탄올 분획의 방어작용 (Preventive Effect of Ginseng Butanol Fraction against Acetaldehyde - Induced Acute Toxicity)

  • Keun Huh;Tae
    • Journal of Ginseng Research
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    • 제13권1호
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    • pp.5-7
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    • 1989
  • The objective of this study was to investigate the preventive effect of ginseng on acetaldehyde-induced acute toxicity in mice . Compared to the control group, treatment with acetaldehyde inhibited the hepatic cytosolic xanthine oxidise activity with increase in dose. The inhibition of enzyme activity was not changed after dialysis. Pretreatment with ginseng butanol fraction prevented the inhibition of enzyme activity by acetaldehyde. In conjunction with the our previous results (Yakhak Hoeji, 29, 18 (1985)), these results suggest that the most likely mechanism for the observed preventive effects of ginseng against the acetaldehyde-induced acute toxicity may be the decrease hepatic acetaldehyde level.

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생리활성과 분자구조의 상관관계에 관한 연구 (The Studies in Relationship between Molecular Structure and Biological Activities)

  • 김의락;민경섭;김종토;정봉진
    • 대한화학회지
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    • 제37권1호
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    • pp.68-75
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    • 1993
  • 분자구조를 기술하는 molecular connectivity index, Wiener index 및 ad hoc descriptor와 알코올, 에스테르, 케톤 화합물들이 생체내에서 나타내는 enzyme inhibitory potency, lipoxygenase inhibition, tadpole narcosis potency, 증기독성(vapor toxicity), 증발열(heat of vaporization)과 같은 성질들 사이의 상관관계를 조사하였다. 생물학적 활성의 종류에 따라 molecular descriptor 사이의 우열은 있으나, 대체로 좋은 상관관계식을 얻었다.

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Studies on the development of enzyme linked immuno-sorbent assay (ELISA) for hepatitis B surface antigen (HBsAg) by monoclonal antibodies of different affinity constants

  • Kim, Gye-Won;Hong, Sung-Youl;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • 제10권1호
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    • pp.18-24
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    • 1987
  • Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

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Alteration of hepatic anti-oxidant systems by 4-nonylphenol, a metabolite of alkylphenol polyethoxylate detergents, in Far Eastern catfish Silurus asotus

  • Park, Kwan Ha
    • Environmental Analysis Health and Toxicology
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    • 제30권
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    • pp.6.1-6.7
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    • 2015
  • Objectives This study aimed to estimate the effects of 4-nonylphenol (NP), a ubiquitously present surfactant in aquatic environments, on the anti-oxidant systems of the liver in the Far Eastern catfish Silurus asotus. Methods Changes in biochemical parameters involved in glutathione (GSH)-related and other anti-oxidant systems were analyzed following 4 weeks of 4-NP administration (0.1 and 1.0 mg/kg diet) via a formulated diet to catfish. Results 4-NP exposure induced an elevation in hepatic lipid peroxide levels and an accompanying decrease in reduced state GSH after 2 weeks, suggesting pro-oxidant effects of the chemical in catfish. This oxidative stress was associated with an inhibition of the GSH-utilizing enzyme glutathione peroxidase at the same time point. This inhibition was restored after 4 weeks. The activities of other anti-oxidant enzymes, i.e., glutathione reductase, superoxide dismutase and catalase were increased after 4 weeks. These enzyme increases occurred more strongly at the higher 4-NP concentration (1.0 mg/kg diet). Conclusions 4-NP given to catfish at 0.1 to 1.0 mg/kg diet, concentrations relevant to environmental levels, depletes the endogenous anti-oxidant molecule GSH and temporarily inhibits GSH-related anti-oxidant enzymes. Such declines in anti-oxidant capacity and elevated oxidative stress seem to be compensated eventually by subsequent activation of various anti-oxidant enzyme systems.

Effect of phlorotannins isolated from Ecklonia cava on angiotensin I-converting enzyme (ACE) inhibitory activity

  • Wijesinghe, W.A.J.P.;Ko, Seok-Chun;Jeon, You-Jin
    • Nutrition Research and Practice
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    • 제5권2호
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    • pp.93-100
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    • 2011
  • Inhibition of angiotensin I-converting enzyme (ACE) activity is the most common mechanism underlying the lowering of blood pressure. In the present study, five organic extracts of a marine brown seaweed Ecklonia cava were prepared by using ethanol, ethyl acetate, chloroform, hexane, and diethyl ether as solvents, which were then tested for their potential ACE inhibitory activities. Ethanol extract showed the strongest ACE inhibitory activity with an $IC_{50}$ value of 0.96 mg/ml. Five kinds of phlorotannins, phloroglucinol, triphlorethol-A, eckol, dieckol, and eckstolonol, were isolated from ethanol extract of E. cava, which exhibited potential ACE inhibition. Dieckol was the most potent ACE inhibitor and was found to be a non-competitive inhibitor against ACE according to Lineweaver-Burk plots. Dieckol had an inducible effect on the production of NO in EAhy926 cells without having cytotoxic effect. The results of this study indicate that E. cava could be a potential source of phlorotalnnins with ACE inhibitory activity for utilization in production of functional foods.

The effective model of the human Acetyl-CoA Carboxylase inhibition by aromatic-structure inhibitors

  • Minh, Nguyen Truong Cong;Thanh, Bui Tho;Truong, Le Xuan;Suong, Nguyen Thi Bang;Thao, Le Thi Xuan
    • 전기전자학회논문지
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    • 제21권3호
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    • pp.309-319
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    • 2017
  • The research investigates the inhibition of fatty acid biosynthesis of the human Acetyl-CoA Carboxylase enzyme by the aromatic-structure inhibitors (also known as ligands) containing variables of substituents, contributing an important role in the treatment of fatty-acid metabolic syndrome expressed by the group of cardiovascular risk factors increasing the incidence of coronary heart disease and type-2 diabetes. The effective interoperability between ligand and enzyme is characterized by a 50% concentration of enzyme inhibitor ($IC_{50}$) which was determined by experiment, and the factor of geometry structure of the ligands which are modeled by quantum mechanical methods using HyperChem 8.0.10 and Gaussian 09W softwares, combining with the calculation of quantum chemical and chemico-physical structural parameters using HyperChem 8.0.10 and Padel Descriptor 2.21 softwares. The result data are processed with the combination of classical statistical methods and modern bioinformatics methods using the statistical softwares of Department of Pharmaceutical Technology - Jadavpur University - India and R v3.3.1 software in order to accomplish a model of the quantitative structure - activity relationship between aromatic-structure ligands inhibiting fatty acid biosynthesis of the human Acetyl-CoA Carboxylase.

Lysosomal Enzyme Inhibiting Activity of Alkaloidal Fraction from Tylophora indica Leaves in Arthritic rats

  • Arora, Sandeep;Singh, Hemant Kumar
    • Natural Product Sciences
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    • 제13권4호
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    • pp.289-294
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    • 2007
  • Ethanolic extract (50% v/v) and alkaloid fraction of Tylophora indica leaves were examined for lysosomal enzyme inhibitory activity in adjuvant-induced arthritic rats. The alkaloid fraction showed statistically significant inhibition of arthritic lesions (p < 0.05) from day 18, (p < 0.025) from day 20 and (p < 0.001) from day 21 onwards in the adjuvant-induced arthritis, which was comparable to the response of standard drug Indomethacin. The ethanolic extract was less significant than the alkaloidal fraction in inhibition of arthritis. Alkaloid fraction showed significant (p < 0.001) inhibitory effect on the lysosomal enzyme activities in adjuvantinduced arthritic rats. It also significantly prevented decrease in collagen levels and synovial damage observed during arthritis and also inhibited increase in urinary excretion levels of collagen degradation products like hydroxyproline, hexosamine, hexuronic acid, etc. Both ethanolic extract as well as the alkaloid fraction, however, did not show any significant activity in normal nonarthritic rats. The ethanolic extract and the alkaloid fraction may thus be able to inhibit the progress of inflammation and inhibit the destructive activity of lysosomal enzymes on structural macromolecules like collagen etc. in the synovial capsule in joints during arthritic states. They may thus prevent synovial damage observed during arthritis.

Structure Characterization and Antihypertensive Effect of an Antioxidant Peptide Purified from Alcalase Hydrolysate of Velvet Antler

  • Seung Tae Im;Seung-Hong Lee
    • 한국축산식품학회지
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    • 제43권1호
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    • pp.184-194
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    • 2023
  • Recently, interest in food-derived bioactive peptides as promising ingredients for the prevention and improvement of hypertension is increasing. The purpose of this study was to determine the structure and antihypertensive effect of an antioxidant peptide purified from velvet antler in a previous study and evaluate its potential as a various bioactive peptide. Molecular weight (MW) and amino acid sequences of the purified peptide were determined by quadrupole time-of-flight electrospray ionization mass spectroscopy. The angiotensin I-converting enzyme (ACE) inhibition activity of the purified peptide was assessed by enzyme reaction methods and in silico molecular docking analysis to determine the interaction between the purified peptide and ACE. Also, antihypertensive effect of the purified peptide in spontaneously hypertensive rats (SHRs) was investigated. The purified antioxidant peptide was identified to be a pentapeptide Asp-Asn-Arg-Tyr-Tyr with a MW of 730.31 Da. This pentapeptide showed potent inhibition activity against ACE (IC50 value, 3.72 μM). Molecular docking studies revealed a good and stable binding affinity between purified peptide and ACE and indicated that the purified peptide could interact with HOH2570, ARG522, ARG124, GLU143, HIS387, TRP357, and GLU403 residues of ACE. Furthermore, oral administration of the pentapeptide significantly reduced blood pressure in SHRs. The pentapeptide derived from enzymatic hydrolysate of velvet antler is an excellent ACE inhibitor. It might be effectively applied as an animal-based functional food ingredient.

Kinetic Study on the Enzymatic Production of D-Alanine from D-Aspartic Acid

  • Lee, Jae-Heung;Sung, Moon-Hee;Jeon, Yeong-Joong
    • Journal of Microbiology
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    • 제40권1호
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    • pp.33-37
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    • 2002
  • An enzymatic reaction for the production of D-alanine from D-aspartic acid and pyruvate as substrates by a thermostable D-amino acid aminotransferase (D-AAT) was investigated at various conditions In the temperature range of 40-70$\^{C}$ and pH range of 6.0-9.5. The D-AAT was produced with recombinant E. coli BL21, which hosted the chimeric plasmid pTLK2 harboring the D-AAT from the novel thermophilic Bacillus sp. LK-2. The enzyme reaction was shown to follow the Ping Pong Bi Bi mechanism. The K$\_$m/ values for D-aspartic acid and pyruvate were 4.38 mar and 0.72 mM, respectively. It was observed that competitive inhibition by D-alanine, the product of this reaction, was evident with the inhibition constant K$\_$i/ value of 0.1 mM. A unique feature of this reaction scheme is that the decorboxylation of oxaloacetic acid, one of the products, spontaneously produces pyruvate. Therefore, only a catalytic amount of pyruvate is necessary for the enzyme conversion reaction to proceed. A typical time-course kinetic study skewed that D-alanine up to 88 mM could be produced from 100 mM of D-aspartic acid with a molar yield of 1.0.