• Title/Summary/Keyword: isozymes

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Thermal Inactivation of Myrosinase from White Mustard Seeds

  • Ko, Young Hwan;Lee, Ran
    • The Korean Journal of Food And Nutrition
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    • v.34 no.1
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    • pp.26-35
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    • 2021
  • Myrosinases (thioglucosidases) catalyze the hydrolysis of a class of compounds called glucosinolates, of which the aglycones show various biological functions. It is often necessary to minimize the loss of myrosinase activity during thermal processing of cruciferous vegetables. Myrosinase was isolated from a popular spice, white mustard (Sinapis alba), and its thermal inactivation kinetics was investigated. The enzyme was extracted from white mustard seeds and purified by a sequential processes of ammonium sulfate fractionation, Concanavalin A-Sepharose column chromatography, and gel permeation chromatography. At least three isozymes were revealed by Concanavalin A-Sepharose column chromatography. The purity of the major myrosinase was examined by native polyacrylamide gel electrophoresis and on-gel activity staining with methyl red. The molecular weight of the major enzyme was estimated to be 171 kDa. When the consecutive step model was used for the thermal inactivation of the major myrosinase, its inactivation energy was 44.388 kJ/mol for the early stage of destruction and 32.019 kJ/mol for the late stage of destruction. When the distinct two enzymes model was used, the inactivation energy was 77.772 kJ/mol for the labile enzyme and 95.145 kJ/mol for the stable enzyme. The thermal inactivation energies lie within energy range causing nutrient destruction on heating.

CJ-11668, a new selective and potent cox-2 inhibitor, has long-acting pharmacokinetic profiles

  • Park, Hyun-Jung;Kang, Hye-Jung;Chung, Young-Mee;Chun, Hyung-Ok;Hong, Kwang-Hee;Kim, Il-Hwan;Kim, Taek-Rho;Noh, Hyun-Jung;Kim, Deog-Yeor
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.88.1-88.1
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    • 2003
  • CJ-11668 is a new potent and selective COX-2 inhibitor (IC$\sub$50/ COX-2 65nM; COX-l/COX-2 ratio 770). The pharmacokinetic profile of CJ-11668 (20 mg/kg, p.o.) in the rat was characterized by high bioavailability (90%) and long plasma half-life (11.7 hr) with low clearance (0.4 L/hr/kg). In the dog, the PK profiles (2 mg/kg, p.o.) also showed long plasma half-life (l7.9hr) with low clearance (0.5 L/hr/kg), and the bioavalability of 60%. The inhibition of CJ-11668 infive different cytochrome P450 isozymes (1A2, 2C9, 2C19, 2D6 and 3A4) was determined in vitro and had observed no significant effect. (omitted)

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CKD-581 Downregulates Wnt/β-Catenin Pathway by DACT3 Induction in Hematologic Malignancy

  • Kim, Soo Jin;Kim, Suntae;Choi, Yong June;Kim, U Ji;Kang, Keon Wook
    • Biomolecules & Therapeutics
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    • v.30 no.5
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    • pp.435-446
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    • 2022
  • The present study evaluated the anti-cancer activity of histone deacetylase (HDAC)-inhibiting CKD-581 in multiple myeloma (MM) and its pharmacological mechanisms. CKD-581 potently inhibited a broad spectrum of HDAC isozymes. It concentration-dependently inhibited proliferation of hematologic cancer cells including MM (MM.1S and RPMI8226) and T cell lymphoma (HH and MJ). It increased the expression of the dishevelled binding antagonist of β-catenin 3 (DACT3) in T cell lymphoma and MM cells, and decreased the expression of c-Myc and β-catenin in MM cells. Additionally, it enhanced phosphorylated p53, p21, cleaved caspase-3 and the subG1 population, and reversely, downregulated cyclin D1, CDK4 and the anti-apoptotic BCL-2 family. Finally, administration of CKD-581 exerted a significant anti-cancer activity in MM.1S-implanted xenografts. Overall, CKD-581 shows anticancer activity via inhibition of the Wnt/β-catenin signaling pathway in hematologic malignancies. This finding is evidence of the therapeutic potential and rationale of CKD-581 for treatment of MM.

Extraction of Intercellular Material from Rice Leaf Tissue (수도의 세포간극 물질 추출)

  • Park, W.M.;Son, E.R.;Ko, Y.H.;Yoo, Y.J.;Lee, Y.S.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.3
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    • pp.323-327
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    • 1983
  • The intercellular material was extracted from rice leaf tissue. The quantitative tests of protein and soluble carbohydrate, and activity of peroxidase, showed differences between tissue extract and intercellular material. Also electrophoretic patterns of peroxidase and esterase isozymes were not similar between them. It indicated that the intercellular material which was extracted, was not the mixture of cellular materials.

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Electrochemical properties of the mugwort-embedded biosensor for the determination of hydrogen peroxide (쑥을 이용한 과산화수소 정량 바이오센서의 전기화학적 성질)

  • Lee, Beom-Gyu;Park, Sung-Woo;Yoon, Kil-Joong
    • Analytical Science and Technology
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    • v.19 no.1
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    • pp.58-64
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    • 2006
  • A mugwort-tissue-based modified carbon paste electrode was constructed for the amperometric detection of hydrogen peroxide and its electrochemical properties are described. Especially the amperometric signal was very stable and bigger than any other enzyme electrode studied in this lab. The effect of tissue composition on the response was linear within the wide range of experiment and the linearity of Lineweaver-Burk plot showed that the sensing process of the biosensor is by enzymatic catalysis. And pH dependent current profile connoted that two isozymes are active in this system.

Comparison of Glucuronidating Activity of Two Human cDNAs, UDPGTh1 and UDPGTh2

  • Kim, Soon-Sun;Owens, Ida-S.;Sheen, Yhun-Yhong
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.454-458
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    • 1997
  • Two human liver UDP-glucuronosyltransferase cDNA clones, HLUG25 and UDPGTh2 were previously shown to encode isozymes active in the glucuronidation of hyodeoxycholic acid (HDCA) and certain estrogen derivatives (e.g., estriol and 3,4-catechol estrogens), respectively. in this study we have found that the UDPGTh2-encoded isoform (UDPGTh2) and HLUG25-encoded isoform (UDPGThl) have parallel aglycone specificities. When expressed in COS 1 cells, each isoform metabolized three types of dihydroxy- or trihydroxy-substituted ring structures, including the 3,4-catechol estrogen (4-hydroxyestrone), estriol, 17-epiestriol, and HDCA, but the UDPGTh2 isozyme was 100-fold more efficient than UDPGTh1. UDPGTh1 and UDPGTh2 were 86% identical overall (76 differences out of 528 amino acids), including 55 differences in the first 300 amino acids of the amino terminus, a domain which conferred the substrate specificity. The data indicated that a high level of conservation in the amino terminus was not required for the preservation of substrate selectivity. Analysis of glucuronidation activity encoded by UDPGTh1/UDPGTh2 chimeric cDNA constructed at their common restriction sites, Sac I (codon 297), Nco I (codon 385), and Hha I (codon 469), showed that nine amino acids between residues 385 and 469 were important for catalytic efficiency, suggesting that this region represented a domain which was critical for the catalysis but distinct from that responsible for aglycone-selection. These data indicate that UDPGTh2 is a primary isoform responsible for the detoxification of the bile salt intermediate as well as the active estrogen intermediates.

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Analysis of brain protein expression in developing mouse fetus (임신일령에 따른 생쥐 태아 뇌조직의 단백질 발현 양상 분석)

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yun-Fei;Woo, Je-Seok;Jin, Dong-Il
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.65-70
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    • 2011
  • Development of mouse fetus brains can be defined morphologically and functionally by three developmental stages, embryo day (ED) 16, postnatal stage one week and eight weeks. These defined stages of brain development may be closely associated with differential gene expression rates due to limited cellular resources such as energy, space, and free water. Complex patterns of expressed genes and proteins during brain development suggests the changes in relative concentrations of proteins rather than the increase in numbers of new gene products. This study was designed to evaluate early protein expression pattern in mouse fetus brain. The mouse brain proteome of fetus at ED 15.5, and 19.5 was obtained using 2-dimensional gel electrophoresis (DE). Analysis of the 2-DE gels in pH 3-10 range revealed the presence of 15 differentially expressed spots, of which 11 spots were identified to be known proteins following MALDI-TOF analysis; 3 spots were up-regulated and 8 spots were down-regulated in the mouse fetus brain at ED 15.5. UP-regulated proteins were identified as MCG18238, isoform M2 of pyruvate kinase isozymes M1/M2, isoform 2 of heterogeneous nuclear ribonucleoprotein K, heterogeneous nuclear ribonucleoprotein H2, creatine kinase B-type, 40S ribosomal protein SA and hemoglobin subunit beta-H1. Down-regulated proteins were putative uncharacterized protein, lactoylglutathione lyase and secreted acidic cysteine rich glycoprotein. Our results revealed composite profiles of mouse fetus brain proteins related to mouse fetus development by 2-DE analysis implying possible roles of these proteins in neural differentiation.

Isolation of Guaianolides with ACAT Inhibitory Activity from the Leaves and Stems of Chrysanthemum boreale Makino (산국의 잎과 줄기에서 ACAT 저해활성을 가지는 Guaianolides의 분리)

  • Lee, Jong Rok;Park, Moon Ki
    • Journal of Environmental Science International
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    • v.26 no.11
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    • pp.1275-1284
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    • 2017
  • Acyltransferase (AT) catalyzes the transfer of an acyl moiety from acyl-coenzyme A (acyl-CoA) to an acceptor. ATs play important roles in the maintenance of homeostasis in the human body and have been linked to various diseases; therefore, several ATs have been proposed as potential targets for the treatment or prevention of such diseases. The AT family includes acyl-CoA:cholesterol AT (ACAT), diacylglycerol AT, and monoacylglycerol AT for the metabolism of lipids. Furthermore, recent molecular biological studies revealed the existence of their isozymes with distinct functions in the body. ACAT plays a critical role in the formation of cholesteryl esters from cholesterol and fatty acids, and is a potential target for treating hypercholesterolemia. During an experiment designed to discover biologically active compounds from herbal medicines, we isolated two known guaianolide sesquiterpene lactones from Chrysanthemum boreale Makino (Compositae). The lactones were characterized from their spectroscopic data (NMR, IR, MASS). These compounds were subjected to ACAT inhibition assay. Here, we report the isolation and structural elucidation of the compounds 8-o-acetyl-2-methoxy-10-hydroxy-3,11(13)-guaiadiene-12,6-olide and 8-acetyl-3,10-hydroxy-4(15),11(13)-guaiadiene-12,6-olide. In the ACAT inhibition assay, compound 1 showed strong inhibitory activity, with an $IC_{50}$ value $45{\mu}g/mL$, whereas compound 2 did not exhibit significant inhibitory activity with an over $100{\mu}g/mL$.

Genetic Differences between the Tolerant and the Sensitive Trees in an Air Polluted Prunus sargentii Stand (대기오염(大氣汚染) 피해지역(被害地域)의 산벚나무 임분내(林分內) 내성(耐性) 및 감수성(感受性) 개체(個體)의 유전적(遺傳的) 차이(差異))

  • Lee, Seok Woo;Woo, Su Young;Koo, Yeong Bon;Lee, Soung Kyu
    • Journal of Korean Society of Forest Science
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    • v.87 no.1
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    • pp.74-81
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    • 1998
  • The estimates of genetic diversity based on 7 polymorphic loci coding for 6 isozymes were compared between the tolerant and the sensitive trees in an air polluted Prunus sargentii stand located on Ulsan industrial area. Although we could not observe statistically significant differences in allele and genotype frequencies between the two groups except the gene locus Got-2, the unique alleles and genotypes were observed in the tolerant group. All the genetic parameters such as genetic multiplicity, genetic diversity, and heterozygosity revealed that a greater amount of genetic variations existed in the tolerant group.

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Inheritance and Linkage of Some Polymorphic Isozymes in Ginkgo biloba L. (은행나무의 몇가지 다형적(多形的) 동위효소(同位酵素)의 유전양식(遺傳樣式) 및 연관(連關))

  • Kwon, Hae-Yeun;Kim, Zin-Suh
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.527-535
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    • 2000
  • Isozyme variants of 15 enzyme systems were analyzed in megagametophytes of Ginkgo biloba L. Five enzyme systems (ADH, G6PD, IDH, MPI, and UGPP) appeared to be monomorphic. Only 11 isozyme zones observed in 10 enzyme systems were polymorphic : ACON-A, FST-B, GDH-A, GOT-B, MDH-B, MDH-C, MNR-A, PGI-B, PGM-A, 6PGD-B and SKDH-B. The segregation ratio and heterogeneity at most polymorphic zones suggested that each isozyme zone was controlled by a single locus with codominant alleles, but significant deviation from 1 : 1 segregation was observed at MDH-B in pooled data. Three pairs of isozyme loci (ACON-A : MDH-B, GOT-B : PGI-B, and MNR-A : SKDH-B) were found to be weakly linked. Recombination frequencies between them ranged from 0.38 to 0.40 (p<0.05).

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