• Title/Summary/Keyword: Isoenzyme

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Blood chemical research for the diagnosis of cardiac disease in ruminant (반추수 심장질병의 진단을 위한 혈액화학적 연구)

  • Kim, Duck-hwan;Lee, Kyo-young;Chang, Seuk-jin;Song, Kun-ho;Lee, Yoon-kyung;Yoon, Sang-bo
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.399-404
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    • 1995
  • In order to clarify the significance of serum CPK for the diagnosis of the cardiac disease in ruminant, the fluctuation of serum total CPK activities and CPK isoenzyme fractions was examined before and after operation in Korean native goats with artificially induced cardiac injury. In the change of serum total CPK activities, those of experimental group were changed with higher values than those of control group and significant increase was found on 2-week after operation(P<0.05). In the fluctuation of serum CPK isoenzyme fractions, significant increase of both CK1 and CK3 fractions was observed on 2-week after operation in experimental group(P<0.05), but significant difference was not found in CK2 fraction between experimental and control groups. From these findings, it is considered that taking note of increase of not only serum total CPK activities but also CK1 and CK3 fractions is important for the diagnosis of traumatic cardiac disease in ruminant.

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Biochemical and molecular characterization of a strain KA/S2 of Acnnthamoebc castellanii isolated from Korean soil (카스텔란니가시아메바(Acanthamoeba castellanii) 한국 토양분리주 KA/S2의 생화학적 및 분자생물학적 특성)

  • 정동일;공현희
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.79-86
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    • 1996
  • A strain, KA/S2, isolated from Korean soil and morphologically assigned to Acanthcmoebc cQsteLlcnii, was characterized by isoenzyme analysis , and total proteins profile, End mitochondrial (Mt) DNA restriction fragment length polymorphism (RFLP) , and compared with four reference strains assigned to the species (the authenitic Castellani, Neff, Ma, and Chang strains). It was found that four isoenzyme, total proteins, and Mt DNA RFLP patterns by eight restriction endonucleases of the strain KA/S2 were identical with those of the Neff strain, isolated from soil of California, USA. The Chang strain was unique in its morphology and total protein patterns. Interstrain polymorphisms of isoensyme profiles and Mt DNA RFLP patterns were observed among the Castellani, Neff, Ma, and Chang strains. Mt DNA RFLP was confirmed to be highly appropriate for the strain characterization and identification of Acnnthamoeba spry.

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Expression of protein kinase C in the testes of horse (말 정소내 protein kinase C의 발현)

  • Jin, Jae-kwang;Shin, Tae-kyun
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.9-15
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    • 1998
  • To investigate the involvement of protein kinase C(PKC) isoenzyme in the testes which control spermatogenesis and hormone secretion, we examined cellular distribution of four types of PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$ in the horse testes using PKC antisera by western blot analysis and immunohistochemistry. By the western blot analysis, PKC $\alpha$ and ${\beta}I$ were detected at 82KD, while PKC ${\delta}$ and ${\theta}$ were detected at 80KD in the testes of both juvenile and adult horses. In juvenile horse, PKC $\alpha$, ${\delta}$ and ${\theta}$ except ${\beta}I$ were not detected in the cells of the testes, whereas PKC ${\beta}I$ was immunoreacted with only in spermatocytes. In adult, PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$isoenzymes were localized in interstitial cells of the testes. In the seminiferous tubules, PKC ${\beta}I$ is localized in spermatocyte, spermatid and spermatozoa, while PKC ${\delta}$ is localized only in spermatids. We suggest that this is a first report to localize PKC in the testes of horse and PKC isoenzymes are upregulated in the cells of horse testes depending on ages. These findings also suggest that certain PKC isoenzyme plays an important role in the signal transduction of spermatogenic cells and interstitial cells in horse testes.

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The Characteristics of I269S and I224S Double Mutant Horse Liver Alcohol Dehydrogenase (I269S와 I224S 이중변이 알코올 탈수소효소의 특성)

  • Ryu, Ji-Won;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.756-764
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    • 1997
  • Ile-224 in I269S mutant horse liver alcohol dehydrogenase isoenzyme S (HLADH-S) was mutated to serine by site-directed mutagenesis in order to study the role of the residue in c oenzyme binding to the enzyme. The specific activity of the I269S and I224S mutant enzyme to ethanol was increased 6-fold and all Michaelis constants($K_a,\;K_b,\;K_p,\;and\;K_q$,/TEX>) were larger than those for the wild-type and I269S enzyme. The substitution decreased the afffinity to coenzymes and increased the specific activity of the enzyme. The mutant enzyme showed the highest catalytic efficiency for octanol among the primary alcohols. But it didn`t have activities on retinoids and 5${\beta}$-cholanic acid-3-one. From these results, it was confirmed that the hydrophobic interaction of Ile-224 residue with coenzyme was related to coenzyme affinity in ADH reaction. The substitution also affected the substrate affinities to the enzyme.

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Expression of recombinant plasmids harboring glucoamylase gene STA in saccharomyces cerevisiae (Glucoamylase 유전자 STA를 포함한 재조합 플라스미드들의 saccharomyces cerevisiae에서의 발현)

  • 박장서;박용준;이영호;강현삼;백운화
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.181-187
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    • 1990
  • STA gene coding glucoamylase was introduced into haploid Saccharomyces cerevisiae SHY3 and polyploid Saccharomyces cerevisiae 54. We constructed the recombinant plasmid by substituting the promoter region of alcohol dehydrogenase isoenzyme I gene for that of STA gene to increase the expression of STA gene and found that the activity of glucoamylase was increased in transformants. The plasmid stability was improved remarkably when we got the STA gene into the plasmid which had centromere. The activity of glucoamylase and transformation frequency of it, however, was decreased because of low copy number. Industrial polyploid strain was transformed with the recombinant plasmid having the $2\mu$ origin of replication and STA gene. It produced more alcohol than host when fermented in liquefied starch media. The industrial strain, however, was not transformed with the autonomously replicating plasmid containing centromere.

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Studies on the Antioxioative Character in the Etnyl Acetate Extractions of Rumex crispus (Rumex crispus의 에칠아세테이트 추출물의 항산화 성분에 관한 연구)

  • 신춘혜
    • KSBB Journal
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    • v.16 no.6
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    • pp.592-602
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    • 2001
  • This study was undertaken to investigate the antioxidative substance and activity of ethyl acetate extracted from Rumex crispus. Sample extracted follow in proper course of a solvent. Material refinement was carried out using silicagel column and Sephadex LH-20 column chromatography. Material sorting was carried by Gas Chromatography(GC/MS). 1,1-Diphenyl-2-Picrylhydrazyl(DPPH) free radical scavenging and enzyme activity were measured for antioxidative activity. as result of testing by DPPH free radical scavenging activity, Antioxidative activity was shown as the highest in the root, then leaf and stem in order. Ethyl acetate extraction of root part were 50% inhibitory concentration (IC50) Rumex activty(6.1 ug/mL). Rumex nipponicus(9.8 ug/ml) and Rumex acetoceae(31.5 ug/mL) in leaf part. The highest antioxidative activity of sample refined through silicagel column chromatography of Rumex crispus was appealed Fraction 5(IC50;3.57 ug/mL) in root and Fraction 6(IC50;85.9 ug/mL) in leaf. Fraction 5 in roof & Fraction 6 in leaf were refined using Sephadex LH-20 column chromatography. The highest antioxidative activity were appeared Fraction 4 (IC50;3.57 ug/mL) and Fraction 4 (IC50;18.41 ug/mL)in leaf. As for main phenol compounds 2,6-Dichloro-4-nitropnenol and 2-Isopropyl-5-methyl Phenol were identified in root and leaf, While 4-Vinyl-2- methoxy-phenol and 2,3-Dihydro- benzofuran were identifica ted only in leaf. Enzyme activity was shown low both in peroxidase(PDD) Non-activate(IU/mg protein)and in Superoxide dismutase(SOD) non-activate(IU/mg protein). 2,6-Dichloro-4-nitrophenol, 2-Isopropyl-5-methyl phenol, 4-Vinyl-2-methoxy-phenol were obtained in this experiment and these compounds are phenolic compounds which have OH group in the structure. With the result of this study these phenolic compounds which are extracted from Rumex crispus have high antioxidative effect. This antioxidative effect of Rumex crispus can be applied for chromo-preventive and antioxidative supplements which can be used for anti-allegy, aging, anti-tumor, aging and other oxidative disease for health promotion.

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Electrophoretic Studies of Peroxidase Isoenzymes for Ages, Locations, Organs, Cultivars and Hybrids in the Apple Trees (사과의 수령(樹齡), 지역(地域) 및 부위(部位)에 따른 Peroxidase 동위효소(同位酵素)의 변동(變動)과 품종간(品種間) 및 그 교잡묘간(交雜苗間)의 차이(差異))

  • Chung, Keun Ok;Kim, Young Rae
    • Korean Journal of Agricultural Science
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    • v.16 no.1
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    • pp.26-35
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    • 1989
  • Studies were carried out to compare from the viewpoint of peroxidase isoenzyme patterns by ages, locations and organs. In addition, the relationships among six cultivars peroxidase isoenzyme patterns were compared and the relationships between cultivars and their hybrids were examined. The results are summarized as follows. 1. The zymogram was not affected by the ages of 'Fuji' cultivar. 2. There was no locational difference in zymogram among Suwon, Taejon and Yesan in 'Fuji' and 'Tsugaru' cultivars. 3. Significant difference in isozyme patterns was found between stems and leaves in 'Fuji' and 'Tsugaru' cultivars. 4. There were some differences in zymogram among 'Ralls Genet', 'Fuji', 'Jonathan', 'Starking', 'Golden Delicous' and 'Tsugaru' cultivars. 5. Classification was made by the peroxidase isozyme development between cultivars and their hybrids. 1) The type showed both parent's isozyme pattern. 2) The type showed a loss of a part of parent's isozyme bands. 3) The type developed new isozymes which were not detected in parent plants.

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Detection of Protein Kinase C Isoenzymes in the Growth of Human Epidermal Keratinocytes by Growth Factors (Growth Factor를 처리한 피부상피세포로부터 Protein Kinase C Isoenzyme의 검출)

  • Eun-Young Joo;Nam-Woo Kim
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.83-91
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    • 2000
  • Subconfluent neonatal human epidermal keratinocytes were treated with a concentration 200 ng/$m\ell$ of human recombinant epidermal growth factor (hrEGF), human recombinant insulin-like growth factor-1 (hrIGF-1), and with a combination of hrEGF and hrIGF-1. Cytoplasmic and membrane-associated proteins were extracted and assayed. Proteins were separated by SDS-PAGE, and subjected to the western blot analysis. In the cytoplasmic fraction, the PKC concentration of keratinocyte treated with hrIGF-1 was higher than the control group, but the concentration of control group was the highest than the others in the membrane fraction. In the cytoplasmic fraction, EGF stimulated PKC-$\beta$II, -$\delta$, -$\theta$, and also stimulated PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$ and -$\theta$ in the membrane fraction. IGF-1 stimulated PKC-$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic, PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$, - $\varepsilon$ and -$\theta$ in the membrane. In the cells treated with a combination of EGF and IGF-1, PKC-$\alpha$, -$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic fraction, PKC-$\alpha$, -$\delta$, -$\Im$, -$\varepsilon$ and -$\theta$ in the membrane fraction were stimulated.

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Effects of Temperature Stress and Paraquat on SOD Activity and Photochemical Efficiency of PSII in Leaves of Araliaceae Plants (두릅나무과 식물의 SOD 활성과 광계II의 광화학적 효율에 미치는 온도 스트레스와 Paraquat의 영향)

  • 오순자;고정군;김응식;오문유;고석찬
    • Korean Journal of Environmental Biology
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    • v.17 no.2
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    • pp.199-204
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    • 1999
  • The effects of temperature stress and paraquat on the superoxide dismutase (SOD) activity and the photochemical efficiency of photosystem II were studied in the leaves of Araliaceae plants. The SOD activity of Acanthopanax koreanum leaf discs increased at 4$^{\circ}C$ and 28$^{\circ}C$, and increased significantly at 4$^{\circ}C$ ,28$^{\circ}C$ and 35$^{\circ}C$ in the presence of paraquat. However, the SOD activity of Dendropanax morbifera leaf discs decreased at 4$^{\circ}C$, 28$^{\circ}C$ and 35$^{\circ}C$ regardless of paraquat treatment. The photochemical efficiency of photosystem II, Fv/Fm, of leaf discs of A. koreanum and D. morbifera fell remarkably at 35$^{\circ}C$. In the presence of paraquat, the Ev/Fm values fell slightly at 4$^{\circ}C$ in A. koreanum leaf discs and at 35$^{\circ}C$, in D. morbifera leaf discs. These results indicate that A. koreanum plants are more resistant to temperature stress or oxidative stress than D. morbifera plants although their photochemical efficiency falls slightly at 4$^{\circ}C$ in the presence of paraquat.

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Characterization of Thermolabile Pectinesterase and Thermostable Pectinesterase Separated from Valencia Orange (Valencia 오렌지로부터 분리 정제한 비내열성 및 내열성 Pectinesterase의 성질)

  • Hou, Won-Nyoung;M.R., Marshall
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.666-672
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    • 1995
  • This study was carried out to characterize thermolabile pectinesterase (TLPE) and thermostable pectinesterase (TSPE) separated from crude PE of Valencia orange in order to investigate the preventive measures of cloudy juice clarification. The TLPE was observed to be mixture of several isoenzymes with the same molecular weight of 36 KD (37.5 KD) but different isoelectric point of pH 8.4, 8.7, 8.9, 9.8 and ${\geq}10$ which were unstable at $70^{\circ}C$, and the TSPE also was found to be mixture of two or three isoenzymes with the same molecular weight of 53 KD (50 KD) but different isoelectric point of pH 8.7, 9.2 and ${\geq}10$ which had slightly different stability from one another at $70^{\circ}C$. The TLPE and the TSPE had the optimum reaction pH of 7.0 and $7.0{\sim}8.5,\;appK_{M}$ of 1.1 and 1.7 mg/ml, appVmax of 0.53 and $1.01\;{\mu}mol/min/{\mu}g$, and the turnover number of 19.000 and 54,000 mol/mol/min toward Kodak pectin, respectively. The TSPE had higher storage stabiblity and cloud loss effect on orange juice than the TLPE. Above all, the crude PE was most effective on orange juice cloud loss among the PEs used.

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