• Title/Summary/Keyword: Isoenzyme

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Subcellular Distribution and Characteristics of Polyphenol Oxidase from Perilla frutescens Leaves (들깨잎 polyphenol oxidase의 세포내 분포 및 특성)

  • 김안근;김유경
    • YAKHAK HOEJI
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    • v.43 no.6
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    • pp.709-715
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    • 1999
  • Polyphenol oxidase (PPO) activity in 200×g (cell wall), 4,000×g (plastid), 100,000×g (mitochondrial) and soluble fractions of the perilla leaves was monitored in the upper, middle and lower sections of the plant. In the course of plant growth, PPO activities in plastid and mitochondrial fractions were decreased, while those in cell wall fraction were maintained. During growing process, specific activities and PPO activities of each fraction were decreased, while total phenol content were decreased in middle (middle) and then increased in later stage (lower). Cell wall, plastid, mitochondrial (pellet) and soluble fraction had slightly different pH optima and substrate specificities. Isoenzyme patterns were identical in two bands for PPO activity in different subcellular fractions. Their molecular weights were 37KD and 48KD respectively.

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Comparision of Polypeptide Patterns by 2-D PAGE in Fusarium Species (이차 전기영동법을 이용한 Fusarium 속의 다당류 비교)

  • Min, Byung-Re
    • The Korean Journal of Mycology
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    • v.23 no.4 s.75
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    • pp.359-368
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    • 1995
  • F. napiforme, F. beomiforme and F. nygamai could not be classified in any of the existing sections of the genus Fusarium. To discuss of the exact taxonomic relationships among these species, the cellular polypeptide patterns were compared by using 2-D PAGE. Polypeptide pattern of F. beomiforme was different from those of other two species and was more similar to F. oxysporum in section Elegans. F. nygamai and F. napiforme might be another same section which would lie between section Liseola and section Elegans. The results were consistent with the comparison of isoenzyme patterns in these species.

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The Role of Lys-228 Residue in Horse Liver Alcohol Dehy-drogenase Activity

  • Cho, Sun-Hyoung;Ryu, Ji-Won;Lee, Kang-Man
    • Archives of Pharmacal Research
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    • v.18 no.2
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    • pp.100-104
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    • 1995
  • Lys-228 in horse liver alcohol dehydrogenase isoenzyme E(HLADH-E) was mutated to glycineby site-directed mutagenesis. The specific activity of the mutant enzyme was increased about 4-fold nad Michaelis constants for $NAD^+(K_a){\;}and{\;}NADH(K_q)$ increased by about 350-and 50-fold, respectively. The wild-type enzyme and K228TG mutant enzyme were treated with ethylacetimidate. Acetimidylation of the wild-type enzyme increased the activity about 10-fold, but the mutant enzyme ws little affected. These results confirm that Lys-228 residue plays an important role in the activity of the enzyme through forming the hydrogen bond with adenosine ribose of $NAD^+$.

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Analysis of risk factors of atrial fibrillation after coronary artery bypass grafting (관상동맥 우회로 조성수술후 발생하는 심 방세동의 위험요인 분석)

  • Yu, Gyeong-Jong;Go, Yeong-Ho;Im, Sang-Hyeon;Gang, Myeon-Sik
    • Journal of Chest Surgery
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    • v.29 no.6
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    • pp.599-605
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    • 1996
  • A total of 249 patients undergoing isolated coronary revascularization were studied for the occurrence of postoperative atrial fibrillation(AF). Possible associations of this arrhythmia with various preoperative, intraoperative and postoperative factors were studied by univariate and multivariate analysis. The overall incidence of postoperative AF was 15%, with the median time occurence of 48 hours(mean time : 59.1 $\pm$ 56.9 hours) after arrival to the intensive care unit. Cardiac index decreased significantly after occurence of AF(p=0.001). There were no in-hospital complications in those patients with AF. Univariate studies indicated preoperative ejection fract on(EF), triglyceride level, postoperative peak CKMB isoenzpme and atrial pacing to be the dominant factor promoting postoperative AF, with an increasing prevalence in lower EF(p=0.025), triglyceride(p=0.006) and peak CKMB isoenzyme(p=0.002), and in patients with atrial pacing(p=0.001). Hospital stay(p=0.001) and late mortality(p=0.003) were significantly increased in patients with postoperative AF Multivariate analysis showed that body weight and postoperative atrial pacing to be the dominant factor promoting postoperative AF, with an increasing prevalence in over- weight patients(p=0.011) and patients with atrial pacing(p=0.001). Both univariate and multivariate analy- sis showed that the age was not a significant factor but tended to promote postoperative AF respectively (p=0.053, 0.064). After 30.1 $\pm$ 11.4 months gfollow-up, those patients with AF had sinus rhythm. We think that we must try to prevent postoperative AF after ccoronary artery bypass grafting because of its deleterio s hemodynamic effect, prolonged hospital stay, and increased late mortality.

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The characteristics of selected trees of putative hybrid pines between Pinus thunbergii and Pinus densiflora (곰솔과 소나무의 자연잡종(自然雜種)으로 추정(推定)되는 잡종(雜種)소나무의 특성(特性))

  • Son, Doo Sik;Kwon, Chil Yong;Park, Sang Jun
    • Journal of Korean Society of Forest Science
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    • v.79 no.2
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    • pp.127-137
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    • 1990
  • This study was carried out to investigate some characteristics of selected trees recognized as natural hybrid between Pinus densiflora and Pinus thunbergii for improving resistance to pine leaf gall midge. The results obtained were as follows. l. In the mean annual increment and the growth in the period of latest 5 years of DBH, hybrid pines showed intermediate value of the parents, Pinus densiflora and Pinus thunbergii. However, there was remarkable variation among individuals of hybrid pine. 2. The trees having resin duct index(RDI) value larger than 0.1 for the position of resin duct in needle was classified as hybrid pine. The leaf and leaf sheath of Pinus thunbergii was longer than that of Pinus densiflora and the distance between rows of stomata on needles of Pinus thunbergii was longer than that of Pinus detasiflora, and the values of those observations on hybrid pine were intermediate between the parent species. However, differences among individual was observed. 3. In the size of cone and seed, and weight of 1000 seeds, like in the leaf characteristics, Pinus thunbergii showed higher value than Pinus densiflora and those values of hybrid were intermediate between the parent species. 4. Assuming isoenzyme ADH-$B_2$, ME-$A_2$, PGI-$B_1$, and PGI-$B_2$ alleles observed in the hybrids were introgressed from Pinus thunbergii, the hybrid pine can be easily identified by these isoenzyme alleles. However, the individuals which do not have those alleles can not be identified.

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Changes of Reserved Substance and the Peroxidase Activity in Tomato Fruits during the Storage of Sub-atmosphere Pressure (감압저장중 도마도 과실의 저장물질과 Peroxidase 활성의 변화)

  • Sohn, Tae Hwa;Choi, Jong Uck;Cheon, Seong Ho
    • Current Research on Agriculture and Life Sciences
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    • v.3
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    • pp.92-98
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    • 1985
  • The experiments were conducted to investigate the activity changes of peroxidase, existence of isoenzyme and the changes of reserved substances in tomato under subatmospheric pressure storage condition. The results obtained were as follows: 1. Soluble fraction showed the highest peroxidase activity and followed by cell wall fraction, mitochondria fraction and ribosome fraction in that order. 2. Peroxidase activity was decreased during the ripening and senescence period in tomato. Especially, peroxidase activity in tomato was higher at a room temperature ($25^{\circ}C$) than at a low temperature ($15^{\circ}C$). The decreasing inclination was similar in both treatment. The peroxidase activity was higher in 380 Torr, than in 570 Torr. 3. At least, two isoperoxidases(Soluble or solubilized) were identified from different extraction procedures. Three of four isoenzymes were recognized from a vertical slab of polyacrylamide gel electrophoresis. 4. The changes of components in tomato under SAP were generally affected by temperature and pressure. Especially, quality of tomato stored at a low temperature ($15^{\circ}C$) and SAP (380 Torr.) was best during storage.

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Alterations of Glutathione Peroxidase Patterns by Stressor Treatment in Rice Seedling Roots (스트레스 물질에 의한 벼 glutathione peroxidase 활성패턴 변화)

  • Kim, Yoon-Kyoung;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.53-59
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    • 2005
  • The effect of various stressors such as reductant ascorbic acid, signalling molecules (salicylic acid and methyl jasmonic acid), heavy metals $(NiCl_2,\;and\;MnSO_4)$ and NaCl on the glutathione peroxidase (GPX) activities and isoenzyme expression patterns were investigated in rice seedling roots. Total GPX activity increased according to the increase of ascorbic acid concentration. Prominent enhancement of GPX1 isozyme due to ascorbic acid contributed to the increase of total GPX activity. GPX showed different reactivity toward salicylic acid and methyl jasmonic acid. GPX activity increased at 0.1 mM salicylic acid, and then decreased thereafter. However, GPX increased gradually in a methyl jasmonic acid concentration-dependent manner, and 3 fold increase of GPX activity was found at 1 mM methyl jasmonic acid. Moreover, GPX1 isozyme increased according to the increase of salicylic acid, while GPX1 isozyme decreased according to the increase of methyl jasmonic acid. When metal ions were treated, GPX activity increased considerably according to the increase of $NiCl_2$ concentration, however, GPX activity increased about 2 fold at 0.5 mM $CuSO_4$ and then decreased. Enhancement of GPX1 isozyme contributed to the increase of total GPX activities in $NiCl_2-treated$ and $MnSO_4-treated$ rice seedlings. Total GPX activity increased 1.7 fold in response to 300 mM NaCl. Especially GPX2 isozyme showed gradual increase according to the increase of NaCl concentration.

Restriction endonuclease analysis of mitochondrial DNA of Acanthamoebn sp. YM-4 (Korean isolate) (Acanrhamoeba sp. YM-4의 미토콘드리아 DNA의 RFLP분석)

  • Sin, Ho-Jun;Im, Gyeong-Il;Jeon, Gwang-U
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.119-126
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    • 1997
  • Acanthnmoebn sp. YM-4 is simitar to A. culbertsoni based upon morphological characteristics of trophozoites and cysts. However, based on other characteristics, pathogenicity to mice, in uitro cytotoxicity and isoenzyme patterns, Acanthomoebo sp. YM- 4 was quite different from A. culbertsoni. Restriction fragment length polymorphism (RFLP) analysis of mtDNA is useful in the classification of members belonging to the genus Acanthcmoebn. Therefore, in this study, RFLP analysis of Acnnthcmoeba mtDNAs was accomplished using five restriction enzymes: Hnelll, Hinull, Clcl, Pudl and ScE. Each restriction enzyme produced approximately 3-15 fragments (range: from 0:6 kip to 34.4 kbp) . The mtDNA genome size, calculated by the summation of restriction fragments, averaged 46.4 kbp in Acnnthamoeba sp. YM-4,48.3 kbp in A. culbertsoni and 48.8 kbp in A. polyphaic, respectively. Digested mtDNA fragments of Accnthcmoeba sp. YM-4 contained nine and seven same size fragments, respectively, from a total of 67 and 69 fragments observed in A. culbertsoni and A. polyphcgn. An estimate of the genetic divergence was 10.1% between Acanthamoebc sp. YM-4 and A. culbertsoni, and 9.9% between Acanthamoebn sp. YM-4 and A. polyphcga.

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Properties of Chorismate Mutase from intrasporangium sp. (Intrasporangium속 방선균의 Chorismate Mutase 성질)

  • 조원대;신광순;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.310-315
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    • 1988
  • Two isoenzymes of chorismate mutase(E.C.5.4.99.5) designated as chorismate mutase I(CM I) and chorismate mutase II(CM II), were detected and partially purified from a sp. of intrasporangium isolated from soil. CM I and CM II had pH optima of pH 6.5 and 8.0, respectively and showed the same temperature optimum of 45$^{\circ}C$. The activation energy of the enzymatic reaction was estimated to be 14.7kcal/ mole with CM I and 10.8kcal/mole with CM II. The affinity of isoenzyme CM I for substrate(Km= 1.35mM) was almost the same level as that of CM II(Km = 1.22mM). Both isoenzymes were stable at pH values ranged from pH 6.5 to 9.0, but rapidly denaturated at temperatures above 45$^{\circ}C$. CM II was activated about 7$^{\circ}C$ of its activity by $Ba^{++}$ or $Mg^{++}$ while CM I was slightly inhibited by the same metal ions. Thiol compounds were found not to be necessary for stability of the two enzymes but Co$^{++}$ and EDTA had a little stabilizing effect on CM II only. p-Chloromercuribenzoate strongly inactivated the activities of both enzymes but the reducing agents such as dithiothreitol and L-cysteine protected them against the pCMB inhibition.

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Cloning and Sequencing of the pelCl Gene Encoding Pectate Lyase of Erwinia carotovora subsp. carotovora LY34 (Erwinia carotovora subsp. carotovora LY34에서 pelCI 유전자 클로닝)

  • Lim, Sun-Tech;Park, Yong-Woo;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.380-387
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    • 1997
  • Phytopathogenic Erwinia carotovora subsp. carotovora (Ecc) LY34 causes plant tissue maceration by secretion of pectinolytic enzymes such as pectate Iyase (PL) existed as multiple isoenzyme form. Genomic DNA from Ecc LY34 was digested with Sau3Al and ligated into the BamHI site of pBluescript ll $SK^+$. Among them, a clone hydrolyzing polypectate was selected and its DNA was digested with BamHI. Through the subsequent subcloning the resulting 3.1 kb fragment, corresponding to a peICI, was subcloned into pLYPA 100. The structural organization of a peICI gene encoding a 374 amino acid residues consists of an open reading frame (ORF) of 1,122 bp commencing with a ATG start codon and followed by a TAA stop codon. PeICI contained a typical prokaryotic signal peptide of 22-amino acid. Since the deduced amino acid sequences of PeICl protein was very similar to those of PelIII of Erwinia carotovora subsp. carotovora, and to those of Pel3 of Erwinia carotovora subsp. atroseptica, and to those of PeIC of Erwinia carotovora subsp. carotovora, it belong to the same family PLbc group. The 374-amino acld PeICI had a calculated Mr of 40,507 and pI of 7.60.

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