• 제목/요약/키워드: Isoenzymes

검색결과 90건 처리시간 0.029초

인삼 근절편 배양시 Peroxidase Isozyme에 관한 연구 (Peroxidase Isozyme in Root Differentiation from Cultured Ginseng Root Explants)

  • 김명원
    • Journal of Plant Biology
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    • 제29권4호
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    • pp.233-242
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    • 1986
  • In order to pursue some physiological studies on organogenesis in ginseng tissue culture, ginseng root explants were cultured on a modified MS medium containing NAA and kinetin. The activities of peroxidase and some enzymes were investigated and their isoenzyme patterns were also observed. The activity of peroxidase decreased by 20% in one week's culture and increased thereafter by 80% in culturing for 7 weeks compared with the control group. Glucose-6-phosphate dehydrogenase activity increased by 400% after culturing for 5 weeks and increased during the days preceeding root formation. The activities of glutamate dehydrogenase and acid phosphatase also increased during the culture. After 3 weeks' culture, new peroxidase isozyme (pH 7.6) appeared and 7 weeks' culture, another new peroxidase isozyme (pH unidentified) appeared. These patterns were also identified by using FPLC. After 7 weeks' culture, a new esterase isozyme of pH 8.5 appeared and isozyme patterns of acid phosphatase were quite changed compared with the isozyme patterns of tissue cultured for 5 weeks. In so far as these new isoenzymes appear distinctively after 7 weeks' culture, root differentiation is supposed to be induced after 7 weeks' culture.

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고려인삼(Panax ginseng C.A.Meyer) 조직배양에서 2.4-dichloro phenoxyacetic acid와 kinetin첨가가 Isoperoxidase 변이에 미치는 영향 (Effect of 2,4-Dichlorophenoxyacetic Acid and Kinetin on Peroxidase Isoenzymes in Ginseng(Panax ginseng C. A. Meyer) Callus Cultures)

  • 김명원;강영희
    • Journal of Ginseng Research
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    • 제7권1호
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    • pp.52-62
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    • 1983
  • This study was undertaken to investigate the influence of kinetin and 2, 4-dichlorophenoxyacetic acid on the rate of growth, the contents of RNA, DNA, and protein. And also the effect of plant growth regulator on isoperoxidases in callus derived from root (root-callus) and petiole (petiolecallus) was investigated. The rate of growth in petiole-callus was higher than the rootcallus at 0.1 mg/l kinetin and 1mgfl 2,4-D. At 1mgll kinetic, the rate of growth increased, but at high concentration the rate of growth decreased fast. The contents of RNA, DNA and protein also increased, but it did not coincide with the increase of the growth rate of callus. The isoperoxidases of callus grown at various amounts of 2,4-D and kinetic occurred in an almost fashion, but those of root-callus appeared different from those of petiole-callus.

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소나무 및 곰솔의 수지구지수(樹脂溝脂數)에 따른 침엽(針葉), 구과(毬果) 및 종자(種子)의 형태적(形態的) 특성(特性)과 동위효소(同位酵素)의 변이(變異) (The Variation of Isoenzymes and Morphological Characteristics of Needle, Cone and Seed According to Resin Duct Index in Pinus densiflora and Pinus thunbergii)

  • 손두식;박상준;황재우
    • 한국산림과학회지
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    • 제79권4호
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    • pp.424-430
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    • 1990
  • 소나무, 잡종(雜種)소나무 및 곰솔의 수지구지수(樹脂溝指數)의 변화(變化)에 따라 침엽(針葉), 구과(毬果) 및 종자(種子)의 형태적(形態的) 특성(特性)과 동위효소(同位酵素) ADH, ME, PGI의 대립유전자빈도(對立遺傳子頻度)의 변이(變異)를 조사(調査)한바 다음과 같다. 1. 수지구지수(樹脂溝指數)가 증가(增加)암에 따라 침엽(針葉)의 길이, 엽초장, 구과(毬果)의 크기, 종자(種子)의 크기, 종자(種子)날개의 크기, 종자(種子) 1000립중(粒重)은 증가(增加)하고 침엽(針葉)의 기공열수(氣孔列數)는 감소하는 경향을 나타내고 있다. 2. 침엽(針葉), 구과(毬果) 및 종자(種子)의 형태적(形態的) 특성(特性)을 판별분석(判別分析) 결과(結果) 수지구지수(樹脂溝指數)와 대채로 일치(一致)하는 경향을 보이고 있으나 개체(個體) 따라 일치(一致)하지 않는 개체(個體)도 있다. 소나무 및 잡종(雜種)소나무에서는 10-25% 정도는 일치(一致)하지 않았고 곰솔에서는 거의 일치(一致)하였다. 3. 각(各) 형태적(形態的) 특성(特性)에 의한 정준판별함수(正準判別函數)의 결과 소나무, 이입교잡종(移入交雜種), 잡종(雜種)소나무, 곰솔의 양적형질중(量的形質中) 수지구지수(樹脂溝指數), 종자(種子)의 1000립중(粒重), 구과(毬果)의 크기 및 엽초장이 각수종(各樹種)의 특성(特性)을 가장 잘 나타내고 있다. 4. 수지구지수(樹脂溝指數)가 증가(增加)함에 따라 잡종도지수(雜種度指數)도 증가(增加)하는 경향을 나타내고 잡종도지수(雜種度指數)와 판별분석(判別分析)의 결과(結果)는 거의 같은 경향을 나타내고 있다. 5. 수지구지수(樹脂溝指數) 증가(增加)함에 따라 동위효소(同位酵素) ADH-$B_2$, ME-$A_2$, 및 PGI-$B_1$, $B_2$ 대립유전자(對立遺傳子)의 빈도(頻度)는 증가(增加)하고 ADH-$B_3$, ME-$A_4$, PGI-$B_3$는 감소하는 경향을 나타내고 있다. ADH-$B_2$, ME-$A_2$ 및 PGI-$B_1$, $B_2$의 인자(因子)는 곰솔에서 소나무로 유입(流入)된 것으로 간주되며 이러한 인자(因子)가 유입(流入)됨에 따라 ADH-$B_3$, ME-$A_4$, 및 PGI-$B_3$는 감소하는 것으로 생각된다.

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정상 및 병적체액중 동종효소분획에 관한 연구 (Study on the Pattern of Isoenzymes in Pancreatic Juice, Serum and Saliva of Rabbit)

  • 김원준;김혜영;이향우;홍사석
    • 대한약리학회지
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    • 제16권2호
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    • pp.15-24
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    • 1980
  • [${\alpha}$]-Amylase catalyses the hydrolysis of starch, glycogen, and related poly- and oligosac-charide by random cleavage of ${\alpha}$-D-(l-4) glucan linkage. In man large amounts of amylase are secreted into the digestive tract by the salivary and exocrine pancreatic gland, minimal amount being produced also in other tissues. It has been known that ${\alpha}$-amylase exists in multiple molecular forms, isoenzyme which can be separated from each other because of difference in their physicochemical properties. By using various methods, several groups of investigator have separated the many isoenzyme in serum, saliva and pancreatic juice. Furthermore, changes of the normal serum isoenzyme pattern is diagnostically useful even when the total serum enzyme activity is noninformative, such as the clinical use of isoenzyme of serum lactate dehydrogenase. Procarboxypeptidase-A which is one of the pancreatic enzymes is also present as isoenzymes. Four forms of procarboxypeptidase-A haye been found in the bovine enzyme and three forms of the porcine enzyme. In human pancreatic juice four forms of procarboxypeptidase-A isoenzyme were found by isoelectric focusing method. Recently, the so-called isoamylase analysis was developed for the diagnostic use of amylase in pancreatic diseases. In alcohotic patients, the serum concentration of pancreatic isoamylase is subnormal and this lowered activity provides strong evidence for pancreatic exocrine insufficiency. The purpose of this study was to elucidate the variations of the isoenzyme of amylase and procarboxypeptidase-A in serum, saliva and pancreatic juice of the experimental animals. The results are as follow. 1) Three main forms of isoenzyme of amylase by isoelectric focusing were found in pancreatic juice of normal rabbit. However, many new bands were appeared in the pancreatic juice of cholic acid administered animal intravenously while the infusion of cholic acid or elastase into pancreatic duct produced the decrease of number of the fractions on the isoelectric focusing. In the case of serum isoenzyme from normal animal, two major and a few minor isoamylases were observed. By injecting alcohol intravenousely the fractions of serum isoamylase were significantly decreased and in contrary to the pattern in the pancreatic juice the infusion of cholic acid or elastase into pancreatic duct exhitited a significant decrease of the isoenzyme of amylase fractions. In saliva from normal animal three main isoamylase were produced of the administration of alcohol. 2) In the case of procarboxypeptidase-A isoenzyme, two major fractions which have isoelectric point at 6.2 and 6.4 and other two minor bands were observed in the pancreatic juice of normal rabbit. By the treatment of the juice with trypsin, only one band was produced on the isoelectric focusing. No procarboxypeptidase was appeared on the electrofocusing by the infusion of cholic acid or phospholipase A into the pancreatic duct of rabbit. However, a single major fraction of procarboxypeptidase-A was appeared at 3 hr after simple ligation of the pancreatic duct. No significant changes were observed in the juice of the alcohol or cholic acid administered group.

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DMH에 의한 비정상적인 혈관 내피세포의 증식에서 Protein Kinase C 동종효소 Alpha 단백발현의 특성 (Characterization of the Expression of PKCα(Isoform) in DMH-induced Vascular Endothelial Proliferation)

  • 남수봉;배용찬;박숙영;최수종
    • Archives of Plastic Surgery
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    • 제34권6호
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    • pp.679-684
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    • 2007
  • Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

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

  • 조원대;신광순;최용진;양한철
    • 한국미생물·생명공학회지
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    • 제16권4호
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    • pp.310-315
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    • 1988
  • Intrasporangium속 방선균의 chorismate mutase 는 CM I과 CMII의 두 isoenzyme으로 존재하며 최적 pH는 각각 6.5와 8.0이며 최적온도는 다같이 45$^{\circ}C$ 부근이었다. 활성화에너지는 CM I이 14.7 Kcal/mole과 CMII가 10.8kcal/mole로 계산되었으며 각각 1.35mM과 1.22mM의 Km 값을 나타냈다. 그러나 금속이온의 효과에 대해 두 효소가 뚜렷한 차이를 보여 $Ba^{++}$$Mg^{++}$이 CM I의 활성은 약간 저해하는데 반해 CMII는 약 7% 활성화시켰다. 두 효소는 비교적 넓은 범위의 pH에서 안정하며 4$^{\circ}C$에서 10일간 저장했을 때 CM I이 75%, CMII 가 69%의 잔여 효소활성을 보였고 0.1mM EDTA 또는 0.1mM CoCl$_2$의 첨가는 CMII만을 약간 안정화시키는 효과를 보였고 CoCl$_2$는 CM I에 대해 오히려 불활성화 효과를 나타냈다. 열에 대해서는 불안정했으며 특히 CMII가 더욱 불안정하였다. CM I과 CMII는 pCMB에 의해 현저한 활성저해를 받았으며 저해정도는 두 효소가 큰 차이를 보여 catalytic site의 구조적 차이를 암시해주고 있다.

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

  • 허원녕
    • 한국식품과학회지
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    • 제27권5호
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    • pp.666-672
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    • 1995
  • Valencia orange로부터 분리된 TLPE는 분자량이 36 KD(37.5 KD)이며 등전점이 pH 8.4, 8.7, 8.9, 9.8, ${\geq}10$인 isoenzyme의 혼합물이었고, 또한 TSPE는 분자량이 53 KD(50 KD)이며 등전점이 pH 8.7, 9.2, ${\geq}10$인 isoenzyme의 혼합물이었다. TLPE의 최적 작용 pH는 $7.0,\;appK_{M}$은 1.1 mg/ml, appVmax는 $0.53\;{\mu}mol/min/{\mu}g$, turnover number는 19,000 mol/mol/min이었으며, TSPE의 최적 작용 pH는 $7.0{\sim}8.5,\;appK_{M}$은 1.7 mg/ml, appVmax는 $1.01\;{\mu}mol/min/{\mu}g$, turnover number는 54,000 mol/mol/min이었다. TSPE는 가열처리와 오렌지쥬스에서의 저장에 대하여 TLPE보다 훨씬 안정하였다. 또한 PE조효소, TSPE 그리고 TLPE 중에서는 orange juice의 cloud loss에 PE조효소가 가장 크게 영향을 미쳤으며 그 다음으로 TSPE TLPE 순서로 나타났다.

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Methylene Blue의 방사선 방어결과 1. Methylene Bluerk가 $\gamma$선에 조사된 흰쥐의 유산탈수소효소의 함량과유산탈수소효소의 동위효소에 미치는 영향 (Radioprotective Effect of Methylene Blue 1. Effect of Methylene Blue on Lactic Dehydrogenase Level and Lactic Dehydrogenase Isoenzymes of Rats Exposed to Gamma-Irradiation)

  • Sang Yul Nam;Seung Han Chang
    • 한국동물학회지
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    • 제12권3호
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    • pp.94-102
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    • 1969
  • 盛熱한 albino 雄性 흰쥐를 對照群과 實驗群으로 나누어 對照群에는 0.9% 生理食鹽水를, 實驗群에는 methylene blue(38mg/kg, 食鹽水에 溶解 pH 7.4)를 各各 腹腔注射處理하였다. 約 30 分後 兩群 共히 總線量 360 rads 인 $^60 Co$의 r 線을 一時 全身照射하였다. 照射後 212時間에 걸쳐 血淸總乳酸脫水素酵素의 含量(Cabaud and Droblewski 1958)과 血淸을 비롯하여 肝, 心臟, 精巢組織의 乳酸脫水素酵素의 同位酵素vnfortys(Preston et al., 1965)을 各各 測定하여 methylene blue 의 放射線에 對한 防禦 果를 硏究調査하였다. 一般的으로 血淸總乳酸脫水素酵素의 含量은 兩群共히 r 線照射後 初期에 增加하나 特히 methylene blue 實驗群인 處理群은 對照群인 生理食鹽水群에 比하여 15 및 64 時間區에서 增加의 遲滯性을 나타내었다. 血淸과 肝, 心臟 및 精巢組織의 乳酸脫水素酵素의 同位酵素는 兩群共히 線照射後 40과 116 時間區에서 陽極쪽의 同位酵素 푸랙숀 含量은 減少한다. 照射群 methylene blue 處理群의 血淸과 肝 및 精巢組織 乳酸脫水素酵素의 同位酵素패턴에서는 對照群과 別差異가 없었으나 特히 methylene blue 處理群의 心臟組織은 對照群에 比하여 若干의 增減의 遲滯性을 나타내었다. 兩群共히 照射後 血淸乳酸脫水素酵素含量의 增加는 同位酵素푸랙숀 1의 增加에 基因된다고 思料되며 이상의 結果로 미루어 보아 總乳酸脫水素酵素 活性과 心臟組織의 乳酸脫水素酵素의 同位酵素패턴에 있어서 methylene blue가 放射線防禦 果가 있는 것으로 思料된다.

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닭의 혈청 Creatine phosphokinase (CPK) 총 활성 및 CPK isoenzymes분획에 관한 연구 (Studies on the Serum Total Creatine Phosphokinase (CPK) Activities and CPK Isosenzymes Fractions of Chicken)

  • 김봉식;허인;김종형;장명환;김원선;김덕환
    • 한국동물위생학회지
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    • 제16권2호
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    • pp.120-126
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    • 1993
  • Clinically healthy 66 female Maniker breed chicken(22 of 2-week-old : group A, 23 of 8-week-old: group B and 21 of 27-week-old: group C) and 66 female Manina breed chicken (21 of 2-week-old : group D, 22 of 8-week-old : group E and 23 of 27-week-old : group F) were examined to establish physiological basic data on serum total Creatine phosphokinase (CPK) activities and CPK isoenzymes fractions. The results obtained were summarized as follows; 1. Serum total CPK activities were $1,088{\pm}254.0\;IU\;/{\ell},$ $1,454{\pm}337.2\;IU\;/{\ell}$ and $1,440{\pm}526.3\;IU\;/{\ell}$ in gorup A, group B and group C of Maniker breed, respectively. Group B and group C showed higher values than that of group A (P<0.01) meaning high values nth aging. 2. Serum total CPK activities were $1,676{\pm}420.5\;IU\;/ {\ell},$ $1,007{\pm}283.1\;IU\;/{\ell}$ and $862{\pm}294.5\;IU/{\ell}$ in group D, group E and group F of Manina breed, respectively. Group D showed the highest value among groups (P<0.01) and Manina breed showed the low values of serum total CPK activities with aging. 3. Manina breed at 2-week-old and Maniker breed at 8-week-old and 27-week-old showed significant high values of total serum CPK activities in breed differance (P<0.01) 4. In the pattern of serum CPK isoenzyme fractions, group A and group B were high with decreasing order of $CK_3>CK_2>CK_1$ and group C was high with decreasing order of $CK_2> CK_3>CK_1$ Groihp D, E and F showed the same pattern with decreasing order of $CK_2>CK_3> CK_1.$ 5. Significance of CPK isoenzymes fractions in breed differance was only found at 8-week-old. $CK_1\;and \;CK_3$ in Maniker breed (P<0.05), and $CK_2$ in Manina breed were higher than that of the other breed (P < 0.01).

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Mechanistic Insights into the Chemopreventive Action of Phenethy1 Isothiocyanate against Ν-Nitrosobis(2-Oxopropyl) Amine-Induced Carcinogenesis

  • Lee, In-Seon
    • Preventive Nutrition and Food Science
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    • 제1권2호
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    • pp.227-229
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    • 1996
  • The effects of phenthyl isothiocyanate(PEIFTC) on xenobiotic metabolizing enzymes and cell kinetics in the target organs for Ν-nirtosobis(2-oxopropyl) amine(BOP)-tumorigenicity were investigated in female Syrian golden hamsters in order to gain the mechanistic insigths into the chemopreventive action of PEITS against BOP-initiated lung and pancreatic carcinogenesis in hamsters. Hamsters were given BOP subcuteneo-usly(s.c.) and/or PEITC by gavage 2h prior to the BOP treatment. Eight and 24h after the PEITC administration, animals were sacrificed for analyzing P450 isoenzymes, glutathine(GSH), glutathione S-transferase(GST) and cell kinetics. The PEITC pretreatment significantly reduced the hepatic P450 isoenzume levels such as CYP2B1 and DYP1A1 which were significantly increased by the BOP treatment. However, PEITC did not affect the CYP levels in the pancreas and lung. Interestingly, the PEITC pretreatment rather lowered the heparic GST and GSH levels, regradless of BOP administration. Proliferating cell nuclear antigen(PCNA)- labeling indices were dose dependently decreased by PEITC in the pancreas acini and ducts, bronchioles, and renal tubules in which the cell replication was significantly affected by BOP. These results thus suggest that PEITC exerts the chemopreventive effects in hamsters by influencing xenobiotic matabolizing phase I enzymes in the liver and regulating cell kinetics in the target organs.

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