• Title/Summary/Keyword: zymogram

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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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Studies on Esterase of Pieris rapae L. I. Changes of Esterase Activity and Zymogram Pattern During Development and Purification (배추흰나비(Pieris rapae L.)의 esterase에 관한 연구 I. 변태에 따른 esterase의 활성변화 및 zymogram pattern의 변화와 정제)

  • 박철호;김학열;여성문
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.330-336
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    • 1990
  • Changes in esterase activity and zymogram pattern during development in Pieirs rapae L. were investigated and three esterases (E2, E6 and E11) from the late 5th instar larvae were purified. Esterase activity in whole body increased rapidly during 5th instar larval stages and reached a peak at the late 5th instar larval stage. The number and intensity of esterase band from whole body and midgut also showed a peak at the late 5th instar larval stage. Purification of esterase was performed using gel filtration on Sephadex G-lOO, ion-exchange chromatography on DEAE-trisacryl and preparative electrophoresis. The final purities of these enzymes were about 30 to 60-fold.

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Responses of VA mycorrhizal Fungus, Glomus mosseae, on the Growth and Nutrition of Mulberry tree (VA 내생균근균, Glomus mosseae,가 뽕나무의 생장과 영양에 미치는 영향)

  • 김중채;문재곡
    • Journal of Sericultural and Entomological Science
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    • v.28 no.1
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    • pp.1-14
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    • 1986
  • This study was carried out to acguire some basic information on nutritional and lhysiological effects of vesicular arbuscular mycorrhizae(VAM) on mulberry trees inoculated with Glomus mosseae, Gerd. & Trap. grown in clay for 65 days and treated with 5 different levels of phosphorus, ie 30, 60, 120, 240, 480 ppm as (NH4)2 HPO4. At the End of the expermental period the levels of fixed phosphate in the soil was measared. And the native VAM fungi were collected to select the most effective VAM species on mulberry tree. The nutritional and biochemical effects of VA mycorrhizae on the mulberry leaves were also studied. Those results are as follow. 1. The mulberry trees grown in clay and inoculated with VAM were heavier in shoot dry weight as much as 197% than uninoculated plants. But in vermioulite, uninoculated mulberry trees were heavier as much as 135% than inoculated. 2. The rates of endo mycorrhizal formation in clay was highest at 60 ppm level of phosphorus, and vesicles in roots were formed in 240ppm and 480ppm level of phosphorus, but not in 30ppm, 60ppm and 120ppm. 3. The greatest growth responses of VAM inoculation was found at 60ppm level of hosphorus, and the optimum phosphorus level for VAM responese appeared to be 60ppm. 4. VAM was also to absorb soil-fixed phosphate. VAM abosrbed Fe-bound phosphate most efficiently and Ca-bound phosphate with ease but not Al-bound phosphate and Al-Fe occluded phosphate. 5. Three species of Gigaspora and one species of Glamus growing naturally in mulberry plantations were collected and tested for the growth responses. Gigaspora tricalipta and Gigaspora calospora revealed the greatest growth responses on mulberry tree among tested VAM fungi. 6. Mulberry leaves inoculated with VA mycorrhizal fungi contained 9.8% more phosphate and 15.2% more nitrogen, 22.2% more water-soluble carbohydrates and 15.2% more proteins than uninoculated plants. 7. The electrophoretic pattern of mulberry leaf protein inoculated with VAM fungi has 19 bands. 8. The patterns of peroxidase zymogram and Amulase zymogram were different between the mulberry leaves inoculated and uninoculated with VA mycorrhizal fungi. The peroxidase zymogram of inoculated leaves has 1 less majour band than unioculated leaves The amylase zymogram of inoculated leaves has 2 bands near the +pole, but that of uninoculated leaves has 1 band near the $pole.

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Comparison of Electrophoretic Isozyme Band Pattern of Pleurotus spp. in Korea -ll. Isoelectric Focusing- (한국산 느타리 버섯 (Pleurotus spp.)의 전기영동 Isozyme Band Pattern 비교 -II. 등전점 전기영동-)

  • Park, Yong-Hwan;Byun, Myung-Ok;Hiroshi, Fujii
    • The Korean Journal of Mycology
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    • v.16 no.2
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    • pp.95-100
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    • 1988
  • Isozyme band patterns were investigated by isoeletric focusing Esterase and Leucine amino peptidase of Pleurotus spp. in Korea. Esterase patterns of mycelia and fruitbody were distinquished. However, those of primordia, cap and stem were similar. Interspecies differences of Pleurotus ostreatus, P. cornucopiae and P. florida of Esterase zymogram were found. Species identification by electrophoretic zymogram may be a role as an additional taxonomic tool.

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Shattering-related Characteristics and Germination Speed Based on Grain Type and Esterase Isozyme Zymogram in Korean Off-rype Rice

  • Kim, Dong-Kwan;Jin, Il-Doo;Kim, Yeon-Gyu;Min, Kyung-Soo;Jung, Byung-Gwan;Kim, Yong-Jae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.1
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    • pp.29-35
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    • 2002
  • This study was carried out to investigate the characteristics of shattering habit and germination of offtype rice plants collected in Korea which were classified into seven groups based on grain characteristics. In the long-grain red rice group, the short-grain red rice group, the long-grain normal rice group, and the long-grain waxy rice group, the tensile strength of grain and primary rachis branch was relatively low and the fiber cell of the primary rachis branch was short. Characteristics of shattering-related traits and germination in relation to grain and esterase isozyme zymogram type of off-type rice plants. In most lines of the extremely late and sterile rice group, the tensile strength of grain was low, while that of primary rachis branch was high, and the fiber cell was long. However, in the type 1 esterase isozyme zymogram(EIZ) lines among the long-grain normal rice group and the type 6 and 5+6 EIZ plants among the extremely late and sterile rice group, the tensile strength of grain was relatively higher. The long-grain waxy rice group and the type 3 EIZ lines among the long-grain red rice group, showed higher germinability than did cultivars which are known to germinate well even in low-temperature. The other off-type rice group, except for the short-grain waxy rice group and type 1 and 7 EIZ lines among the long-grain normal rice group, had higher germination speed than that of the cultivars.

Comparison of the Influence of Carbon Substrates on the Fibrolytic Activities of Neocallimastix sp. NLRI-3 (탄소원의 종류가 반추위 혐기 곰팡이 Neocallimastix sp. NLRI-3의 섬유소 분해효소 활력에 미치는 영향 비교)

  • 손호진;송재용;최낙진;하종규;장종수
    • Journal of Animal Science and Technology
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    • v.48 no.3
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    • pp.415-424
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    • 2006
  • The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.

Characterization and Production of Thermostable and Acid-stable Extracellular Fibrinolytic Enzymes from Cordyceps militaris

  • Kim, Seon-Ah;Son, Hong-Joo;Kim, Keun-Ki;Park, Hyun-Chul;Lee, Sang-Mong;Cho, Byung-Wook;Kim, Yong-Gyun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.22 no.2
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    • pp.83-93
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    • 2011
  • Biochemical and enzymatic characterization for extracellular protease isolated from Cordyceps militaris cultivated on rice bran medium was investigated. C militaris produced proteolytic enzymes from 10 days after inoculation, maximum enzyme production was found at 25 days. The optimum temperature and pH of proteases production was at $25^{\circ}C$ and pH 7.0, respectively. The protease activity was observed in the four peaks (Pro-I, Pro-II, Pro-III, and Pro-IV) separated through Sephadex G-100 column chromatography. The separated protease was optimally active at $25^{\circ}C$. Optimum pH of the protease was between 7 and 8. Enzyme was also stable over at $30-80^{\circ}C$. The enzyme was highly stable in a pH range of 4-9. Protease activity was found to be slightly decreased by the addition of $Mg^{2+}$, $Mn^{2+}$, $Zn^{2+}$, $Fe^{2+}$ and $Cu^{2+}$, whereas inhibited by the addition of $Ca^{2+}$ and $Co^{2+}$ Protease activity was inhibited by protease inhibitor PMSF. On the other hand, the partially purified protease was investigated on proteolytic protease activity by zymogram gel electrophoresis using three substances (casein, gelatin and fibrin). Four active bands (F-I, FII, F-III, and F-IV) of fibrin degradation were revealed on fibrin zymogram gels. Both of F-II and FIII showed caseinolytic, fibrinolytic and gelatinolytic activities in three gels. Thermostability, pH stability, and pH-thermostability of the enzyme determined the residual fibrinolytic activity also displayed on fibrin zymogram gel. The only one enzyme (F-II) displayed over a broad range of temperature at $30-90^{\circ}C$. The FII displayed fibrinolytic activity in the pH range 3-5, but was inactivated in the range of pH 6-11. The F-I and F-III showed enzyme activity in the pH range of 6-11. In the pH-thermostability, the F-II only kept fibrinolytic activity after heating at $100^{\circ}C$ for 10, 20 and 30 min at pH 3 and pH 7, respectively. On the other hand, the F-II was retained activity until heating for 10 min under pH 11 condition. By using fibrin zymogram gel electrophoresis, extracellular fibrinolytic enzyme F-II from C. militaris showed unusual thermostable under acid and neutral conditions.

Isolation and Characterization of Thermostable Xylanase-producing Paenibacillus sp. DG-22. (내열성 Xylanase를 생산하는 Paenibacillus sp. DG-22 균주의 분리 및 효소 특성)

    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.22-28
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    • 2004
  • A new moderate thermophilic bacterial strain DG-22 which produces thermostable xylanase was isolated from a timber yard soil in Kyungju, Korea. On the basis of morphological, biochemical and phylogenetic studies the new isolate was identified as a Paenibacillus species. Production of xylanase in this strain was strongly induced by adding xylan to the growth medium and repressed by glucose or xylose. No cellulase activity was detected. The temperature and pH for optimum activity were 8$0^{\circ}C$ and 5.0-5.5, respectively. The crude xylanase was stable at $60^{\circ}C$ and retained 60% of initial activity after 2h at $70^{\circ}C$. Zymogram analysis of the culture supernatant showed two xylanase active bands with molecular masses of 22 and 30 kDa.

Comparison of Three Substrates (Casein, Fibrin, and Gelatin) in Zymographic Gel

  • Choi, Nack-Shick;Yoon, Kab-Seog;Lee, Jin-Young;Han, Kyoung-Yoen;Kim, Seung-Ho
    • BMB Reports
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    • v.34 no.6
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    • pp.531-536
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    • 2001
  • Three zymographic techniques using casein, fibrin, and gelatin as substrates in SDS-PAGE were compared based on three aspects: (1) The proteolytic pattern of extracellular enzymes from the three bacterial strains, Bacillus sp. DJ-1, DJ-2, and DJ-3. (2) The enzymatic sensitivity of their activity on zymogram gels. (3) The stability of stained zymogram gels with Coomassie brilliant blue in the destaining solution. There was no significant difference on the pattern of extracellular enzymes from the three strains. The bands in the fibrin gel were clearer and more distinct from the extensive destaining process. It was also shown that the gelatin gel revealed the highest enzymatic sensitivity among the three gels, based on the densitometric analysis. In the casein gel, a trace that could be mistaken as a proteolytic band appeared around 40-50 kDa.

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