• Title/Summary/Keyword: peroxidase isozyme

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

  • 김명원
    • Journal of Plant Biology
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    • v.29 no.4
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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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Purification and Characterization of Peroxidase Isozyme C from Mung Bean (녹두의 Peroxidase Isozyme C의 생화학적 성장)

  • Lee, Sang-Kap;Park, Woo-Churl
    • Applied Biological Chemistry
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    • v.30 no.3
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    • pp.219-226
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    • 1987
  • Peroxidase isozyme C was isolated from mung bean cotyledon and purified to homogeneity as ascertained by chromatography and polyacrylamide gel electrophoresis, and then crystallized. Purification procedures included ammonium sulfate precipitation and column chromatography on Sephadex G-75, DEAE-cellulose and DEAE-Sephadex A-50. Peroxidase isozyme C was purified about 63 fold with 5% recovery. Isozyme C showed optimal activity at pH 5.0 with o-dianisidine and at pH 6.0 with guaiacol as substrate, and the optimal temperature was $70^{\circ}C$. Molecular weight of 50,000 was estimated for the isozyme C by SDS-polyacrylamide gel electrophoresis. At $70^{\circ}C$, it took 30 min to inactivate the isozyme to 50%, and at $80^{\circ}C$, this isozyme was almost completely inactivated in 20 min. The Km value of isozyme C for o-dianisidine was 0.11mM and that for guaiacol was 60.98mM using hydrogen peroxide as cosubstrate, and the kinetic pattern showed a competitive cyanide inhibition with respect to substrate. The crystalline structure of isozyme C was rectangular in shape.

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Changes in Isozyme Patterns of Peroxidase and Esterase during the Microtuberization of Potato(Solanum tuberosum) (감자(Solanum tuberosum)의 기내 소괴경 형성 단계에 따른 Peroxidase와 Esterase 동위효소의 양상 변화)

  • 정현숙
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.51-57
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    • 1993
  • The microtuber was efficiently formed on SH medium containing 9% sucrose from the in vitro propagated shoot of potato (Solanum tuberosum cv. Sumi). In order to investigate gene expression depending on the development stage of microtuber, we examined the changes of peroxidase and esterase activities, and their isozyme patterns as well. Peroxidase and esterase activities were the highest at the 7 day-culture of the microtuber and subsequently decreased on the stage of microtuberization, whereas esterase activity increased at the stage of 60 day-culture. However, their activities in the ordinary tuber were higher than those of 60 day-cultured microtuber. In addition, in the peroxidase isozyme pattern two new bands of pI 7.05 and pI 4.65 were appeared at the 15- day and 60 day-cultures, respectively, as shown by isoelectric focusing. Various bands in the sterase isozyme pattern were shown at the 7 day-culture, and the band patterns were a large difference, comparing those of shoot and tuber. New bands in the esterase isozyme pattern also appeared at the 15 day- (pI4.52) and 60 day-cultures (pI 4.48). These results suggest that the changes of peroxidase and esterase activities and isozyme patterns are an important factor in the differentiation and development of potato.

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Isozyme Patterns on Esterase, Peroxidase and Amylase in Benincasa hipspida (Thunb) Cogn and Commelina communis L. (동아와 달개비의 전기영동에 의한 Isozyme의 변이)

  • 채규인;최정식;김영두
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.36 no.2
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    • pp.174-176
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    • 1991
  • A electrophoretic study on seveal important enzymes was conducted with seed, leaf and stem of Benincasa hipida (Thunb) Cogn and Commelina communis L. Isozyme patterns of esterase were different from the parts and fruit of wax gourd appeared only one band. In the peroxidase isozyme, fruit of wax gourd appeared four bands, but stem of common dayflower no appeared. In the amylase isozyme, seed and fruit of 'gourd were appeared clearly one band, respectively.

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A New Strategy for Determining Optimum pH of Isozymes

  • Yoon, Kil-Joong
    • Bulletin of the Korean Chemical Society
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    • v.25 no.7
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    • pp.997-1002
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    • 2004
  • A hydrogenperoxide sensor containing peroxidase extracted from horseradish was constructed and pH effect on its sensing ability was investigated. Current profiles of the biosensor with pH and the electrophoretic analysis showed that horseradish peroxidase consists of two isozymes. Assuming that it is a hypothetical twoisozyme mixture, the current profiles were deconvoluted into two Gaussians. Application of the new Michaelis-Menten equation connoting pH concept to this system enabled to find all the related dissociation constants of the isozyme-substrates and the isozyme-proton complexes and to determine pHs for the maximal isozyme activities.

Changes in Isozyme Patterns of Peroxidase and Esterase during Regeneration of Pepper (Capsicum annuum L.) (고추 (Capsicum annuum L.)의 재분화에 따른 Peroxidase와 Esterase 동위효소의 양상 변화)

  • 정현숙
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.9-15
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    • 1992
  • The plantlet was regenerated on MS medium containing BAP (2 mg/I) and 1M (1 mg/I) from leaf discs of pepper after 3 weeks of culture. And then, we investigated the activity of peroxidase and esterase and the pattern of their isozymes from leaf, stem and root in order to observe physiological and biochemical changes on the developemental stage, respectively. The peroxidase was expressed with tissue specificity because peroxidase activity according to the developemental stage of the tissue was not only highest in the leaf of the pepper at 10 days after it germinated but also 2 new bands of its isozyme were found in pI 7.2 and pI 5.2. However, a new pI 3.4 band was found in the leaf and root of the pepper after 14 days of germination, and in the stem was found out pI 5.2 band. As regeneration of leaf dises was progressed, its peroxiase activity was increased about 80% more than that of control after 14 days of culture and new pI 3.2 and 6.5 bands of it isozyme were found. The results suggested that peroxidase would be connected with regeneration of pepper. Also, esterase activity was increased about 50% more than that of control after 14 days of culture, the pattern of esterase isozyme was shown to be 3 cathodic bands and 1 anodic band after 7 days of culture.ulture.

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Studies on the Isozyme Patterns Followed by Redifferentiation Stages of the Pepper(Capsicum annuum L.) - Changes of isozyme pattern on somatic embryogenesis in cutured pepper cells - (고추(Capsicum annuum L.)의 재분화 단계에 따른 Isozyme 양상에 관한 연구 - 고추 배양세포의 체세포배 형성에 따른 Isozyme 양상의 변화-)

  • 정현숙;공현진박진수강영희
    • KSBB Journal
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    • v.7 no.3
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    • pp.216-221
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    • 1992
  • The physiological changes of somatic embryogenesis in cultured pepper cells (Capsicum annuum L. cv Shinhong) were investigated. The somatic embryogenesis was induced by cultivating the callus in hormone-free MS medium. The peroxidase patterns in the somatic embryogenic cells and the culture medium was revealed three and two of cathodic and anodic bands by isoelectric focusing respectively. Activity of peroxidase released into culture medium was 4 times higher than that of 12th day cultured cells. At the heart stage, the isozyme patterns of the MDH and esterase were found to be changed, which were showed by starch gel electrophoresis. It means these isozymes can be used as markers for studying somatic embryogenesis and differentiation.

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Amino Acid Sequence Studies of Basic Isozyme of Horseradish Peroxidase (서양고추냉이 Peroxidase의 염기성 Isozyme의 아미노산 배열에 관한 연구)

  • 이진영;방병호
    • The Korean Journal of Food And Nutrition
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    • v.8 no.1
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    • pp.37-42
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    • 1995
  • The amino acid sequence of basic isozyme 55 of Horseradish Peroxidase (HRP E5) was determined by protein sequencing. HRP E5 consisted about 300 residues, and has a molecular weight of approximately 36,000 $\pm$ 500 dalton. The protein was rich In aspartic acid (14%), arginine(13%), and leucine(11%). The primary structure of HRP E5 was established by sequencing its tryptic (T1-T19) and lysylendopeptic (Al-A3) peptides. The sequence homology between HRP E5 and HRP C (neutral isozyme of horseradish peroxidase) is found to be more than 66%. The highest concentration of identical residues are found on residues 29~56, 90~123, and 155~173, but relatively low on 174~271.

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Electrophoretic Patterns of Isozymes from the Mycelia of the Auxotrophs of Lentinula edodes (표고버섯 영양요구성 변이주의 전기영동법에 의한 Isozyme 비교)

  • Kim, Chae-Kyun;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.25 no.2 s.81
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    • pp.85-90
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    • 1997
  • The Isozyme activities of Lentinula edodes were studied as a preliminary study for genetic analysis after protoplast fusion. The presence of peroxidase, esterase, superoxide dismutase, acid phosphatase, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ was examined. An intracellular buffer-soluble protein from the mycelia was used for enzyme analysis on nondenaturing polyacrylamide gels. The auxotrophs of Lentinula edodes were positive for peroxidase, esterase, superoxide dismutase and acid phosphatase. However, alkaline phosphatase, alcohol dehydrogenase and ${\alpha}-amylase$ were not detected. The esterase and peroxidase were not affected by the various culture age. Isozyme identification may be a useful tool after protoplast fusion.

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