생명과학회지 (Journal of Life Science)

  • 제16권4호
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  • Pages.653-658
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  • 2006
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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사과 과실로부터 분리한 polygalacturonase-inhibiting protein(PGIP)의 생화학적 특성

Characterization of an Apple Polygalacturonase-inhibiting Protein (PGIP) from Apple Fruits

  • 이동훈 (농촌진흥청 난지농업연구소 감귤과) ;
  • 강상구 (영남대학교 생명공학부) ;
  • 강인규 (상주대학교 환경원예학과) ;
  • 이윤경 (아시아대학교 한약자원학과) ;
  • 최철 (경북대학교 식물생명과학부) ;
  • 변재균 (영남대학교 자연자원학부)
  • Lee, Dong-Hoon (Citrus Research Division, National Institute of Subtropical Agriculture, RDA.) ;
  • Kang, Sang-Gu (School of Biotechnology, Institute of Biotechnology, Yeungnam Univ.) ;
  • Kang, In-Kyu (Dept. of Enviromental Horticulture, Sangju National Univ.) ;
  • Lee, Yoon-Kyeong (Dept. of Oriental Medicine Resources, Asia Univ.) ;
  • Choi, Cheol (Division of Plant Biosciences, Kyungpook National Univ.) ;
  • Byun, Jae-Kyun (School of Natural Resources, Yeungnam Univ.)
  • 발행 : 2006.07.31
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초록

사과 겹무늬썩음병균이(Botryosphaeria dothidea) 생성하는 세포벽 분해효소인 endopolygalaturonase를 억제하는 polygalacturonase-inhibiting protein (PGIP)를 사과 과실로부터 분리하였다. 분리되어진 사과 PGIP는 사과 겹무늬썩음병균이 생성하는 PG에 대하여 혼합형의 저해를 나타내었다. PGIP의 반응 최적온도는 $40^{\circ}C$이며 최적 pH는 5.0이었다. 이 효소는 $60^{\circ}C$까지는 비교적 안정하였으나 $70^{\circ}C$에서는 효소의 활성이 완전히 억제되었으며 pH 4.0에서 8.0까지는 안정하였다. PGIP는 $K^+$, $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$ 과 $Zn^{2+}$ 등의 금속이온과 SDS 그리고 CDTA에 의해 효소의 활성이 저해되었다.

An apple polygalacturonase-inhibiting protein (PGIP), that specifically inhibited endopolygalacturonase (PG, EC 3.2.1.15) from Botryosphaeria dothidea, was purified from B. dothidea infected apple (Malus domestica cv. Fuji) fruits. The apple PGIP was a mixed-type inhibitor of PG from B. dothidea. Optimal temperature for the maximum enzyme activity was $40^{\circ}C$, and optimum pH of the purified PGIP was pH 5.0. PGIP was stable up to temperature of $60^{\circ}C$ and was completely suppressed after heating at $70^{\circ}C$ for 10 min, PGIP was stable at pH between 4 and 8. Inhibition of PG by PGIP was reduced by $K^+$, $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$ metal ion, sodium dodecyl sulfate (SDS) and 1,2-diaminocyclohexane tetra acetate (CDTA).

키워드

참고문헌

  1. Abu-Goukh, A. A., L. C. Greve and J. M. Labavitch. 1983. Purification and partial characterization of 'Barlett' pear fruit polygalacturonase inhibitors. Physiol. Plant Pathol. 23, 111-122 https://doi.org/10.1016/0048-4059(83)90038-3
  2. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  3. Cervone, F., G. De Lorenzo, L. Degra, G. Salvi and M. Bergami. 1987. Purification characterization of a polygalacturonase- inhibiting protein from Phaseolus vulgaris L. Plant Physiol. 85, 631-637 https://doi.org/10.1104/pp.85.3.631
  4. Collmer, A. and N.T. Keen. 1986. The role of pectic enzymes in plant pathogenesis. Annu. Rev. Plant Dis. 24, 383-409
  5. Conway, W. S., K. C. Gross, C. D. Boyer and C. E. Sams. 1988. Inhibition of Penicillium expansum polygalacturonase activity by increase apple cell wall calcium. Phytopathology 78, 1052-1055 https://doi.org/10.1094/Phyto-78-1052
  6. Cooper, R. M. 1984. The role of cell wall-degrading enzymes in infection and damage. in : Plant Diseases: Infection, Damage and Loss. R.K.S. Wood and G.J.Jellis, eds. Blackwell Scientific Publications, Oxford, p. 13-27
  7. Favaron, F., R. D'Ovidio, E. Porceddu and P. Alghisi. 1994. Purification and molecular characterization of soybean polygalacturonase-inhibiting protein. Planta 195, 80-87
  8. Favaron, F., C. Castiglioni, R. D'Ovidio and P. Alghisi. 1997. Polygalacturonase inhibiting proteins from Allium porrum L. and their role in plant tissue against fungal endo- polygalacturonases. Physiol. Mol. Plant Pathol. 50, 403-417 https://doi.org/10.1006/pmpp.1997.0099
  9. Gross, K. C. 1982. A rapid and sensitive method for assaying polygalacturonase using 2-cyanoacetamide. HortScience 17, 933-934
  10. James, J. T. and I. A. Dubery. 2001. Inhibition of polygalacturonase from Verticillium dahliae by a polygalacturonase inhibiting protein from cotton. Phytochemistry 57, 149-156 https://doi.org/10.1016/S0031-9422(01)00024-3
  11. Johnston, D. J. and B. Williamson. 1992. Purification and characterization of four polygalacturonase from Botrytis cinerea. Mycol. Res. 96, 343-349 https://doi.org/10.1016/S0953-7562(09)80948-9
  12. Johnston, D. J., V. Ramanathan and B. Williamson. 1993. A protein from immature raspberry fruits which inhibitis endopolygalacturonases from Botrytis cinerea and other micro- organisms. J. Exp. Bot. 44, 971-976 https://doi.org/10.1093/jxb/44.5.971
  13. Kemp, G., C. W. Bergmann, R. Clay, A. J. Van der Westhuizen and Z. A. Pretorius. 2003. Isolation of a polygalacturonase- inhibiting protein(PGIP) from wheat. Mol. Plant-Microbe Interact. 16, 955-961 https://doi.org/10.1094/MPMI.2003.16.11.955
  14. Lee, D. H., H. Bae, I. K. Kang, J. K. Byun and S. G. Kang. 2006. Characterization of an Apple Polygalacturonase-inhibiting Protein(PGIP) That Specifically Inhibits an Endopolygalacturonase(PG) Purified from Apple Fruits Infected with Botryosphaeria dothidea. J. Microbiol. Biotechnol. (in press)
  15. Manachini, P. L., M. G. Fortina and C. Parini. 1987. Purification and properties of an endopolygalacturonase produced by Rhizopus stolonifer. Biotechnol. Lett. 9, 219-224 https://doi.org/10.1007/BF01024570
  16. Park, S. H., S. G. Suh and C. U. Lee. 1997. Purification and N-termina amino acid sequence of polygalacturonase produced by Botryosphaeria dothidea. Korean J. Plant Pathol. 13, 402-407
  17. Peretto, R., F. Favaron, V. Bettini, G. De Lorenzo, S. Marini, P. Alghisi, F. Corvone and P. Bonfante. 1992. Expression and localization of polygalacturonase during the outgrowth of lateral roots in Allium porrum L. Planta 188, 164-172 https://doi.org/10.1007/BF00216810
  18. Stotz, H. U., A. L. T. Powell, S. E. Damon, L. C. Greve, A. B. Bennett and J. M. Labavitch. 1993. Molecular characterization of polygalacturonase inhibitor from Pyrus communis L. cv. Barlett. Plant Physiol. 102, 133-138 https://doi.org/10.1104/pp.102.1.133
  19. Toubart, P., A. Desideri, G. Salvi, F. Cervone, L. Daroda, G. De Lorenzo, C. Bergmann, A. G. Darvill and P. Albersheim. 1992. Cloning and characterization of the endo- polygalacturonase-inhibiting protein(PGIP) of Phaseolus vulgaris L. Plant J. 2, 367-373
  20. Van der Cruyssen, G., E. De Meester and O. Kamoen. 1994. Expression of polygalacturonase of Botrytis cinereain in vitro and in vivo. Meded. Fac. Landbouwk. Toegep. Biolog. Wetensch. Univ. Gent. 59, 895-905
  21. Yao, C., W. S. Conway and C. E. Sams. 1995. Purification and characterization of a polygalacturonase-inhibiting protein from apple fruit. Phytopathology 85, 1373-1377 https://doi.org/10.1094/Phyto-85-1373