사과 과실로부터 분리한 polygalacturonase-inhibiting protein(PGIP)의 생화학적 특성 Characterization of an Apple Polygalacturonase-inhibiting Protein (PGIP) from Apple Fruits원문보기
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. Op...
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).
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).
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이론/모형
Initial rate kinetics were calculated using the Michaelis- Menton equation. To determine the effects of PGIP on the Vmax and Km values of PG from B.
Active fractions (2 mL) were pooled and concentrated as described above. Protein was measured by using Bradford method [2].
1). The effects of 'Fuji' PGIP on initial rate kinetics of B. do thidea PG were performed using a Lineweaver-Burk analy sis (Fig. 2). The Km and Vmax values of PG from B.
성능/효과
dothi dea successfully degraded cell wall components by measuringthe release of uronic acid from pectic cell wall material. These results demonstrated that PG from B. dothidea cleave the a-1, 4 linkages between D-galacturonic acid residues in the rhamnogalacturonan, the main component of pectin, and cause the separation of cell walls from each other and the maceration of host tissue.
참고문헌 (21)
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
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
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
Collmer, A. and N.T. Keen. 1986. The role of pectic enzymes in plant pathogenesis. Annu. Rev. Plant Dis. 24, 383-409
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
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
Favaron, F., R. D'Ovidio, E. Porceddu and P. Alghisi. 1994. Purification and molecular characterization of soybean polygalacturonase-inhibiting protein. Planta 195, 80-87
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
Gross, K. C. 1982. A rapid and sensitive method for assaying polygalacturonase using 2-cyanoacetamide. HortScience 17, 933-934
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
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
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
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)
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
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
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
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
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
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
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
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