Hostname: page-component-cd9895bd7-jn8rn Total loading time: 0 Render date: 2024-12-25T15:50:07.087Z Has data issue: false hasContentIssue false
  • English
  • Français

Soybean seed pectinesterase

Published online by Cambridge University Press:  19 September 2008

Oskar Markovič  and
Ralph L. Obendorf
Oskar Markovič
Affiliation:
Seed Biology, Department of Soil, Crop and Atmospheric Sciences, Cornell University Agricultural Experiment Station, 619 Bradfield Hall, Cornell University, Ithaca, NY 14853-1901, USA Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK 842 38 Bratislava, Slovakia
Ralph L. Obendorf*
Affiliation:
Seed Biology, Department of Soil, Crop and Atmospheric Sciences, Cornell University Agricultural Experiment Station, 619 Bradfield Hall, Cornell University, Ithaca, NY 14853-1901, USA
*
*Fax: +1 607 255 2644 E-mail: rlol@cornell.edu
Get access Rights & Permissions [Opens in a new window]

Abstract

Methanol accumulates in axis tissues of maturing soybean seeds, correlating with preharvest seed deterioration. Accumulation of methanol appears to be associated with the enzymic demethylation of pectin methyl esters by pectinesterase (PE; EC 3.1.1.11). To characterize PE in developing and maturing soybean (Glycine max (L.) Merrill) seeds, enzyme activity was assayed in axis and cotyledon tissues. Activity per g fresh weight was 20–25 times higher in axes than in cotyledons with highest activities between 45 and 60 days after flowering (DAF). Twenty to 33% of the total PE activity was in the ‘soluble’ form (extracted with water, 0.5 M sucrose, 1 M sucrose and water). Soluble and cell-wall-bound PE (subsequently extracted with 1 M NaCI) were purified and characterized from axes of seeds at 45–60 DAF. Purification of PE was achieved through concentration of extracts by ultra-filtration, precipitation with ammonium sulfate (30–80% saturation), dialysis, gel filtration on Sephadex G-75 columns, and ion exchange chromatography on CM Sepharose CL-6B. Further purification of both soluble and bound PE was by isoelectric focusing (IEF) on ultrathin layers of polyacrylamide gel with simultaneous detection of protein and PE activity. It was possible to follow seven bands exhibiting PE activity with pl values between 6.0 and 9.5 in 1 M NaCI-extracts of total homogenates. Differences in the IEF patterns of bound and soluble PE were observed. Whereas the bound enzyme exhibited more basic PE bands (pl 8–9.5), the soluble enzyme had more active bands at pl 6.5, 7.0 and 7.5. The Mr was close to 33 000 and the pH optimum was 7.8 for both soluble and bound PE.

Keywords

cell wallenzyme activityisoelectric focusing (IEF)pectinesterase (EC 3.1.1.11)seed developmentsoybean (Glycine max (L.) Merr.)
Type
Physiology & Biochemistry
Information
Seed Science Research , Volume 8 , Issue 4 , December 1998 , pp. 455 - 461
Copyright
Copyright © Cambridge University Press 1998

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Alonso, J., Rodríguez, T. and Canet, W. (1995) Effect of calcium pretreatments on the texture of frozen cherries. Role of pectinesterase in the changes in the pectic materials. Journal of Agricultural and Food Chemistry 43, 1011–1016.CrossRefGoogle Scholar
Bordenave, M. and Goldberg, R. (1993) Purification and characterization of pectin methylesterases from mung bean hypocotyl cell walls. Phytochemistry 33, 999–1003.CrossRefGoogle Scholar
Bordenave, M. and Goldberg, R. (1994) Immobilized and free apoplastic pectinmethylesterases in mung bean hypocotyl. Plant Physiology 106, 1151–1156.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
Charnay, D., Nari, J. and Noat, G. (1992) Regulation of plant cell-wall methyl esterase by polyamines – Interactions with the effects of metal ions. European Journal of Biochemistry 205, 711–714.CrossRefGoogle ScholarPubMed
Delincée, H. (1976) Thin-layer isoelectric focusing of multiple forms of tomato pectinesterase. Phytochemistry 15, 903–906.CrossRefGoogle Scholar
Frenkel, C., Peters, J.S., Tieman, D.M., Tiznado, M.E. and Handa, A.K. (1998) Pectin methylesterase regulates methanol and ethanol accumulation in ripening tomato (Lycopersicon esculentum) fruit. Journal of Biological Chemistry 273, 4293–4295.CrossRefGoogle ScholarPubMed
Goldberg, R. (1984) Changes in the properties of cell wall pectin methylesterase along the Vigna radiata hypocotyl. Physiologia Plantarum 61, 58–63.CrossRefGoogle Scholar
Goldberg, R., Pierron, M., Durand, L. and Mutaftschiev, S. (1992) In vitro and in situ properties of cell wall pectin methylesterases from mung bean hypocotyls. Journal of Experimental Botany 43, 41–46.CrossRefGoogle Scholar
Hagerman, A.E. and Austin, P.J. (1986) Continuous spectrophotometric assay for plant pectin methyl esterase. Journal of Agricultural and Food Chemistry 34, 440–444.CrossRefGoogle Scholar
Honig, D.H. and Rackis, J.J. (1975) Volatile components of maturing soybeans. Cereal Chemistry 52, 396–402.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage phage T4. Nature 227, 680–685.CrossRefGoogle Scholar
Markovič, O., Slezárik, A. and Labudová, I. (1985) Purification and characterization of pectinesterase and polygalacturonase from Trichoderma reesei. FEMS Microbiology Letters 27, 267–271.CrossRefGoogle Scholar
Markovič, O. and Obendorf, R.L. (1994) Soybean seed pectinesterase. Bratislava Symposium on Saccharides: Program and Abstracts 7, 109.Google Scholar
Moustacas, A.M., Nari, J., Borel, M., Noat, G. and Ricard, J. (1991) Pectin methylesterase, metal ions and plant cell-wall extension. The role of metal ions in plant cell-wall extension. Biochemical Journal 279, 351–354.CrossRefGoogle ScholarPubMed
Moustacas, A.M., Nari, J., Diamantidis, G., Noat, G., Crasnier, M., Borel, M. and Ricard, J. (1986) Electrostatic effects and the dynamics of enzyme reactions at the surface of plant cells. 2. The role of pectin methyl esterase in the modulation of electrostatic effects in soybean cell walls. European Journal of Biochemistry 155, 191–197.CrossRefGoogle ScholarPubMed
Nari, J., Noat, G., Diamantidis, G., Woudstra, M. and Ricard, J. (1986) Electrostatic effects and the dynamics of enzyme reactions at the surface of plant cells. 3. Interplay between limited cell-wall autolysis, pectin methyl esterase activity and electrostatic effects in soybean cell walls. European Journal of Biochemistry 155, 199–202.CrossRefGoogle ScholarPubMed
Nari, J., Noat, G. and Ricard, J. (1991) Pectin methylesterase, metal ions and plant cell-wall extension. Hydrolysis of pectin by plant cell-wall pectin methylesterase. Biochemical Journal 279, 343–350.CrossRefGoogle ScholarPubMed
Nemecek-Marshall, M., MacDonald, R.C., Franzen, J.J., Wojciechowski, C.L. and Fall, R. (1995) Methanol emission from leaves. Enzymic detection of gas-phase methanol and relation of methanol fluxes to stomatal conductance and leaf development. Plant Physiology 108, 1359–1368.CrossRefGoogle Scholar
Obendorf, R.L., Rytko, G.T. and Byrne, M.C. (1983) Soya bean seed growth and maturation by in vitro pod culture. Annals of Botany 51, 217–227.CrossRefGoogle Scholar
Obendorf, R.L., Koch, J.L., Górecki, R.J., Amable, R.A. and Aveni, M.T. (1990) Methanol accumulation in maturing seeds. Journal of Experimental Botany 41, 489–495.CrossRefGoogle Scholar
Ricard, J., Noat, G., Crasnier, M. and Job, D. (1981) Ionic control of immobilized enzymes. Kinetics of acid phosphatase bound to plant cell walls. Biochemical Journal 195, 357–367.CrossRefGoogle ScholarPubMed
Warrilow, A.G.S., Turner, R.J. and Jones, M.G. (1994) A novel form of pectinesterase in tomato. Phytochemistry 35, 863–868.CrossRefGoogle ScholarPubMed
Zhang, M., Nagata, S., Miyazawa, K., Kikuchi, H. and Esashi, Y. (1997) A competitive enzyme-linked immunosorbent assay to quantify acetaldehyde-protein adducts that accumulate in dry seeds during aging. Plant Physiology 113, 397–402.CrossRefGoogle ScholarPubMed
Zhang, M., Nakamaru, Y., Tsuda, S., Nagashima, T. and Esashi, Y. (1995a) Enzymatic conversion of volatile metabolites in dry seeds during storage. Plant and Cell Physiology 36, 157–164.Google Scholar
Zhang, M., Yoshiyama, M., Nagashima, T., Nakagawa, Y., Yoshioka, T. and Esashi, Y. (1995b) Aging of soybean seeds in relation to metabolism at different relative humidities. Plant and Cell Physiology 36, 1189–1195.Google Scholar