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Abstrakty
Uridine 5 '-diphosphoglucose-dependent glucosyltransferase which catalyzes the glucosylation of solasodine, i.e. UDP-glucose:solasodine glucosyltransferase, is present in leaves, roots, unripe fruits and unripe seeds of eggplant (Solanum melongenaL.). The glucosylation product is chromatographically identical with authentic solasodine 3β-nmonoglucoside, a putative intermediate in the biosynthesis of solasodine-based glycoalkaloids characteristic of the eggplant. The enzyme was purified about 50-fold from crude cytosol fraction of eggplant leaves by ammonium sulphate precipitation and column chromatography on Q-Sepharose and Sephadex G-100. The native enzyme has a molecular mass of approx. 55 and pH optimum of 8.5. metal ions are not required for its activity but the presence of free -SH groups is essential. Besides solasodine (Km = 0.04 ), the enzyme effectively glucosylates tomatidine, another steroidal alkaloid of the spirosolane type, but it is virtually inactive towards the solanidane-type steroidal alkaloids such as solanidine or demissid- ine. The enzyme is specific for UDP-glucose (Km= 2.1 u) since unlabelled ADP-, GDP-, CDP- or TDP-glucose could not effectively compete with UDP-[14]glucose used as the sugar donor for solasodine glucosylation. Moreover, no synthesis of labelled solasodine galactoside was observed when UDP-l14]glucose was replaced with UDP-[l4]galactose.
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p.43-53,fig.
Twórcy
autor
- Warsaw University, Al.F.Zwirki i S.Wigury 93, 02-089 Warsaw, Poland
autor
autor
Bibliografia
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- 3. Friedman, M., Henika, P.R. & Mackey, B.E. (1996) Feeding of potato, tomato and eggplant alkaloids affects food consumption and body and liver weights in mice. J. Nutr. 126, -999.
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- 6. Valkonen, J.P.T., Keskitalo, M., Vasara, T. & Pietila, L. (1996) Potato glycoalkaloids: A burden or a blessing? Crit. Rev. Plant Sci.15, 1-20.
- 7. Osbourn, A. (1996) Saponins and plant defence — a soap story. Trends Plant Sci.1,4-9.
- 8. Heftmann, E. (1983) Biogenesis of steroids in Solanaceae. Phytochemistry22, 1843-1860.
- 9. Zimowski, J. (1991) Occurrence of a glucosyltransferase specific for solanidine in potato plants. Phytochemistry30, 1827-1831.
- 10. Stapleton, A., Allen, P.V., Friedman, M. & Belknap, W.R. (1991) Purification and characterization of solanidine glucosyltransferase from the potato (Solanum tuberosum). J. Agric. Food Chem. 39,1187-1193.
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- 12. Zimowski, J. (1992) Specificity and some other properties of cytosolic membranous UDPGlc : 3ß-hyglucosyltransferase from Solanum tuberosumleaves. Phyto- chemistry31, 2977-2981.
- 13. Stapleton, A., Allen, P.V., Tao, H.P., Belknap, W.R. & , M. (1992) amino acid sequence of potato solanidine UDP-glucose glucosyltransferase purified by new an- ion-exchange and size exclusion media. Protein Expression & Purification3, 85-92.
- 14. Zimowski, J. (1994) of UDP- galactose : galactosyltransferase from tomato (Lycopersicon esculentum) . Acta Biochim. Polon.41, 201-204.
- 15. P^czkowski, C. & Wojciechowski, .A (1993) in vitrosynthesis of diosgenin mono- and diglucoside by enzyme preparations from Solanum melongenaleaves. Acta Biochim. Polon.40, 141-143.
- 16. P^czkowski, C. & Wojciechowski, .A. (1994) and galactosylation of diosgenin and solasodine by soluble glucosyltrans- ferase(s) from Solanum melongenaleaves. Phytochemistry35, 1429-1434.
- 17. Kintia, P.K. & , S.A. (1985) gosides N, O and P: Steroidal saponins from seeds of Solanum melongena. Phytochemistry 24, 1567-1569.
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- 21. Pączkowski, . & Wojciechowski, ZA (1988) occurrence of UDPG-dependent glucosyltransferase specific for sarsasapogenin in Asparagus officinalis. Phytochemistry27,2743- 2747.
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- 25. Coves, J., Joyard, J. & , R. (1988) requirement and kinetic studies of solubilized UDP-galactose: galactosyltransferase activity from spinach chloroplast velope membranes. Proc. Natl. Acad. Sci. U.S.A. 85, 4966—4970.
- 26. Heuer, S.fVogt, T., Bóhm, . & , D. (1996) purification and characterization of UDP-glucose :betanidin 5-0- and 6-O-glucosyltransferases from cell suspension cultures of Dorotheanthus bellidiformis. Planta199, 244-250.
- 27. Bechthold, A, Berger, U. & , L. (1991) purification, properties and kinetic studies of UDP-glucose :p-hydroxybenzoate glucosyltransferase from cell cultures of Li- thospermum erythrorhizon. Arch. Biochem. Biophys.288, 38-47.
- 28. Franz, G. (1982) of heterosides (glycosides); in Encyclopedia of Plant Physiology, New senes, . 13A (Loevus, F.A. & , W., eds.) pp. 384-393, Verlag, Berlin.
- 29. Sutter, A & , H. (1975) reversibility of the UDP-glucose: 3-O-gluco- syltransferase reaction. Arch. Biochem. Biophys.167, 444-447.
- 30. Dahlquist, A., Andersson, S. & Wicslander, A. (1992) The enzymatic synthesis of membrane glucolipids in Acholeplasma laidlavii. Bio- chim. Biophys. Acta1105, 131-137.
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- 32. Schmid, G. & Grisebach, H. (1982) Enzymic synthesis of lignin precursors. Purification and properties of UDP-glucose: coniferyl alco- hoi glucosyltransferase from cambial sap of spruce (Picea abiesL.). Eur. J. Biochem.123, 367-370.
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Bibliografia
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