Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods - Publication - Bridge of Knowledge

Search

Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods

Abstract

Myrosinase found in Brassicaceae plants, is the enzyme responsible for hydrolysis of glucosinolates. As a result a variety of biologically active metabolites are liberated, whose importance in crop protection and especially in cancer chemoprevention is rapidly gaining recognition. The growing practical application of glucosinolate degradation products requires that sensitive and reliable methods of myrosinase activity determination in different types of plant samples are established. With the use of commercial myrosinase prep, we systematically optimised conditions of measurement of this enzyme activity by spectrophotometric and pH-stat methods. The parameters evaluated included: sample preparation, choice of substrate, its concentration, reaction temperature and detection wavelength. Two substrates with different spectral properties were chosen: sinigrin (SIN) and glucotropaeolin (GTL). For both substrates, the best reliability was achieved at reaction temperature of 37*C and substrate concentration of 0,2mM and 5mM for spectrophotometric and pH-stat methods, respectively. GTL exhibiting higher absortpion at the recommended detection wavelenght of 230 nm ensured greater sensitivity of spectrophotometric determination of myrosinase activity in the case of transparent plan samples. GTL seemed to increase also the sensitivity of pH-stat method, however, in this casehomogenisation of plant samples turned out to be most important. The optimised conditions were then verified for a range of plant samples. Based on these results, the optimised protocols of myrosinase activity determination for both methods are proposed.

Citations

  • 2 4

    CrossRef

  • 0

    Web of Science

  • 2 6

    Scopus

Cite as

Full text

download paper
downloaded 979 times
Publication version
Accepted or Published Version
License
Creative Commons: CC-BY-NC-ND open in new tab

Keywords

Details

Category:
Articles
Type:
artykuł w czasopiśmie wyróżnionym w JCR
Published in:
INDUSTRIAL CROPS AND PRODUCTS no. 50, pages 58 - 67,
ISSN: 0926-6690
Language:
English
Publication year:
2013
Bibliographic description:
Piekarska A., Kusznierewicz B., Meller M., Dziedziul K., Namieśnik J., Bartoszek-Pączkowska A.: Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods// INDUSTRIAL CROPS AND PRODUCTS. -Vol. 50, (2013), s.58-67
DOI:
Digital Object Identifier (open in new tab) 10.1016/j.indcrop.2013.06.048
Bibliography: test
  1. Anderton, M., Jukes, R., Lamb, J., Manson, M., Gescher, A., Steward, W., Williams, M., 2003. Liquid chromatographic assay for simultaneous determination of indole-3-carbinol and its condensation products in plasma. J. Chromatogr. B 787, 281-291. open in new tab
  2. Andreasson, E., Jorgensen, L.B., Hoeglund, A.S., Rask, L., Meijer, J., 2001. Different myrosinase and idioblast distribution in Arabidopsis and Brassica napus. Plant Physiol. 127, 1750-1763. open in new tab
  3. Bellostas, N., Petersen, I.L., Sorensen, J.C., Sorensen, H., 2008a. A fast and gentle method for the isolation of myrosinase complexes from Brassicaceous seeds. J. Biochem. Biophys. Methods 70, 918-925. open in new tab
  4. Bellostas, N., Sørensen, A.D., Sørensen, J.C., Sørensen, H., 2008b. Fe 2+ -catalyzed for- mation of nitriles and thionamides from intact glucosinolates. J. Nat. Prod. 71, 76-80. open in new tab
  5. Bernardi, R., Finiguerra, M.G., Rossi, A.A., Palmieri, S., 2003. Isolation and biochemical characterization of a basic myrosinase from ripe Crambe abyssinica seeds, highly specific for epi -progoitrin. J. Agric. Food Chem. 51, 2737-2744. open in new tab
  6. Björkman, R., Janson, J., 1972. Studies on myrosinase 1. Purification and character- ization of a myrosinase from white mustard seeds (Sinapsis alba L). Biochim. Biophys. Acta 276, 508-518. open in new tab
  7. Björkman, R., Lönnerdal, B., 1973. Studies on myrosinase 3: enzymatic properties of myrosinases from Sinapsis alba and Brassica napus seeds. Biochim. Biophys. Acta 327, 121-131. open in new tab
  8. Bodnaryk, R.P., 1991. Developmental profile of sinalbin (p-hydroxybenzyl glucosino- late) in mustard seedlings, Sinapis alba L, and its relationship to insect resistance. J. Chem. Ecol. 17, 1543-1556. open in new tab
  9. Bones, A., Iversen, T., 1985. Myrosin cells and myrosinase. Isr. J. Bot. 34, 351-376. open in new tab
  10. Burow, M., Bergner, A., Gershenzon, J., Wittstock, U., 2007. Glucosinolate hydrolysis in Lepidium sativum--identification of the thiocyanate-forming protein. Plant Mol. Biol. 63, 49-61. open in new tab
  11. Charron, C.S., Saxton, A.M., Sams, C.E., 2005. Relationship of climate and genotype to seasonal variation in the glucosinolate -myrosinase system II. Myrosinase activity in ten cultivars of Brassica oleracea grown in fall and spring seasons. J. Sci. Food Agric. 85, 682-690. open in new tab
  12. Davey, M.W., Montagu, M., Inze, D., Sanmartin, M., Kanellis, A., Smirnoff, N., Benzie, I.J.J., Strain, J.J., Favell, D., Fletcher, F., 2000. Plant l-ascorbic acid: chemistry, function, metabolism, bioavailability and effects of processing. J. Sci. Food Agric. 80, 825-860. open in new tab
  13. Durham, P., Poulton, J., 1990. Enzymatic properties of purified myrosinase from Lepidium sativum seedlings. Z. Naturforch. C Biosci. 45, 173-178. open in new tab
  14. Eriksson, S., Andreasson, E., Ekbom, B., Granér, G., Pontoppidan, B., Taipalensuu, J., Zhang, J., Rask, L., Meijer, J., 2002. Complex formation of myrosinase isoenzymes in oilseed rape seeds are dependent on the presence of myrosinase-binding protein. Plant Physiol. 129, 1592-1599. open in new tab
  15. Fahey, J., Zalcmann, A., Talalay, P., 1997. Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. Proc. Natl. Acad. Sci. 94, 10367-10372. open in new tab
  16. Finiguerra, M.G., Iori, R., Palmieri, S., 2001. Soluble and total myrosinase activity in defatted Crambe abyssinica meal. J. Agric. Food Chem. 49, 840-845. open in new tab
  17. Gimsing, A., Kirkegaard, J.A., 2009. Glucosinolates and biofumigation: fate of glu- cosinolates and their hydrolysis products in soil. Phytochem. Rev. 8, 299-310. open in new tab
  18. Hoglund, A.S., Lenman, M., Rask, L., Rask, A., 1991. Distribution of myrosinase in rapeseed tissues. Plant Physiol. 95, 213-221. open in new tab
  19. Johnson, I., 2002. Glucosinolates bioavailabity and importance to health. Int. J. Vitam. Nutr. Res. 72, 26-31. open in new tab
  20. Kirkegaard, J.A., Sarwar, M., 1998. Biofumigation potential of brassicas. Plant Soil 201, 71-89. open in new tab
  21. Kleinwächter, M., Selmar, D., 2004. A novel approach for reliable activity determi- nation of ascorbic acid depending myrosinases. J. Biochem. Biophys. Methods 59, 253-265. open in new tab
  22. Kusznierewicz, B., Iori, R., Piekarska, A., Namieśnik, J., Bartoszek, A., 2013. Con- venient identification of desulfoglucosinolates on the basis of mass spectra obtained during liquid chromatography-diode array-electrospray ionization mass spectrometry analysis: method verifications for sprouts of different Brassicaceae species extracts. J. Chromatogr. A, http://dx.doi.org/10.1016/j. chroma.2012.12.075. open in new tab
  23. Larkin, R., Griffin, T., 2007. Control of soilborne potato diseases using Brassica green manures. Crop Prot. 26, 1067-1077. open in new tab
  24. Leroux, M., Foster-Powell, K., Holt, S., Brand-Miller, J., 2002. International table of glycemic index and glycemic load values. Am. J. Clin. Nutr. 76, 5-56.
  25. Li, X., Kushad, M.M., 2005. Purification and characterization of myrosinase from horseradish (Armoracia rusticana) roots. Plant Physiol. Biochem. 43, 503-511. open in new tab
  26. Manici, L., Lazzeri, L., Palmieri, S., 1997. In vitro fungitoxic activity of some glucosi- nolates and their enzyme-derived products toward plant pathogenic fungi. J. Agric. Food Chem. 45, 2768-2773. open in new tab
  27. Mithen, R., 2001. Glucosinolates -biochemistry genetics and biological activity. Plant Growth Regul. 34, 91-103. open in new tab
  28. Palmieri, S., Iori, R., Leoni, O., 1987. Comparison of methods for determining myrosi- nase activity. J. Agric. Food Chem. 35, 617-621. open in new tab
  29. Pocock, K., Heaney, R.K., Wilkinson, A.P., Beaumont, J.E., Vaughan, J.G., Fenwick, G.R., 1987. Changes in myrosinase activity and isoenzyme pattern, glucosinolate con- tent and the cytology of myrosin cells in the leaves of heads of three cultivars of English white cabbage. J. Sci. Food Agric. 41, 245-257. open in new tab
  30. Shapiro, T.A., Fahey, J.W., Wade, K.L., Stephenson, K.K., Talalay, P., 2001. Chemopro- tective glucosinolates and isothiocyanates of broccoli sprouts: metabolism and excretion in humans. Cancer Epidemiol. Biomarkers Prev. 10, 501-508. open in new tab
  31. Shikita, M., Fahey, J.W., Golden, T., Holtzclaw, W., Talalay, P., 1999. An unusual case of 'uncompetitive activation' by ascorbic acid: purification and kinetic properties of a myrosinase from Raphanus sativus seedlings. Biochem. J. 341, 725-732. open in new tab
  32. Singh, J., Rai, M., Upadhyay, A.K., Prasad, K., 2007. Sinigrin (2-propenyl glucosino- late) content and myrosinase activity in Brassica vegetables. Int. J. Veget. Sci. 13, 21-31. open in new tab
  33. Smith, T.K., Mithen, R., Johnson, I.T., 2003. Effects of brassica vegetable juice on the induction of apoptosis and aberrant crypt foci in rat colonic mucosal crypts in vivo. Carcinogenesis 24, 491-495. open in new tab
  34. Thangstad, O., Iversen, T., Slupphaug, G., Bones, A., 1990. Immunocytochemical local- ization of myrosinase in Brassica napus L. Planta 180, 245-248. open in new tab
  35. Travers-Martin, N., Kuhlmann, F., Muller, C., 2008. Revised determination of free and complexed myrosinase activities in plant extracts. Plant Physiol. Biochem. 46, 506-516. open in new tab
  36. Vallejo, F., Thomas-Barberan, F., Garcia-Viguera, C., 2003. Health-promoting com- pounds in broccoli as influenced by refrigerated transport and retail sale period. J. Agric. Food Chem. 51, 3029-3034. open in new tab
  37. Wilkinson, A.P., Rhodes, M.J.C., Fenwick, R.G., 1984. Myrosinase activity of crucifer- ous vegetables. J. Sci. Food Agric. 35, 543-552. open in new tab
  38. Wu, L., Ashraf, M.H., Facci, M., Wang, R., Paterson, P.G., Ferrie, A., 2004. Dietary approach to attenuate oxidative stress, hypertension, and inflammation in the cardiovascular system. Proc. Natl. Acad. Sci. U.S.A. 101, 7094-7099. open in new tab
  39. Yen, G-C., Wei, Q-K., 1993. Myrosinase activity and total glucosinolate content of cruciferous vegetables, and some properties of cabbage myrosinase in Taiwan. J. Sci. Food Agric. 61, 471-475. open in new tab
  40. Zhang, Z., Ober, J., Kliebenstein, D., 2006. The gene controlling the quantitative trait locus epithiospecifier modifier 1 alters glucosinolate hydrolysis and insect resis- tance in Arabidopsis. Plant Cell 18, 1524-1536. open in new tab
Verified by:
Gdańsk University of Technology

seen 202 times

Recommended for you

Meta Tags