[en] Myrosinase from Brevicoryne brassicae was purified by ammonium sulfate fractionation, dialysis, and chromatography on a DEAE column. The chromatography yielded a single peak and a 115.6-fold purification. Further FPLC gel filtration gave a single peak at 120 kDa. Denaturing SDS/PAGE of the protein revealed a single band at 60 kDa, indicating that the native B. brassicae myrosinase is a dimer. Kinetic parameters towards 8 glucosinolates were calculated. Strong differences of V-max and Km were observed depending on the substrate. Degradation products of each glucosinolate were identified and quantified by GC-MS and GLC-FID, respectively. Using both crude aphid homogenates and purified myrosinase, two unique hydroxyglucosinolates, 3-butenyl- and benzyl-isothiocyanates were identified from progoitrin ((2S)-2-hydroxybut-3-enyl-glucosinolate) and sinalbin (4-hydroxybenzyl-glucosinolate) degradation respectively. Addition of ascorbic acid to the reaction mixtures containing sinalbin and progoitrin caused the production of hydroxyloted degradation products usually associated with plant myrosinase metabolisation. The occurrence of the myrosinase system in B. brassicae is discussed in terms of similar allelochemical adaptation between the herbivore and its host plant. (C) 2002 Wiley-Liss, Inc.
Disciplines :
Anatomy (cytology, histology, embryology...) & physiology Entomology & pest control Biochemistry, biophysics & molecular biology
Bartlet RJ, Mikolajczak KL. 1989. Toxicity of compounds derived from Limnanthes alba seed to fall armyworm (Lepidoptera: Noctuidae). J Econ Entomol 82:1054-1060.
Benn M. 1977. Glucosinolates. Pure Appl Chem 49:197-210.
Birch ANE, Griffiths DW, Mac Farlane Smith WH. 1990. Changes in forage and oilseed rape (B. napus). Root glucosinolates in response to attack by turnip root fly (Delia floralis). J Sci Food Agric 51:309-320.
Björkman R. 1976. Plant myrosinase. In: Vaughan JG, MacLeod AJ, Jones BMG, editors. The biology and chemistry of cruciferae. London: Academic Press, p 191-205.
Björkman R, Janson JC. 1972. Studies on myrosinase. I. Purification and characterization of a myrosinase from a white mustard seed (Sinapis alba L.). Biochim Biophys Acta 276:508-518.
Bones AM, Rossiter JT. 1996. The myrosinase-glucosinolate system, its organisation and biochemistry. Physiol Plant 97:194-208.
Cole R. 1980. Volatile components produced during ontogeny of some cultivated crucifers. J Sci Food Agric 31:549-557.
Durham PL, Poulton JE. 1989. Effect of castanospermine and related polyhydroxyalkaloids on purified myrosinase from Lepidium sativum seedlings. Plant Physiol 90:48-52.
Ehrlick PR, Raven PH. 1964. Butterflies and plants: a study in co-evolution. Evolution 18: 586-608.
Ettlinger MG, Dateo GP, Harrison BW, Mabry TJ, Thompson CP. 1961. Vitamin C as coenzyme : the hydrolysis of mustard oil glucosides. Proc Natl Acad Sci USA 47:1875-1880.
Fenwick GR, Heaney RK, Mullin WJ. 1983. Glucosinolates and their breakdown products in food and food plants. Crit Rev Food Sci Nutr 18:123-201.
Francis F, Haubruge E, Caspar C. 2000. Influence of host plants on the development of the specialist / generalist aphid and on their natural predator Adalia bipunctata. Eur J Entomol 97:481-485.
Francis F, Lognay G, Wathelet JP, Haubruge E. 2001. Effects of allelochemicals from first (Brassicaceae) and second (Myzus persicae & Brevicoryne brassicae) trophic levels on Adalia bipunctata. J Chem Ecol 27:243-256.
Francis F. 1999. Conséquences évolutives des relations entre le puceron et son prédateur en présence de substances allélochimiques chez les Brassicacae. Annales ANPP 2:503-510.
Grob K, Matile P. 1979. Vacuolar location of glucosinolate in horseradish root cells. Plant Sci Lett 14:327-335.
Halkier BA, Du L. 1997. The biosynthesis of glucosinolates. Trends Plant Sci 2:425-431.
Heie OE. 1992. Fauna Entomologica Scandinavica 25, The aphidoidea of Fennoscandia and Denmark. IV. Leiden: Scandinavian Science Press Ltd. 189 p.
Hodek I, Honek A. 1996. Ecology of Coccinellidae. Dordrecht: Kluwer Academic Publisher, 464 p.
Jones AME, Bridges M, Bones AM, Cole R, Rossiter JT. 2001. Purification and characterisation of a non-plant myrosinase from the cabbage aphid Brevicoryne brassicae (L.). Insect Biochem Mol Biol 31:1-5.
Laemmli UK. 1970. Cleavage of structural proteins during assembly of head bacteriophage T4. Nature (Lond) 227: 680-695.
Lamb RJ. 1989. Entomology of oilseed Brassica crops. Annu Rev Entomol 34:211-229.
Lonnerdal B, Janson JC. 1972. Studies on myrosinases. II. Purification and characterization of a myrosinase from a rapeseed (Brassica napus L.). Biochim Biophys Acta 315: 421-429.
Lowry OH, Rosebrough NT, Farr AL, Randhall RT. 1951. Protein measurement with the folin phenol reagent. J Biol Chem 193:265-275.
MacGibbon DB, Allison RM. 1970. A method for the separation and detection of plant glucosinolases (myrosinases). Phytochemistry 9:541-544.
MacGibbon DB, Beuzenberg EJ. 1978. Location of glucosinolate in Brevicoryne brassicae and Lypaphis erysimi (Aphididae). NZ J Sci 21:389-392.
Ohtsuru M, Kawatani H. 1979. Studies of the myrosinase from Wasabia japonica: purification and some properties of wasabi myrosinase. Agric Biol Chem 43:2249-2255.
Othsuru M, Hata T. 1973. Studies on the activation mechanism of the myrosinase by L-ascorbic acid. Agric Biol Chem 37:2543-2548.
Palmieri S, Iori R, Leoni O. 1986. Myrosinase from Sinapis alba L.: A new method of Purification for glucosinolate analyses. J Agric Food Chem 34:138-140.
Pessina A, Thomas R, Palmieri S, Luisi PL. 1990. An improved method for the purification of myrosinase and its physicochemical characterization. Arch Biochem Biophys 280: 383-389.
Porter JRM, Morton AM, Kiddle G, Doughty KJ, Wallsgrove RM. 1991. Variation in the glucosinolates content of oil-seed rape (Brassica napus L.) leaves. Effect of leaf age and position. Ann Appl Biol 118:461-467.
Shikita M, Fahey JW, Golden TR, Holtzclaw WD, 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 41:725-732.
Sones K, Heaney RK, Fenwick GR. 1984. The glucosinolate content of UK vegetables-cabbage (Brassica oleracea), swede (B. napus) and turnip (B. campestris). Food Addit Contam 1:289-296.
Tani N, Othsuru M, Hata T. 1974. Isolation of myrosinase producing microorganism. Agric Biol Chem 38:1617-1622.
Visentin M, Tava A, Iori R, Palmieri S. 1992. Isolation and identification of tran5-4(methylthio)-3-butenyl glucosinolate from radish root (Raphanus sativus). J Agric Food Chem 40:1687-1691.
Wadleigh RW, Yu SJ. 1988. Metabolism of an organothiocyanate allelochemical by glutathione transferase in three lepidopterous insects. J Econ Entomol 81:776-780.
Yen GC, Wei QK. 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.