E. Dupille et L. Zacarias, EXTRACTION AND BIOCHEMICAL-CHARACTERIZATION OF WOUND-INDUCED ACC OXIDASE FROM CITRUS PEEL, PLANT SCI, 114(1), 1996, pp. 53-60
1-Aminocyclopropane-1-carboxylate (ACC) oxidase from flavedo tissue of
mandarin fruit was extracted and its biochemical properties investiga
ted. Enzyme activity was induced by wounding and exposure to light. In
vitro ACC oxidase activity required the presence of Fe2+ and ascorbat
e and was enhanced by addition of NaHCO3 to the reaction mixture. The
apparent K-m of ACC was estimated to be 175 mu M in 30 mM NaHCO3. Reco
very of the activity was only possible after passing the crude extract
through Sephadex. The fraction retained in the column was found to st
rongly reduce the activity of the enzyme. Hence, the presence of low m
olecular weight inhibitors in the crude extract of mandarin flavedo is
implied. ACC oxidase activity from Citrus has two temperature maxima,
at 35 degrees C and 45 degrees C, and is inhibited by sulfhydryl reag
ents, metal chelators, free radical scavengers and divalent cations. T
hese results indicate that ACC oxidase activity recovered from wounded
flavedo tissue resembles authentic in vivo activity and that the bioc
hemical properties of the enzyme in a non-climacteric fruit are very s
imilar to those observed in other climacteric-like tissues.