Activity of myrosinase from Sinapis alba seeds immobilized into Ca-polygalacturonate as a simplified model of soil-root interface mucigel

作者：Braschi Ilaria^*; Leoni Onofrio; Cinti Susanna; Palmieri Sandro; Gessa Carlo Emanuele

来源：Plant and Soil, 2011, 339(1-2): 209-218.

DOI：10.1007/s11104-010-0569-x

• Myrosinase
• Immobilization
• Glucosinolates
• Pectins
• Root mucigel
• Calcium polygalacturonate

Ca-polygalacturonate is a demethoxylated component of pectins which are constitutive of plant root mucigel. In order to define the role of root mucigel in myrosinase immobilization and activity at root level, a myrosinase enzyme which had been isolated from Sinapis alba seeds was immobilized into Ca-polygalacturonate. The activity profile for the immobilized and free enzyme was evaluated using the pH-Stat method as a function of time, temperature, and pH. The Michaelis-Menten kinetic parameters change between the immobilized (V (max) = 127 +/- 13 U mg(-1) protein; K (M) = 6.28 +/- 0.09 mM) and free (V (max) = 17 +/- 1 U mg(-1) protein; K (M) = 0.96 +/- 0.01 mM) forms of myrosinase, probably due to conformational changes involving the active site as a consequence of enzyme immobilization. Immobilized enzyme activity evaluated as a function of different substrates gave the highest value with nasturtin, the glucosinolate that is typical of several brassicaceae plant roots containing the glucosinolate-myrosinase defensive system. No feedback regulation mechanism was found in the presence of an excess of enzymatic reaction products (i.e. allyl isothiocyanate or sulphate). The high enzyme immobilization yield into Ca-polygalacturonate and its activity preservation under different conditions suggest that the enzyme released by plants at root level could be entrapped in root mucigel in order to preserve its activity.

• 出版日期2011-2