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

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Abstract

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.

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Abbreviations

MYR:

myrosinase

GLs:

glucosinolates

PGA:

polygalacturonic acid

Ca-PG:

Calcium polygalacturonate

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Acknowledgement

The authors wish to acknowledge Dr. Antonio E. Faleo for his laboratory assistance.

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Authors and Affiliations

  1. Dipartimento di Scienze e Tecnologie Agroambientali, Università di Bologna, Viale Fanin, 44-40127, Bologna, Italy

    Ilaria Braschi & Carlo Emanuele Gessa

  2. Centro di Ricerca per le Colture Industriali CRA-CIN, Via di Corticella, 133-40128, Bologna, Italy

    Onofrio Leoni, Susanna Cinti & Sandro Palmieri

Authors
  1. Ilaria Braschi
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  2. Onofrio Leoni
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  3. Susanna Cinti
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  4. Sandro Palmieri
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  5. Carlo Emanuele Gessa
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Correspondence to Ilaria Braschi.

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Responsible Editor: Harsh P. Bais.

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Braschi, I., Leoni, O., Cinti, S. et al. Activity of myrosinase from Sinapis alba seeds immobilized into Ca-polygalacturonate as a simplified model of soil-root interface mucigel. Plant Soil 339, 209–218 (2011). https://doi.org/10.1007/s11104-010-0569-x

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