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Method to Determine Guaiacol Oxidase Activity in Soil

Dipartimento di Scienza del Suolo e Nutrizione della Pianta, P.le delle Cascine 15, 50144 Firenze, Italy

Antonio Gelsomino

Dipartimento di Agrobiologia ed Agrochimica, Universita' della Tuscia, Via S.C. De Lellis, 01100 Viterbo, Italy

Marcello Felici

Instituto di Chimica Agraria e Forestale, Piazza S. Francesco, 89061 Gallina di Reggio Calabria, Italy

*Corresponding author.

ABSTRACT

A precise and sensitive method to assay guaiacol oxidase activity in soil is needed. The objective of this study was to set up this method by determining residual guaiacol spectrophotometrically after its extraction from soil with 0.5 M NaOH and 1 M CaCl₂ and its separation from reaction products and humic molecules of the soil extract by high-performance liquid chromatography (HPLC). A soil sample sterilized by autoclaving was used as a control. The rate of guaiacol oxidation reached the highest value at pH 4.5 to 6.0. The reaction rate of the enzyme approached zero-order kinetics when the substrate concentration was 50 mM. Disappearance of guaiacol was linear with time for at least 1 h. The optimal temperature for guaiacol oxidation activity occurred at 60°C. Guaiacol oxidase activity was significantly correlated with soil total N.

Received for publication May 14, 1990.