For measurement of Guaiacol Peroxidase (GPX), 10 lL of plant extract were added to the reaction mixture (giving a final volume of 3 mL) containing 50 mM phosphate buffer, 20 mM
guiacol (Sigma-Aldrich) and 40 mM H2O2 (pH 7.0; Macron).
Santimone, "Titration study of
guiacol oxidation by horseradish peroxidase," Canadian Journal of Biochemistry, vol.
The running buffer was of Tris Glycene native page of pH 8.9 (Goldenberg 1989).Peroxidase isozymes were localized by incubating gel in
Guiacol reagent (0.25%) for 30 min followed by incubation in 0.3% hydrogen peroxide for another 15 min.
[23] reported an optimum temperature of 30[degrees]C for peroxidase from Turkish black radish using
guiacol as substrate.
A 1 ml reaction mixture consisted of 28 mM
guiacol buffer, 5 mM [H.sub.2][O.sub.2] buffer, 20 mM Tris-maleate buffer and the enzyme extract.
He had been working with guiacol, a compound that was found in creosote, the black oily guck that builds up inside chimney flues as a result of incomplete burning of wood or coal.
As part of his research, Fox synthesized a number of guiacol derivatives, one of which was guiacol carbonate.
Guiacol 99%, l-naphthol 98%, o-cresol 98%, and dichloromethane 99.5% were purchased from SRL (India).
The reaction of substituted phenols (o-cresol, guiacol, phenyl phenol, l-naphthol, and 2-allyl phenol) with thionyl chloride in the presence of anhydrous aluminum chloride in dichloromethane yields sulfoxide-bridged bisphenols (13), (14).
The reaction mixture was prepared with 0.05m1 of 20mM
guiacol, 3m1 of phosphate buffer, O.lml of enzyme extract and 0.03m1 of [H.sub.2][O.sub.2].
We will demonstrate and describe the procedures and teaching techniques developed at an NSF-funded summer workshop for a general university biology laboratory to determine the optimum pH for a peroxidase and
guiacol reaction.