The effects of 12 common metal ionic compounds on the laccase catalytic activity in reactions using guaiacol as the substrate was determined using spectrophotometry. Furthermore, the influence of several metal ionic compounds on the generation of reactive oxygen species (ROS) by oxidation of lignin in jute fiber under laccase catalysis was studied by electron paramagnetic resonance (EPR) spectroscopy using N-tert-butyl-alpha-phenylnitrone (PBN) as the spin-trapping agent. Common metal cations, such as K + , Na + , Mg 2+ , Ca 2+ , and Cu 2+ and the anion SO4 2-had almost no effect on laccase activity during the initial stage of the catalytic reactions. High concentrations of the Mn 2+ ion exhibited weak inhibition of laccase; Ag + and NO3-showed a moderate inhibitory effect on laccase activity during the initial stage of the catalytic reactions. Fe 2+ had no direct effect on the binding of laccase to its substrate, but strongly retarded the progress of the catalytic reaction by reducing the intermediate free radicals. The ions Cl-, Fe 3+ , and Ag + exhibited either strong inhibitory effects on the catalysis of the substrate or a destructive effect on the structure of laccase itself. Furthermore, the results showed that an appropriate concentration of Cu 2+ helped to promote the thermal stability of laccase during the enzymatic reaction. This study could help researchers to avoid the use of inhibitory exogenous metal ions and anions in the application of laccase and to maximize the value of laccase.

Arnebia euchroma callus, obtained from the root cell culture of an Iranian native specimen, has gained a doubling time of 63 H after regular subculturing on Linsmaier-Skoog (LS) medium containing sugar (50 g/L), 2,4-dichlorophenoxyacetic acid (10(-6) M), and kinetin (10(-5) M) under darkness at 25°C. Despite the observed somaclonal variations, peroxidase production by the A. euchroma calli has been stable over 4 years under the aforementioned conditions. Isoelectric focusing experiments revealed that the partially purified A. euchroma peroxidases (AePoxs) are mainly anionic with pI values of about 5.5 and 6.6. AePox reaches its optimal activity at 55°C and pH 7.5. Results of the various kinetic studies suggest that AePox belongs to the type III plant peroxidases with no activity for the oxidation of 3-indoleacetic acid, but seems to play a role in the lignin biosynthesis and H(2) O(2) regulation during the proliferation of the A. euchroma cells on LS medium. Comparing the biochemical properties of AePox with horseradish peroxidase and in view of the ease of solid cell culture, the A. euchroma callus could be considered as a source of plant peroxidase for some biotechnological applications.