Multicomponent kinetic analysis and theoretical studies on the phenolic intermediates in the oxidation of eugenol and isoeugenol catalyzed by laccase.

Laccase catalyzes the oxidation of natural phenols and thereby is believed to initialize reactions in lignification and delignification. Numerous phenolic mediators have also been applied in laccase-mediator systems. However, reaction details after the primary O-H rupture of phenols remain obscure. In this work two types of isomeric phenols, EUG (eugenol) and ISO (trans-/cis-isoeugenol), were used as chemical probes to explore the enzymatic reaction pathways, with the combined methods of time-resolved UV-Vis absorption spectra, MCR-ALS, HPLC-MS, and quantum mechanical (QM) calculations. It has been found that the EUG-consuming rate is linear to its concentration, while the ISO not. Besides, an o-methoxy quinone methide intermediate, (E/Z)-4-allylidene-2-methoxycyclohexa-2,5-dienone, was evidenced in the case of EUG with the UV-Vis measurement, mass spectra and TD-DFT calculations; in contrast, an ISO-generating phenoxyl radical, a (E/Z)-2-methoxy-4-(prop-1-en-1-yl) phenoxyl radical, was identified in the case of ISO. Furthermore, QM calculations indicated that the EUG-generating phenoxyl radical (an O-centered radical) can easily transform into an allylic radical (a C-centered radical) by hydrogen atom transfer (HAT) with a calculated activation enthalpy of 5.3 kcal mol(-1) and then be fast oxidized to the observed eugenol quinone methide, rather than an O-radical alkene addition with barriers above 12.8 kcal mol(-1). In contrast, the ISO-generating phenoxyl radical directly undergoes a radical coupling (RC) process, with a barrier of 4.8 kcal mol(-1), while the HAT isomerization between O- and C-centered radicals has a higher reaction barrier of 8.0 kcal mol(-1). The electronic conjugation of the benzyl-type radical and the aromatic allylic radical leads to differentiation of the two pathways. These results imply that competitive reaction pathways exist for the nascent reactive intermediates generated in the laccase-catalyzed oxidation of natural phenols, which is important for understanding the lignin polymerization and may shed some light on the development of efficient laccase-mediator systems.

[Accumulation Characteristics and Evaluation of Heavy Metals in Soil-Crop System Affected by Wastewater Irrigation Around a Chemical Factory in Shenmu County].

Soil heavy metals Cu, Pb, Zn, and Cd, are regarded as "chemical time bombs" because of their propensity for accumulation in the soil and uptake by crops. This ultimately causes human toxicity in both the short and long-term, making farmland ecosystems dangerous to health. In this paper, accumulation and spatial variability of Cu, Zn, Pb and Cd in soil-crop system affected by wastewater irrigation around a chemical factor in northern Shaanxi province were analyzed. Results showed that wastewater irrigation around the chemical factory induced significant accumulation in soils compared with control areas. The average concentrations of available Cu and total Cu were 4.32 mg x kg(-1) and 38.4 mg x kg(-1), which were twice and 1.35 times higher than those of the control area, respectively. Soil Zn and Pb were slightly accumulated. Whereas soil Cd was significantly accumulated and was higher than the critical level of soil environmental quality (II), the available and total Cd concentrations were 0.248 mg x kg(-1) and 1.21 mg x kg(-1), which were 10 and 6.1 times higher than those of the control areas. No significant correlations were found between available and total heavy metals except between available Cd and total Cd. All the heavy metals were mainly accumulated in the top layer (0-10 cm). Spatially, soils and plants high in heavy metal concentration were distributed within the radius of about 100 m from the waste water outlet for Cu, Zn and Cd and about 200 m for Pb, and decreased exponentially with the distance from the factory. Affected by wastewater irrigation, contents of Cu, Pb and Cd in maize were 4.74, 0.129 and 0.036 mg x kg(-1) which were slightly higher than those in the control area. The content of Zn was similar to that in the control area. Affected by the vehicle exhaust, the over standard rate of Pb was 5.7% in maize. All the heavy metals did not show significant correlation between soil and crop, except Cd. The square correlation coefficients were 0.83 and 0.75 between soil available and total Cd with maize. Therefore, the chemical factory contributed to the accumulation of heavy metals in the soil around it, but the contribution to the crop was limited, and thus temporarily caused no threat to human health. The reason for the lower accumulation was the high soil pH and low soil organic matter content. But more attentions should be paid to the higher accumulation of Pb in the maize caused by the vehicle and dust.