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TECHNICAL REPORTS
Bioremediation and Biodegradation

Treatment of 2,4-Dichlorophenol Polluted Soil with Free and Immobilized Laccase

^a Laboratory of Soil Biochemistry, The Pennsylvania State University, 129 Land and Water, University Park, PA 16802
^b Dep. of Agricultural Chemistry, Kyungpook National University, Taegu, 702-701, Korea

* Corresponding author (jmbollag{at}psu.edu)

Received for publication October 8, 2001. Enzyme treatment is currently considered for remediation of terrestrial systems polluted with organic compounds. In this study, two soils from Pennsylvania with 2.8 or 7.4% organic matter contents (Soils 1 and 2, respectively) were amended with ¹⁴C-labeled 2,4-dichlorophenol (2,4-DCP) and incubated with a laccase from Trametes villosa (free or immobilized on montmorillonite). 2,4-DCP was either transformed to methanol-soluble polymeric products (11–32%) or covalently bound to soil organic matter (53–85%); unaltered 2,4-DCP could be recovered from soil by methanol extraction (0–38%) at the completion of a 14-d incubation period. In Soil 1, both free and immobilized laccase removed 100% of 2,4-DCP without regard for moisture conditions. In Soil 2, immobilized laccase removed more 2,4-DCP (about 95%, regardless of moisture conditions) than free enzyme (55, 75, and 90% at 30, 55, and 100% of maximum water-holding capacity, respectively). Binding of 2,4-DCP in the humin fraction was nearly the same for free and immobilized laccase. More 2,4-DCP, however, was bound to humic and fulvic acids in the presence of immobilized laccase than in the presence of free laccase. In general, immobilized laccase performed better than free laccase. However, for practical applications, the higher activity of immobilized laccase is offset by a 23% loss in enzyme activity during immobilization, which approximates the 30% increase in free laccase needed to achieve the same level of remediation. Furthermore, immobilized laccase is more costly than free T. villosa laccase.

Abbreviations: 2,4-DCP, 2,4-dichlorophenol • HPLC, high-performance liquid chromatography • WHC, water-holding capacity