In this study an oxidative enzyme was immobilized onto inorganic backbone materials to stimulate the detoxification of toxic aromatic hydrocarbon compounds. Smectite clay minerals and soil organic matter were screened as an enzyme support and a binding agent, respectively. Montmorillonite of which inner pores are layered with nano-scale spacing planar was activated by humic acid. A dioxygenase obtained by cloning of its corresponding gene from Arthrobacter chlorophenolicus A6 was immobilized onto the humic acid-activated montmorillonite. Oxygenated metabolites such as catechol and 4-chlorocatechol were selected as target aromatic contaminants (primary substrates of enzyme). The enzyme immobilization yield was as high as 63% and the reductions in enzyme activity for the decomposition of substrate compounds during enzyme immobilization were minimal: 15% for catechol and 24% for 4-chlorocatechol, respectively. The kinetic analysis of the free and immobilized enzymes demonstrated a slight decrease of vmax and a marginal increase of KM as compared with those for the free enzyme, indicating the changes in enzyme activity perhaps due to the changes in enzyme conformation associated with its immobilization were minimal. The results for the effects of environmental factors including pH, temperature, and ionic strength on the activity of free and immobilized enzymes showed that the activity of free enzyme changed significantly in response to the changes of the environmental factors whereas that of immobilized enzyme was pretty much consistent. This indicated that the stability of enzyme against the abrupt changes in environmental factors can be greatly improved by enzyme immobilization. The results of this study support the feasibility of a new environmental fusion technology based on bio-technology and nano-technology for the development of biochemical treatment processes.