Development of biocatalysts immobilized on coal ash-derived Ni-zeolite for facilitating 4-chlorophenol degradation.

A new type of biocatalyst was developed to facilitate the biochemical decomposition of 4-chlorophenol (4-CP) in this study. Oxydoreductases that catalyze the initial steps of 4-CP biodegradation were immobilized on a synthetic inorganic enzyme support. Type-X zeolite, a high-surface area support, was synthesized from coal fly ash, on which nickel ions were plated by impregnation (Ni-zeolite), followed by the effective immobilization (77.5% immobilization yield) of recombinant monooxygenase (CphC-I), dioxygenase (CphA-I), and flavin reductase (Fre) isolated from Pseudarthrobacter chlorophenolicus A6 and Escherichia coli K-12, respectively. The retained catalytic activity of the enzymes immobilized on Ni-zeolite was as high as 64% of the value for the corresponding free enzymes. The Michaelis-Menten kinetic parameters v and K of the immobilized enzymes were determined to be 0.20 mM·min and 0.44 mM, respectively. These results are expected to provide useful information with respect to the development of novel enzymatic treatments for phenolic hydrocarbon contaminants.