Cultures of the pyrene degrading Mycobacterium sp. KMS were incubated with [4-13C]pyrene or [4,5,9,10-14C]pyrene with and without a soil humic acid standard to characterize the chemical nature of the produced residues and evaluate the potential for bonding reactions with humic acid. Cultures were subjected to a “humic acid/humin” separation at acidic pH, a duplicate separation followed by solvent extraction of the humic acid/humin fraction, and a high pH separation. 13C NMR analysis was conducted on the resulting solid extracts. Results indicated that the activity associated with solid extracts did not depend on pH and that approximately 10% of the added activity was not removed from the solid humic acid/humin fraction by solvent extraction. 13C NMR analysis supported the conclusion that the majority of pyrene metabolites were incorporated into cellular material. Some evidence was found for metabolite reaction with the added humic material, but this did not appear to be a primary fate mechanism.