A fermentative enrichment culture (designated DHM-1) was developed that is capable of cometabolically biotransforming high concentrations of chloroform (CF) to nontoxic end products. Two Pantoea spp. were isolated from DHM-1 that also possess this dechlorination capability. Following acclimation to increasing levels of CF, corn syrup-grown DHM-1 was able to transform over 500 mg/liter CF in the presence of vitamin B 12 (approximately 3% of CF on a molar basis) at a rate as high as 22 mg/liter/day in a mineral salts medium. CO, CO 2 , and organic acids were the predominant biodegradation products, suggesting that hydrolytic reactions predominate during CF transformation. DHM-1 was capable of growing on corn syrup in the presence of high concentrations of CF (as may be present near contaminant source zones in groundwater), which makes it a promising culture for bioaugmentation. Strains DHM-1B and DHM-1T transform CF at rates similar to that of the DHM-1 enrichment culture. The ability of these strains to grow in the presence of high concentrations of CF appears to be related to alteration of membrane fluidity or homeoviscous and homeophasic adaptation.Chloroform (CF) is a toxic organic compound that is frequently detected in groundwater. In the 2007 "CERCLA Priority List of Hazardous Substances," CF ranks eleventh overall and is the third highest among chlorinated organics after vinyl chloride and polychlorinated biphenyls (4). When present at hazardous waste sites, CF is often a focal point for evaluating the feasibility of bioremediation, since it is toxic to many obligate anaerobic prokaryotes (44). For example, 1 mg/liter of CF completely inhibited dechlorination of tetrachloroethene (PCE) by a chlororespiring anaerobic isolate (32). Inhibition of reductive dechlorination of chloroethenes by CF is a general problem for sites cocontaminated with CF, which can only be overcome by first removing the CF (6).In spite of major recent advances in bioremediation of chlorinated organic compounds, treatment of CF, especially at high concentrations (e.g., Ͼ100 mg/liter), remains challenging. Although aerobic biotransformation of CF is possible (e.g., cometabolism by a butane-grown strain) (14), CF is more difficult to cometabolize than trichloroethene (42). Biotransformation of CF by mixed or pure cultures under methanogenic (5, 21) and sulfate-reducing (20) conditions has been reported, however, only at low-mg/liter CF concentrations.Corrinoids such as vitamin B 12 (i.e., cyanocobalamin) are effective catalysts for increasing the rate of halomethane biotransformation under anaerobic conditions. Addition of vitamin B 12 also shifts the pathway away from reductive dechlorination and toward hydrolytic and substitutive reactions, forming CO, CO 2 , and organic acids as the major products (8,23,24). With low levels of B 12 added (3 to 5% molar ratios of CF), an enrichment culture grown on dichloromethane (DCM) as the sole substrate (8) and a lactate-grown sulfate-reducing enrichment culture (18) were able to biotransform up ...
