Transformation of tetracycline by TetX and its subsequent degradation in a heterologous host

TetX is a flavin-dependent monooxygenase. It has been reported to inactivate all tested tetracyclines. In this study, Escherichia coli overexpressing TetX was added to soil bacterial enrichment cultures along with varying levels of tetracycline and was found to affect community-wide tetracycline resistance levels. Soil microbial communities developed lower levels of tetracycline resistance upon exposure to 25μg/mL of tetracycline when an E. coli expressing TetX was present (6% of cultivable bacteria were resistant to 40μg/mL tetracycline). In contrast, in the absence of TetX activity, a similar tetracycline exposure selected for greater levels of resistant bacteria in the soil microbial community (90% of cultivable bacteria were resistant to 40μg/mL tetracycline). We also describe new metabolites formed after tetracycline transformation by TetX and report the transient generation of redox-active metabolite(s). The results presented here are particularly pertinent in the light of the recent emergence of tet(X) in different bacterial species, including clinical isolates.