metabolizes testosterone as the sole carbon source via a cleavage reaction. A -cleavage enzyme gene, , was cloned from TA441. The deduced N-terminal amino acid sequence of matched that of the purified -cleavage enzyme which is induced in TA441 during growth on testosterone as the sole carbon source. The disrupted mutant did not show growth on testosterone, suggesting that is necessary for TA441 to grow on testosterone. Downstream from , three putative ORFs which encode products also necessary for growth of TA441 on testosterone were identified. The usual substrate of TesB is probably 3,4-dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione. Although this compound was not identified in the gene disrupted mutants, accumulation of upstream metabolites of testosterone degradation, 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione, was shown by TLC analysis.


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