Add time:09/02/2019         Source:sciencedirect.com

2,4-Dichlorophenoxyacetic acid (2,4-D)/α-ketoglutarate (α-KG) dioxygenase, TfdA, from Ralstonia eutropha JMP134, was purified from recombinant cells and shown by gas chromatographic and colorimetric methods to degrade only the S enantiomer of dichlorprop, a phenoxypropionate herbicide. Similarly, cell extracts of Burkholderia cepacia RASC, containing a biochemically and genetically related α-KG-dependent dioxygenase, also were shown to oxidize (S)-dichlorprop using chiral HPLC and colorimetric methods. In contrast, cell extracts of a mecoprop-degrading strain of Alcaligenes denitrificans were shown to catabolize (R)-dichlorprop. Although the A. denitrificans activity exhibited stereospecificity opposite to that of the JMP134 and RASC strains, its cofactor requirements were found to be characteristic of an α-KG-dependent dioxygenase. A PCR amplification product from the DNA of this strain was shown to encode an amino acid sequence that was 95% and 86% identical to the corresponding region of TfdA in RASC and JMP134, respectively. Thus, closely related herbicide-degrading gene products appear to be capable of exhibiting opposite stereochemical degradative capabilities.

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