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GenParts™ DNA Fragments> | … The terB and terC genes were obtained from synthetic nucleic acids (GenScript). Both genes were digested using NdeI and HindIII restriction enzymes (New England Biolabs). The corresponding fragments were cloned into the E. coli expression vector pET28a (Novagen) and … | Get A Quote |
We report the identification of the gene cluster responsible for the formation of the -terphenyl derivatives terfestatins B and C and echoside B from the Appalachian strain RM-5-8. We characterize the function of TerB/C, catalysts that work together as a dual enzyme system in the biosynthesis of natural terphenyls. TerB acts as a reductase and TerC as a dehydratase to enable the conversion of polyporic acid to a terphenyl triol intermediate. X-ray crystallography of the apo and substrate-bound forms for both enzymes provides additional mechanistic insights. Validation of the TerC structural model via mutagenesis highlights a critical role of arginine 143 and aspartate 173 in catalysis. Cumulatively, this work... More