In Vitro Reconstitution of Two Essential Steps in Wall Teichoic Acid Biosynthesis

ACS Chemical Biology (Impact Factor: 5.33). 03/2006; 1(1):25-8. DOI: 10.1021/cb0500041
Source: PubMed


Wall teichoic acids (WTAs) are anionic polymers that decorate the cell walls of many gram-positive bacteria. These structures are essential for survival or virulence in many organisms, which makes the enzymes involved in their biosynthesis attractive targets for the development of new antibacterial agents. We present a strategy to obtain WTA biosynthetic intermediates that involves a combination of chemical and enzymatic transformations. Using these intermediates, we have reconstituted the first two committed steps in the biosynthetic pathway. This work enables the exploration of WTA-synthesizing enzymes as antibiotic targets.

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    • "Furthermore , the TagA reaction product was used as a substrate for recombinantly purified TagB, verifying the glycerophosphotransferase activity previously reported (Bhavsar et al., 2005). The use of a soluble acceptor in high concentration translated into a reaction turnover number that was vastly higher to that reported with endogenous substrates (Bhavsar et al., 2005; Ginsberg et al., 2006) and holds tremendous potential for the detailed characterization of enzymes that can utilize this undecaprenol analogue, e.g. TagO, TagA, TagB and TagF. "
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