Characterization of YbjG, a pyrophosphate phosphatase from E. coli involved in the lipid carrier undecaprenyl phosphate metabolism

•Background
Undecaprenyl phosphate (C55-P) is an essential lipid carrier involved in the biosynthesis of cell surface carbohydrate polymers such as the peptidoglycan. C55-P is the result of the dephosphorylation of the undecaprenyl pyrophosphate (C55-PP) by specific phosphatases. In Escherichia coli this dephosphorylation can be performed by four integral membrane proteins, BacA, and three members of the type 2 phosphatidic acid phosphatase family (PAP2), PgpB, YbjG, and LpxT.
•Objectives
The aim of this project is to characterize YbjG and contributes to the understanding of the physiological role and mechanism of action of this enzyme in the C55-P metabolism. The C55-PP phosphatases could become an interesting target in the search for new molecules with antibacterial activity.
•Methods
In parallel the stability of YbjG and its activity against C15-PP were assessed in 94 different detergents. Moreover the enzymatic activity of YbjG was studied: substrate specificity, optimum pH and temperature, effect of detergent concentration.
•Conclusions
For the first time, YbjG has been purified and we show its ability to dephosphorylate C15-PP, DGPP and C55-PP in vitro with respectively decreasing efficiency. No activity has been detected on five other potential substrates (PPi, PA, C5-PP, G6P & PNPP).
The phosphatase activity on C15-PP is maximum at pH 6,5 and 25 °C. Moreover Cymal6, LMNG, & ωUDM are good detergent both for the stability and the C15-PP phosphatase activity of YbjG, but approximately half of the 94 tested detergents show C15-PP phosphatase activity on the qualitative enzymatic test.