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• Supplemental file 1 - Descriptions of constructs and cloning vectors, construction of pECO-1 vector, cloning of heterodimer of liver X receptors and elongin C, and cloning of P. aeruginosa pgl; culture conditions for high-cell-density fed-batch cultures of E. coli growing in a completely defined medium; details of analytical methods to measure in vitro PGL activity, including the correlation of maximum PGL activity vs adduct formation in fed-batch cultures of E. coli BL21(DE3) (Table S1); HPLC assay for 18-kDa protein, LC/MS assay for heterologous proteins, LC/MS/MS analysis of gluconoylated 18-kDa protein; heterologous protein preparation of elongin C and heterodimer of liver X receptors; determination of E. coli macromolecular composition required for metabolic flux analysis (Table S2); nucleotide sequence for the cloning vector pECO-1 (Fig. S1); restriction enzyme map of the final 3,416-bp construct, pECO-1-pgl (Fig. S2); detection of the 18-kDa gluconoylation product by MS (Fig. S3); and identification of gluconoylated amino acids in the 18-kDa protein sequence by LC/MS/MS (Fig. S4).
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