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. 2017 Jan 13;292(8):3517–3530. doi: 10.1074/jbc.M116.762666

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Genetic verification of the TAA biosynthetic gene cluster. A, HPLC analysis of TAA in the culture supernatants of BMB2451 (pink line), BMB2451-ΔtbrA (dark green line), BMB2451-ΔtbrB (blue line), BMB2451-ΔtbrA:tbrA (red line), and BMB2451-ΔtbrB:tbrB (black line) recombinant strains. TAA standard (purple line) and the supernatant extract of BMB171 with empty vector pHT304 (light green line) were used as positive and negative controls, respectively. B, Q-TOF-MS confirmation of TAA production by heterologous expression of the tbr operon in BMB2451. C, Q-TOF-MS analysis of TAA presence in intracellular and extracellular fractions of BMB2451-ΔtbrA and BMB2451-ΔtbrB mutants. The m/z 173.0090 ion indicating the mass of [M − H]⁻ patterned aconitic acid was used to extract a TAA signal from the total ion chromatograms (TIC). Retention times of extracted CAA (green peak) and TAA (pink peak) signals are provided. AU, absorbance units.