Fig 5.

HPLC chromatograms of CoA activation assays with cell extracts of Pseudomonas sp. strain Chol1 containing DHOPDC (XII in Fig. 1) as the substrate. (A) Representative t₀ for all assays. (B) Assay with nondesalted cell extracts incubated for 60 and 120 min. (C) Assay with desalted cell extracts incubated for 120 min. (D) Assay with desalted cell extracts in the presence of NAD⁺ incubated for 120 min. (E) Assay with desalted cell extracts in the presence of NAD⁺ and PMS incubated for 120 min. P3 was identified as the CoA ester of DHOPDC (XIII), P4 was identified as 12α-DHADD (XVI), and P6 was identified as HADT (XVII). The analysis wavelengths were 245 nm (black) and 260 nm (gray).