Fig 8.

Chromatograms of enzyme assays with desalted cell extracts of Pseudomonas sp. strain Chol1 containing P6 as the substrate. (A) Representative t₀ for all assays. (B) Assay with NADH incubated for 30 min. (C) Assay with NADPH incubated for 30 min. P6 was identified as HADT (XVII in Fig. 1). The analysis wavelength was 245 nm. (D) Isomerization of 12α-DHADD to 12β-DHADD via HADT. The oxidation to HADT is NAD⁺ dependent, and the back reaction could be measured in vitro; the reduction of HADT to 12β-DHADD is NADPH dependent; 12β-DHADD is further degraded to THSATD.