Fig. 6.

Preparation of soluble monooxygenases. a Group 2 enzymes catalyze an oxidization reaction of phytosterol at the C26 position. b PCR amplification of the Mon164 gene from the HGMS2 genome indicated the expected sizes. The arrow indicates the expected PCR band. c PCR amplification of the Mon197 gene from the HGMS2 genome with a gradient annealing temperature. The arrow indicates the expected PCR band. The DNA marker was run on the same gel in c. d Mon164 was overexpressed in soluble form and fused with a His-tag. S, supernatant; P, pellets and M, molecular weight marker. e Mon197 was overexpressed in soluble form in fusion with a His6-MBP-tag. S, supernatant; P, pellet and M, molecular weight marker