2.7. Confirmation of the proposed dihydroxylation pathway
To further confirm the proposed dihydroxylation pathway,
biotransformations of intermediate products purified from transformation
extract with C. lini ST-1 were performed. After C. lini ST-1
was cultivated in fermentation medium for 24 h under the same
conditions as described in Section 2.3, the fine powder of purified
products (2 g/L) was added into the medium. Transformation
reaction was carried out in 250mL flasks containing 30mL of fermentation
medium on a rotary shaker (220 rpm) at 28 ◦C for 72 h.
Samples were withdrawn at regular intervals and transformation
products were subsequently extracted with ethyl acetate for three
times and dried in vacuum. The analysis of product concentrations
were the same as described in Section 2.6.