Koh, Francis H.

The fatty acid synthetase (FAS) from the marine invertebrate Bugula neritina was isolated using cold ethanol precipitation followed by the sequence of G-100 and G-200 size exclusion columns. Native gel analysis indicates the isolation of the FAS and the elution volume from the G-100 column suggests the FAS to be ~282kd. One band with a molecular weight of 66 kDa appeared on the SDS gel of the G-200 sample (F17-30) that eluted at 52.5 mL. The G-200 sample that eluted at 19.2 mL (F1-6) displayed two predominant bands on the SDS gel corresponding to molecular weights 66 kDa and 97 kDa. Both F1-6 and F17-30 showed FAS activity displaying de novo production of myristic and palmitic acids. From the sequence of purification starting from the cell-free extract (CFE) to the F17-30 sample, a 240 fold increase in specific activity was observed. The Type II FAS experiments showed no substantial evidence of activity, namely of the beta-Hydroxybutyryl acyl-dehydrase and the enoyl reductase enzymes.