Cell-recycle fed-batch production of a highly unsaturated polyhydroxyalkanoate from 1,3-butanediol byPseudomonas sp. A33

The recently isolatedPseudomonas sp. A33 was investigated for the production of a highly unsaturated polyhydroxyalkanoate (PHA) containing various alkyl and alkenyl pendent groups from 1,3-butanediol in a cell-recycle fed-batch production mode. The monomer composition and degree of unsaturation in PHA were dependent on the environmental conditions. The production temperature markedly influenced the content, composition, and degree of unsaturation of PHA. As the production temperature decreased from 30 to 10°C, the degree of unsaturation and content of PHA were increased, while the mole percentage of 3-hydroxybutyrate (3HB) was decreased. These temperature effects on the composition of PHA imply that the production can be used as a control variable for the biosynthesis of a highly unsaturated PHA and for the specific regulation of the composition of PHA. The biosynthetic pathway for a highly unsaturated PHA which is based on de novo fatty acid biosynthetic pathway is proposed. For the enhanced production of this functional PHA, a high cell density was achieved by cell-recycle continuous culture at 30°C, and then a large amount of PHA was accumulated at 15°C by fed-batch addition of the feeding solution containing excess 1,3-butanediol. The structures of monomer constituents of polymer were confirmed by gas chromatography—mass spectrometric analysis of trimethylsiyl derivatives of 3-hydroxyalkanoic acids methyl esters.