Production and characterization of ultra-high-molecular weight poly(3-hydroxybutyrate) by recombinant Escherichia coli

An efficient fermentation strategy for the high level production of ultra-high-molecular weight poly(3-hdyroxybutyrate) (PHB) was developed. Although the cell and PHA concentrations obtained by flask cultures at different initial pH (6.0 or 6.9) were almost same level, the molecular mass of PHB produced were quite different along with the initial pH. When a recombinant Escherichia coli XL1-Blue harboring pJC2 containing the Alcaligenes latus PHB biosynthesis genes was cultivated in flask culture (pH 6.0), the PHB having a very high molecular weight of 22 MDa could be produced while only below 1 MDa at initial pH 6.9. The ultra-high-molecular weight PHB could be synthesized to high concentration of 89.8 g/L resulting in the PHB productivity of 2.07 g/L-h by simple fed-batch culture. In this study, we report that PHB having various molecular mass can be produced by employing metabolically engineered E. coli strains harboring the plasmids of different copy numbers containing the A. latus phbCAB genes.