Effect of complex nitrogen source on the synthesis and accumulation of poly(3-hydroxybutyric acid) by recombinantEscherichia coli in flask and fed-batch cultures

Abstract

When a recombinantEscherichia coli XL1-Blue harboring pSYL105 was cultured in a complex medium, a poly(3-hydroxybutyric acid) (PHB) concentration of 7.16 g/L was obtained in 48 h. However, a PHB concentration of only 0.91 g/L was obtained in 60 h by culturing in a defined medium. Also, fed-batch culture in a defined medium resulted in considerably lower PHB accumulation than in a complex medium. With the aim to produce a high concentration of PHB at a reduced medium cost, we examined 10 complex nitrogen sources for their ability to promote PHB synthesis in a defined medium. Tryptone, casamino acids, and casein hydrolysate promoted PHB synthesis to a higher extent than the others tested. PHB synthesis was also enhanced during fedbatch cultures when a defined medium was supplemented with various complex nitrogen sources. With tryptone supplementation a PHB concentration of 66.7 g/L could be obtained in 44 h. Yeast extract was less effective for promoting PHB synthesis than tryptone. Corn steep liquor, which did not enhance PHB synthesis significantly, could promote PHB synthesis considerably when supplemented together with yeast extract in both flask and fed-batch cultures.