Biosynthesis and characterization of novel polyhydroxyalkanoate copolymers consisting of 3-hydroxy-2-methylbutyrate and 3-hydroxyhexanoate

Abstract

In this study, in order to explore the possibility of biosynthesizing a novel polyhydroxyalkanoate (PHA), copolymerization of 3-hydroxy-2-methylbutyrate (3H2MB) as the α-position methylated monomer and 3-hydroxyhexanoate (3HHx) as the medium-chain-length monomer was performed to obtain P(3H2MB-co-3HHx). The β-oxidation-deficient Escherichia coli LSBJ, harboring the PHA biosynthetic operon from Aeromonas caviae and the propionyl-CoA transferase gene (pct) from Megasphaera elsdenii, was cultured with feeding tiglic and hexanoic acids as the precursors for 3H2MB and 3HHx, respectively. It was observed that pct expression was highly effective to enhance the incorporation of 3H2MB into PHA. The biosynthesized PHA was composed of 3H2MB and 3HHx units only, and the 3H2MB fraction varied in the range of 36–60 mol% depending on the culture conditions. These PHAs exhibited glass transition temperatures between −11 to −17 °C; moreover, no melting peak was observed during analysis using differential scanning calorimetry. This study demonstrated the biosynthesis of a hitherto unreported PHA by engineering metabolic pathway in E. coli.