Production and characterization of homopolymer polyhydroxyheptanoate (P3HHp) by a fadBA knockout mutant Pseudomonas putida KTOY06 derived from P. putida KT2442

Abstract

Pseudomonas putida KT2442 commonly produces medium-chain-length polyhydroxyalkanoates (PHA) consisting of 3-hydroxyhexanoate (C6) to 3-hydroxydodecanoate (C12) when grown in glucose or even number fatty acid. When two of the beta-oxidation genes fadBA were deleted, the P. pudida KT2442 mutant named KTOY06 accumulated a homopolymer of poly-3-hydroxyheptanoate (P3HHp) up to 71 wt% of its cell dry weight in the presence of heptanoate as a single carbon source. P3HHp contents in the cell dry weight were in direct proportional to Na-heptanoate concentration up to 10 g/L. In contrast, under the same cultivation conditions, the wild type P. putida KT2442 produced a copolymer consisting of 3-hydroxyheptanoate (3HHp) and 5.3–8.4 mol% 3-hydroxynonanoate (3HN). Gas chromatography (GC), nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and gel permeation chromatography (GPC) were used to characterize the homopolymer P3HHp, respectively. It was found that the P3HHp with an average molecular weight of 455 kDa was a completely amorphous homopolymer without crystallinity. P3HHp is thermo-degradable at around 250 °C.