Enhanced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production from volatile fatty acids by Halomonas sp. YJ01 with 2-methylcitrate cycle

Volatile fatty acids (VFAs) are suitable substrates for synthesizing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), wherein propionate is a precursor of PHBV biosynthesis; however, high concentrations are toxic to bacteria. Therefore, VFAs with suitable ratio are needed. Here, with the ratio of acetate: propionate: butyrate being 1:4:2, the maximum PHBV content and the 3HV content were 46.77 wt% and 19.24 mol%, respectively, by Halomonas sp. YJ01. The optimal C/P and C/N ratios for PHBV synthesis were controlled at 800-1000 and 70-90. The carbon source uptake by the strain at higher C/N ratios was mainly used to synthesize PHBV. The metabolic pathway for PHBV biosynthesis with mixed VFAs showed that the 2-methylcitrate cycle (2-MCC) pathway converted propionyl-CoA to pyruvate, which reduced the toxicity of propionate to the strain. Moreover, the strain utilized acetate and butyrate without producing pyruvate, which did not affect the detoxification of the 2-MCC pathway. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) results showed that when the 2-MCC pathway was inhibited, phaC expression decreased 2.74-fold, and the expression of prpB and prpC was down-regulated 2-fold and 6.88-fold, respectively; therefore, propionate toxicity exposure resulted in a significant decrease in PHBV content.