Biosynthesis of Polyhydroxyalkanoate Terpolymer from Methanol via the Reverse β-Oxidation Pathway in the Presence of Lanthanide

AM1 is the attractive platform for the production of value-added products from methanol. We previously demonstrated that equipped with PHA synthase with broad substrate specificity synthesized polyhydroxyalkanoates (PHAs) composed of ( )-3-hydroxybutyrate and small fraction of ( )-3-hydroxyvalerate (3HV) and ( )-3-hydroxyhexanoate (3HHx) units on methanol. This study further engineered for biosynthesis of PHAs with higher 3HV and 3HHx composition focusing on the EMC pathway involved in C assimilation. The introduction of ethylmalonyl-CoA decarboxylase, catalyzing a backward reaction in the EMC pathway, aiming to increase intracellular propionyl/butyryl-CoA precursors did not affect PHA composition. Reverse β-oxidation pathway and subsequent ( )-specific hydration of 2-enoyl-CoA were then enhanced by heterologous expression of four genes derived from for the conversion of propionyl/butyryl-CoAs to the corresponding ( )-3-hydroxyacyl-CoA monomers. The resulting strains produced PHAs with higher 3HV and 3HHx compositions, while the methylotrophic growth was severely impaired. This growth impairment was interestingly restored by the addition of La without a negative impact on PHA biosynthesis, suggesting the activation of the EMC pathway by La . The engineered synthesized PHA terpolymer composed of 5.4 mol% 3HV and 0.9% of 3HHx with 41% content from methanol as a sole carbon source in the presence of La .