Polyhydroxyalkanoates (PHAs) are biobased and biodegradable polymers that may be considered to replace fossil-based materials. However, their widespread deployment is slowed down because of the high production cost and their low quality. Using methane during the generation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-co-HV) could reduce the production costs, which are usually owed to the carbon source used, and enhance the performance of the material. In fact, the properties of PHB-co-HV make it more suitable than the most common homopolymer, named poly(3-hydroxybutyrate) (PHB), for many applications since it is more flexible, has a wider thermal processing range, a lower crystallinity and permeability to water. In this work, the production of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) was simulated using an innovative process scheme consisting of two bioreactors working in series equipped with a gas recycling unit, which was designed to increase the methane mass transfer from the gas to the liquid. During the first step, a Methylocystis dominated culture was grown on ammonia and methane to reach a high cell concentration; then, the cells were moved to the second bioreactor and subjected to nutrients starvation to favour the metabolic pathway addressed to the accumulation of PHB-co-HV. The effect of the variation of valeric acid concentration (100-400 ppm), which was used as co-substrate, on the fraction of PHB-co-HV stored in the cells and the percentage of 3-hydroxyvalerate (3-HV) was investigated: results showed that the highest the valeric acid concentration, the lower the total polymer content and the higher the HV fraction accumulated.