Methanotrophic Poly(hydroxybutyrate) Through C1 Fermentation and Downstream Process Development: Molar Mass, Thermal and Mechanical Characterization

Today, PHB and its copolymers-potential plastic substitutes-are produced by fermenting sugar, which is not scalable to the volumes of plastic consumption. PHB from CH₄ can offer a sustainable process route, with CH₄ potentially produced from a variety of waste biomass streams through anaerobic digestion, gasification, and methanation. The high molar mass (M_w) of PHB is a key determinant of its mechanical properties, and strain, culture conditions and downstream processing influence it. In this work, the strain <i>Methylocystis</i> sp. GB 25 (DSMZ 7674) was grown on natural gas as the sole carbon and energy source and air (1:1) in a loop reactor with 350 L active fermentation volume, at 35 °C and ambient pressure. After two days of continuous growth, the bacteria were limited in P and N for 1, 2, and 2.5 days to determine the optimal conditions for PHB accumulation and the highest Mw as the target. The biomass was then centrifuged and spray-dried. For downstream processing, chloroform solvent extraction and selected enzymatic treatment were deployed, yielding ~40% PHB from the biomass. The PHB obtained by solvent extraction exhibited high average weight molar masses of <i>M</i>_w ~1.1-1.5 × 10⁶ g mol^-1. The highest <i>M</i>_w was obtained after one day of limitation, whereas enzyme treatment resulted in partially degraded PHB. Cold chloroform maceration, interesting due to energy savings, did not achieve sufficient extraction efficiency because it was unable to extract high-molar-mass PHB fractions. The extracted PHB has a high molar mass, more than double that of standard commercial PHB, and was characterized by DSC, which showed a high degree of crystallinity of up to 70% with a melting temperature of close to 180 °C. Mechanical tensile properties measurements, as well as dynamic mechanical thermal analysis (DMTA), were performed. Degradation of the PHB by enzymes was also determined. Methanotrophic PHB is a promising bioplastics material. The high M_w can limit and delay polymer degradation in practical processing steps, making the material more versatile and robust.