Microbial polyhydroxyalkanoates (PHAs) are promising biodegradable polymers that may alleviate some of the environmental burden of petroleum-derived polymers. The requirements for carbon substrates and energy for bioreactor operations are major factors contributing to the high production costs and environmental impact of PHAs. Improving the process productivity is an important aspect of cost reduction, which has been attempted using a variety of fed-batch, continuous, and semi-continuous bioreactor systems, with variable results. The purpose of this review is to summarize the bioreactor operations targeting high PHA productivity using pure cultures. The highest volumetric PHA productivity was reported more than 20 years ago for poly(3-hydroxybutryate) (PHB) production from sucrose (5.1 g L⁻¹ h⁻¹). In the time since, similar results have not been achieved on a scale of more than 100 L. More recently, a number fed-batch and semi-continuous (cyclic) bioreactor operation strategies have reported reasonably high productivities (1 g L⁻¹ h⁻¹ to 2 g L⁻¹ h⁻¹) under more realistic conditions for pilot or industrial-scale production, including the utilization of lower-cost waste carbon substrates and atmospheric air as the aeration medium, as well as cultivation under non-sterile conditions. Little development has occurred in the area of fully continuously fed bioreactor systems over the last eight years.

Glucose and xylose are fermentable sugars readily available from lignocellulosic biomass, and are a sustainable carbon substrate supporting industrial biotechnology. Three strains were assessed in this work - Paraburkholderia sacchari, Hydrogenophaga pseudoflava, and Bacillus megaterium - for their ability to uptake both C₅ and C₆ sugars contained in a hardwood hydrolysate produced via a thermomechanical pulping-based process with concomitant production of poly(3-hydroxyalkanoate) (PHA) biopolymers. In batch conditions, B. megaterium showed poor growth after 12 h, minimal uptake of xylose throughout the cultivation, and accumulated a maximum of only 25 % of the dry biomass as PHA. The other strains simultaneously utilized both sugars, although glucose uptake was faster than xylose. From hardwood hydrolysate, P. sacchari accumulated 57 % of its biomass as PHA within 24 h, whereas H. pseudoflava achieved an intracellular PHA content of 84 % by 72 h. The molecular weight of the PHA synthesized by H. pseudoflava (520.2 kDa) was higher than that of P. sacchari (265.5 kDa). When the medium was supplemented with propionic acid, the latter was rapidly consumed by both strains and incorporated as 3-hydroxyvalerate subunits into the polymer, demonstrating the potential for production of polymers with improved properties and value. H. pseudoflava incorporated 3-hydroxyvalerate subunits with at least a 3-fold higher yield, and produced polymers with higher 3-hydroxyvalerate content than P. sacchari. Overall, this work has shown that H. pseudoflava can be an excellent candidate for bioconversion of lignocellulosic sugars to PHA polymers or copolymers as part of an integrated biorefinery.

The rheology of high-cell density (HCD) cultures is an important parameter for its impact on mixing and sparging, process scale-up, and downstream unit operations in bioprocess development. In this work, time-dependent rheological properties of HCD Pseudomonas putida LS46 cultures were monitored for microbial polyhydroxyalkanoate (PHA) production. As the cell density of the fed-batch cultivation increased (0 to 25 g·L⁻¹ cell dry mass, CDM), the apparent viscosity increased nearly nine-fold throughout the fed-batch process. The medium behaved as a nearly Newtonian fluid at lower cell densities, and became increasingly shear-thinning as the cell density increased. However, shear-thickening behavior was observed at shearing rates of approximately 75 rad·s⁻¹ or higher, and its onset increased with viscosity of the sample. The supernatant, which contained up to 9 g·L⁻¹ soluble organic material, contributed more to the observed viscosity effect than did the presence of cells. Owing to this behavior, the oxygen transfer performance of the bioreactor, for otherwise constant operating conditions, was reduced by 50% over the cultivation time. This study has shown that the dynamic rheology of HCD cultures is an important engineering parameter that may impact the final outcome in PHA cultivations. Understanding and anticipating this behavior and its biochemical origins could be important for improving overall productivity, yield, process scalability, and the efficacy of downstream processing unit operations.

Medium chain‐length polyhydroxyalkanoates (mcl‐PHAs) are efficiently synthesized from fatty acids. It was hypothesized that under non‐axenic conditions, the anti‐microbial properties of fatty acids would reduce growth of microbial competitors and allow Pseudomonas putida LS46 to dominate the population resulting in non‐sterile mcl‐PHA production. Non‐sterile batch and fed‐batch cultures produced similar final mcl‐PHA content, monomer composition, and productivity during 24–27 h cultivations when compared to sterile control cultures for initial octanoic acid concentrations ranging from 5 to 40 mM at an initial pH of 6.5. In the absence of a P. putida inoculum, growth was eventually observed in non‐sterile medium after a lag period of up to 120 h, the length of which was dependent on the octanoic acid concentration. The efficacy of prolonged non‐sterile cultivation was tested using a sequential batch reactor (SBR). After 28 days (672 h), 1.62 g L⁻¹ of total biomass was observed, of which the mcl‐PHA content was 47%. This resulted in a PHA titer of 0.77 g L⁻¹, and a volumetric PHA productivity of 32 mg L⁻¹ h⁻¹. The polymer composition remained stable throughout at 87.0 ± 2.7% C₈ monomers, and short‐chain length (scl‐) monomers were not detected. This study is the first of its kind to report that purely mcl‐PHAs were produced after prolonged periods in a non‐sterile environment and demonstrate that medium chain‐length fatty acids exert a strong selective pressure toward organisms that synthesize mcl‐PHA. This suggests an opportunity for mcl‐PHA production in open continuous cultivation, which could reduce both fixed and operating costs.