The impact of biomass withdrawal strategy on the biomass selection and polyhydroxyalkanoates accumulation of mixed microbial cultures.

The production of polyhydroxyalkanoates (PHA) by mixed microbial cultures (MMC) has been studied as an alternative to pure cultures in order to reduce the price of PHA through use of open systems and low-cost substrates, such as agro-industrial sub-products. However, the widespread applicability of this process depends on the optimization of operational factors impacting PHA productivity. This study addresses the impact of biomass withdrawal strategy on the performance of MMC selection reactors and consequently on biomass productivity and global PHA productivity. Two selection reactors were operated in parallel under similar conditions, except for the timing of biomass withdrawal, at the end of the famine phase (Reactor 1, R1) versus at the end of the feast phase (Reactor 2, R2) at an organic loading rate of 100 Cmmol.L-1.d-1 and solids retention time of 4 days. The biomass selected in both conditions had similar PHA storing capacity as shown by the similar yields of PHA per substrate obtained in the accumulation assays; however, R1 reached a higher biomass productivity (about 4-fold higher than R2). This study demonstrated that removing the excess biomass at the end of the famine phase resulted in a much higher global PHA productivity and that the key parameter affecting the global PHA productivity of the 2-stage system was the volumetric biomass productivity. Results obtained provide important insight into how MMC systems can be best operated to maximize PHA productivity.

Sludge retention time impacts on polyhydroxyalkanoate productivity in uncoupled storage/growth processes.

The process involving mixed microbial cultures (MMCs) and waste-based substrates emerged as an alternative solution to reduce the market price of polyhydroxyalkanoates (PHAs). The selection of an efficient MMC that displays a significant PHA accumulation potential and a high growth rate is considered a key factor for the MMC PHA production feasibility. This study used a pilot plant to investigate the dynamics of growth vs storage in a mixed culture fed with fermented fruit waste under uncoupled carbon and nitrogen feeding. Varying sludge retention times (SRTs) (2 and 4 d) and organic loading rates (OLRs) (from 2.6 to 14.5 gCOD.L-1.d-1) were imposed for this purpose. Results showed that, regardless of the OLR imposed, cultures selected at lower SRT grew faster and more efficiently using stored PHA. However, they had inferior specific storage rates and accumulation capacity, resulting in lower PHA productivity. Additionally, the polymer storage yield was independent of the SRT, and was directly linked with the abundance of putative PHA-storers in the MMC. The high PHA productivity (4.6 ± 0.3 g.L-1.d-1) obtained for the culture selected at 4 d of SRT was 80% above that obtained for the lower SRT tested, underlining the importance of achieving a good balance between culture growth and accumulation capacity to increase the viability of the PHA-producing process from wastes.

Single-Step Purification of Ovalbumin from Egg White Using Aqueous Biphasic Systems.

The ability of aqueous biphasic systems (ABS) composed of polyethylene glycols of different molecular weights (PEG 400, 600 and 1000) and buffered aqueous solutions of potassium citrate/citric acid (pH = 5.0 - 8.0) to selectively extract ovalbumin from egg white was here investigated. Phase diagrams, tie-lines and tie-line lengths were determined at 25ºC and the partitioning of ovalbumin in these systems was then evaluated. Aiming at optimizing the selective extraction of ovalbumin in the studied ABS, factors such as pH, PEG molecular weight and amount of the phase-forming components were initially investigated with pure commercial ovalbumin. In almost all ABS, it was observed a preferential partitioning of ovalbumin to the polymer-rich phase, with extraction efficiencies higher than 90%. The best ABS were then applied in the purification of ovalbumin from the real egg white matrix. In order to ascertain on the ovalbumin purity and yield, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion high performance liquid chromatography (SE-HPLC) analyses were conducted, confirming that the isolation/purification of ovalbumin from egg white was completely achieved in a single-step with a recovery yield of 65%. The results obtained show that polymer-salt-based ABS allow the selective extraction of ovalbumin from egg white with a simpler approach and better performance than previously reported. Finally, it is shown that ovalbumin can be completely recovered from the PEG-rich phase by an induced precipitation using an inexpensive and sustainable separation platform which can be easily applied on an industrial scale.