Impact of phosphate limitation on PHA production in a feast-famine process.

Double-limitation systems have shown to induce polyhydroxyalkanoates (PHA) production in chemostat studies limited in e.g. carbon and phosphate. In this work the impact of double substrate limitation on the enrichment of a PHA producing community was studied in a sequencing batch process. Enrichments at different C/P concentration ratios in the influent were established and the effect on the PHA production capacity and the enrichment community structure was investigated. Experimental results demonstrated that when a double substrate limitation is imposed at a C/P ratio in the influent in a range of 150 (C-mol/mol), the P-content of the biomass and the specific substrate uptake rates decreased. Nonetheless, the PHA storage capacity remained high (with a maximum of 84 wt%). At a C/P ratio of 300, competition in the microbial community is based on phosphate uptake, and the PHA production capacity is lost. Biomass specific substrate uptake rates are a linear function of the cellular P-content, offering advantages for scaling-up the PHA production process due to lower oxygen requirements.

Survival of the fastest: Selective removal of the side population for enhanced PHA production in a mixed substrate enrichment.

The success of enriching PHA-producers in a feast/famine regime strongly depends on the substrate utilized. A distinction can be made between substrates that select for PHA-producers (e.g. volatile fatty acids) and substrates that select for growing organisms (e.g. methanol). In this study the feasibility of using such a mixed substrate was evaluated. A sedimentation step was introduced in the cycle after acetate depletion and the supernatant containing methanol was discharged. This process configuration resulted in an increased maximum PHA storage capacity of the biomass from 48wt% to 70wt%. A model based on the experimental results indicated that the length of the pre-settling period and the supernatant volume that is discharged play a significant role for the elimination of the side population. However, the kinetic properties of the two different populations determine the success of the proposed strategy.

PHA production from the organic fraction of municipal solid waste (OFMSW): Overcoming the inhibitory matrix.

Leachate from the source separated organic fraction of municipal solid waste (OFMSW) was evaluated as a substrate for polyhydroxyalkanoates (PHA) production. Initially, the enrichment step was conducted directly on leachate in a feast-famine regime. Maximization of the cellular PHA content of the enriched biomass yielded to low PHA content (29 wt%), suggesting that the selection for PHA-producers was unsuccessful. When the substrate for the enrichment was switched to a synthetic volatile fatty acid (VFA) mixture -resembling the VFA carbon composition of the leachate-the PHA-producers gained the competitive advantage and dominated. Subsequent accumulation with leachate in nutrient excess conditions resulted in a maximum PHA content of 78 wt%. Based on the experimental results, enriching a PHA-producing community in a "clean" VFA stream, and then accumulating PHA from a stream that does not allow for enrichment but does enable a high cellular PHA content, such as OFMSW leachate, makes the overall process much more economically attractive. The estimated overall process yield can be increased four-fold, in comparison to direct use of the complex matrix for both enrichment and accumulation.

Biosurfactant production from marine hydrocarbon-degrading consortia and pure bacterial strains using crude oil as carbon source.

Biosurfactants (BSs) are "green" amphiphilic molecules produced by microorganisms during biodegradation, increasing the bioavailability of organic pollutants. In this work, the BS production yield of marine hydrocarbon degraders isolated from Elefsina bay in Eastern Mediterranean Sea has been investigated. The drop collapse test was used as a preliminary screening test to confirm BS producing strains or mixed consortia. The community structure of the best consortia based on the drop collapse test was determined by 16S-rDNA pyrotag screening. Subsequently, the effect of incubation time, temperature, substrate and supplementation with inorganic nutrients, on BS production, was examined. Two types of BS - lipid mixtures were extracted from the culture broth; the low molecular weight BS Rhamnolipids and Sophorolipids. Crude extracts were purified by silica gel column chromatography and then identified by thin layer chromatography and Fourier transform infrared spectroscopy. Results indicate that BS production yield remains constant and low while it is independent of the total culture biomass, carbon source, and temperature. A constant BS concentration in a culture broth with continuous degradation of crude oil (CO) implies that the BS producing microbes generate no more than the required amount of BSs that enables biodegradation of the CO. Isolated pure strains were found to have higher specific production yields than the complex microbial marine community-consortia. The heavy oil fraction of CO has emerged as a promising substrate for BS production (by marine BS producers) with fewer impurities in the final product. Furthermore, a particular strain isolated from sediments, Paracoccus marcusii, may be an optimal choice for bioremediation purposes as its biomass remains trapped in the hydrocarbon phase, not suffering from potential dilution effects by sea currents.