Search Results (14)

Food industry waste and wastewater have been explored in relation to acidogenic fermentation as sources of non-competing food carbohydrates and mixed microbial cultures (MMCs), respectively, with the aim of producing short-chain organic acids (SCOAs) with general applications in polyhydroxyalkanoates (PHAs) production. However, studies on acidogenic fermentation using lipidic substrates are scarce. In this work, it was hypothesized that spent frying oil (SFO) could be used as a substrate for SCOA production via MMCs. In this study, oleic acid was used as a model molecule. The characterization of SFO revealed that it is mainly composed of oleic acid (81%), with minor amounts of palmitic, linoleic, and stearic acids. Different MMCs and food-to-microorganism (F/M) ratios were tested. MMCs collected in the aerobic tank of a municipal wastewater treatment plant (AES), at a 1:1 F/M, allowed to obtain the highest SCOA concentration (1.50 g COD/L) and the most diverse profile of SCOAs, with the production of acetic, propionic, butyric, iso-butyric, and valeric acids at 48:17:9:13:13% on a molar basis, respectively. This variety of odd and even SCOAs is of upmost importance, with potential applications in producing PHAs. This work can be considered a starting point for future acidogenic fermentation studies using lipid-based substrates and for the future production of PHAs. Full article

Raw brewers’ spent grain (BSG), a by-product of beer production and produced at a large scale, presents a composition that has been shown to have potential as feedstock for several biological processes, such as polyhydroxyalkanoates (PHAs) production. Although the high interest in the PHA production from waste, the bioconversion of BSG into PHA using microbial mixed cultures (MMC) has not yet been explored. This study explored the feasibility to produce PHA from BSG through the enrichment of a mixed microbial culture in PHA-storing organisms. The increase in organic loading rate (OLR) was shown to have only a slight influence on the process performance, although a high selectivity in PHA-storing microorganisms accumulation was reached. The culture was enriched on various PHA-storing microorganisms, such as bacteria belonging to the Meganema, Carnobacterium, Leucobacter, and Paracocccus genera. The enrichment process led to specialization of the microbiome, but the high diversity in PHA-storing microorganisms could have contributed to the process stability and efficiency, allowing for achieving a maximum PHA content of 35.2 ± 5.5 wt.% (VSS basis) and a yield of 0.61 ± 0.09 Cmmol_PHA/Cmmol_VFA in the accumulation assays. Overall, the production of PHA from fermented BSG is a feasible process confirming the valorization potential of the feedstock through the production of added-value products. Full article

The use of fossil-based plastics has become unsustainable because of the polluting production processes, difficulties for waste management sectors, and high environmental impact. Polyhydroxyalkanoates (PHA) are bio-based biodegradable polymers derived from renewable resources and synthesized by bacteria as intracellular energy and carbon storage materials under nutrients or oxygen limitation and through the optimization of cultivation conditions with both pure and mixed culture systems. The PHA properties are affected by the same principles of oil-derived polyolefins, with a broad range of compositions, due to the incorporation of different monomers into the polymer matrix. As a consequence, the properties of such materials are represented by a broad range depending on tunable PHA composition. Producing waste-derived PHA is technically feasible with mixed microbial cultures (MMC), since no sterilization is required; this technology may represent a solution for waste treatment and valorization, and it has recently been developed at the pilot scale level with different process configurations where aerobic microorganisms are usually subjected to a dynamic feeding regime for their selection and to a high organic load for the intracellular accumulation of PHA. In this review, we report on studies on terrestrial and marine bacteria PHA-producers. The available knowledge on PHA production from the use of different kinds of organic wastes, and otherwise, petroleum-polluted natural matrices coupling bioremediation treatment has been explored. The advancements in these areas have been significant; they generally concern the terrestrial environment, where pilot and industrial processes are already established. Recently, marine bacteria have also offered interesting perspectives due to their advantageous effects on production practices, which they can relieve several constraints. Studies on the use of hydrocarbons as carbon sources offer evidence for the feasibility of the bioconversion of fossil-derived plastics into bioplastics. Full article

Two eco-friendly healing bioproducts generated from microbial mixed cultures (MMC) for the production of polyhydroxyalkanoates (PHA) were used as surface treatments, with two residual materials used as the substrates, namely crude glycerol and pinewood bio-oil. Their ability to improve the durability of concrete samples containing recycled aggregates was assessed. To determine this protective capacity, 180 samples were analyzed using different tests, such as water penetration under pressure, capillary absorption, freeze–thaw and water droplet absorption test. Three types of conditions were used: outdoor–indoor exposure, re-application of biopolymers and application in vertical exposure conditions. The results showed reductions of up to 50% in the water penetration test and a delay in the water droplet absorption test of up to 150 times relative to the reference. The surface application of these bioproducts significantly reduced the degree of water penetration in recycled concrete, increasing its useful lifespan and proving to be a promising treatment for protecting concrete surfaces. Full article

The fishing industry produces vast amounts of saline organic side streams that require adequate treatment and disposal. The bioconversion of saline resources into value-added products, such as biodegradable polyhydroxyalkanoates (PHAs), has not yet been fully explored. This study investigated PHA production by mixed microbial cultures under 30 g_NaCl/L, the highest NaCl concentration reported for the acclimatization of a PHA-accumulating mixed microbial culture (MMC). The operational conditions used during the culture-selection stage resulted in an enriched PHA-accumulating culture dominated by the Rhodobacteraceae family (95.2%) and capable of storing PHAs up to 84.1% wt. (volatile suspended solids (VSS) basis) for the highest organic loading rate (OLR) applied (120 Cmmol/(L.d)). This culture presented a higher preference for the consumption of valeric acid (0.23 ± 0.03 Cmol_HVal/(Cmol_X.h)), and the 3HV monomer polymerization (0.33 ± 0.04 Cmmol_HV/(Cmmol_X.h) was higher as well. As result, a P(3HB-co-3HV)) with high HV content (63% wt.) was produced in the accumulation tests conducted at higher OLRs and with 30 g_NaCl/L. A global volumetric PHA productivity of 0.77 gPHA/(L.h) and a specific PHA productivity of 0.21 gPHA/(gX.h) were achieved. These results suggested the significant potential of the bioconversion of saline resources into value-added products, such as PHAs. Full article

It is urgent to expand the market of biodegradable alternatives to oil-derived plastics owing to (i) increasingly limited oil availability/accessibility, and (ii) the dramatic impact of traditional plastics on aquatic life, the food chain, all Earth ecosystems, and ultimately, human health. Polyhydroxyalkanoates (PHAs) are promising biodegradable polymers that can be obtained through microbial fermentation of agro-industrial byproducts, e.g., milk and cheese whey. Here, the PHA-accumulating efficiency of a mixed microbial culture (MMC, derived from activated sludges) grown on dairy byproducts (cheese and scotta whey) was measured. Bioreactor tests featuring temperature and pH control showed that both scotta and pre-treated Toma cheese whey could be used for PHA production by MMC, although scotta cheese whey supported higher PHA yield and productivity. The advantages of open MMCs include their plasticity and versatility to fast changing conditions; furthermore, no growth-medium sterilization is needed prior to fermentation. However, the use of pure cultures of efficient PHA producers may support better metabolic performances. Therefore, PHA-producing strains were isolated from a MMC, leading to the satisfactory identification of two bacterial strains, Citrobacter freundii and Leuconostoc spp., whose ability to accumulate PHAs in synthetic media was confirmed. A more detailed investigation by mass spectrometry revealed that the strain was L. mesenteroides. Although the validation of L. mesenteroides potential to produce PHA through fermentation of agro-industrial byproducts requires further investigations, this is the first study reporting PHA production with the Leuconostoc genus. Full article

One approach to tackle the problems created by the vast amounts of construction and demolition waste (CDW) generated worldwide while at the same time lengthening concrete durability and service life is to foster the use of recycled aggregate (RA) rather than natural aggregate (NA). This article discusses the use of polyhydroxyalkanoates (PHAs)-producing mixed microbial cultures (MMCs) to treat the surface of recycled concrete with a view to increase its resistance to water-mediated deterioration. The microorganisms were cultured in a minimal medium using waste pinewood bio-oil as a carbon source. Post-application variations in substrate permeability were determined with the water drop absorption and penetration by water under pressure tests. The significant reduction in water absorption recorded reveals that this bioproduct is a promising surface treatment for recycled concrete. Full article

The wastewater from the cookers of a tuna-canning plant was used as feedstock for the process. It was acidified in a continuous stirred tank reactor (CSTR) of 1.5 L to produce a mixture of volatile fatty acids (VFAs). The effluent contained 28.3 ± 8.7 g COD_S/L and 25.0 ± 4.6 g COD_VFA/L, 4.4 ± 1.6 g NH₄⁺/L, and 10.9 ± 4.0 g Na⁺/L, which corresponds to about 28 g NaCl/L approximately. This was used to feed a PHA production system. The enriched MMC presented a capacity to accumulate PHAs from the fermented tuna wastewater. The maximum PHA content of the biomass in the fed-batch (8.35 wt% PHA) seemed very low, possibly due to the variable salinity (from 2.2 up to 12.3 g NaCl/L) and the presence of ammonium (which promoted the biomass growth). The batch assay showed a PHA accumulation of 5.70 wt% PHA, but this is a much better result if the productivity of the reactor is taken into account. The fed-batch reactor had a productivity of 10.3 mg PHA/(L h), while the batch value was about five times higher (55.4 mg PHA/(L h)). At the sight of the results, it can be seen that the acidification of fish-canning wastewater is possible even at high saline concentrations (27.7 g NaCl/L). On the other hand, the enrichment and accumulation results show us promising news and which direction has to be followed: PHAs can be obtained from challenging substrates, and the feeding mode during the accumulation stage has an important role to play when it comes to inhibition. Full article

The large increase in the world population has resulted in a very large amount of construction waste, as well as a large amount of waste glycerol from transesterification reactions of acyl glycerides from oils and fats, in particular from the production of biodiesel. Only a limited percentage of these two residues are recycled, which generates a large management problem worldwide. For that reason, in this study, we used crude glycerol as a carbon source to cultivate polyhydroxyalkanoates (PHA)-producing mixed microbial cultures (MMC). Two bioproducts derived from these cultures were applied on the surface of concrete with recycled aggregate to create a protective layer. To evaluate the effect of the treatments, tests of water absorption by capillarity and under low pressure with Karsten tubes were performed. Furthermore, SEM-EDS analysis showed the physical barrier caused by biotreatments that produced a reduction on capillarity water absorption of up to 20% and improved the impermeability of recycled concrete against the penetration of water under pressure up to 2.7 times relative to the reference. Therefore, this bioproduct shown to be a promising treatment to protect against penetration of water to concrete surfaces increasing its durability and useful life. Full article

The use of polyhydroxyalkanoates (PHA) as slow-release electron donors for environmental remediation represents a novel and appealing application that is attracting considerable attention in the scientific community. In this context, here, the fermentation pattern of different types of PHA-based materials has been investigated in batch and continuous-flow experiments. Along with commercially available materials, produced from axenic microbial cultures, PHA produced at pilot scale by mixed microbial cultures (MMC) using waste feedstock have been also tested. As a main finding, a rapid onset of volatile fatty acids (VFA) production was observed with a low-purity MMC-deriving material, consisting of microbial cells containing 56% (on weight basis) of intracellular PHA. Indeed, with this material a sustained, long-term production of organic acids (i.e., acetic, propionic, and butyric acids) was observed. In addition, the obtained yield of conversion into acids (up to 70% gVFA/gPHA) was higher than that obtained with the other tested materials, made of extracted and purified PHA. These results clearly suggest the possibility to directly use the PHA-rich cells deriving from the MMC production process, with no need of extraction and purification procedures, as a sustainable and effective carbon source bringing remarkable advantages from an economic and environmental point of view. Full article

Polyhydroxyalkanoates (PHA) are biodegradable polymers that can be intracellularly produced by microorganisms valorizing organic-rich wastes. In the present study, a PHA production system was fed with mussel cooker wastewater after acidogenic fermentation. Besides low pH (4.0 ± 0.3) and high salt (21.7 ± 2.9 g NaCl/L) concentrations, this wastewater also contained nitrogen concentrations (0.8 ± 0.1 g N/L), which were previously reported to be a challenge to the PHA accumulating bacteria enrichment. Bacteria with a PHA storage capacity were selected in an enrichment sequencing batch reactor (SBR) after 60 days of operation. The enriched mixed microbial culture (MMC) was mainly formed by microorganisms from phylum Bacteroidetes, and genera Azoarcus, Comamonas and Thauera from phylum Proteobacteria. The MMC was able to accumulate up to 25 wt% of PHA that was mainly limited by the wastewater nitrogen content, which promoted biomass growth instead of PHA accumulation. Indeed, when the presence of nutrient was limited, PHA stored in the accumulation reactor increased to up to 40.9 wt%. This work demonstrated the feasibility of the enrichment of a MMC with a PHA storage ability valorizing the fish-canning industrial wastewater at low pH, which is generally difficult to treat in wastewater treatment plants. Full article

A “cradle-to-grave” life cycle assessment is performed to identify the environmental issues of polyhydroxyalkanoates (PHAs) produced through a hybrid thermochemical-biological process using anaerobically digested sewage sludge (ADSS) as feedstock. The assessment includes a measure of the energy performance of the process. The system boundary includes: (i) Sludge pyrolysis followed by volatile fatty acids (VFAs) production; (ii) PHAs-enriched biomass production using a mixed microbial culture (MMC); (iii) PHAs extraction with dimethyl carbonate; and iv) PHAs end-of-life. Three scenarios differing in the use of the syngas produced by both pyrolysis and biochar gasification, and two more scenarios differing only in the external energy sources were evaluated. Results show a trade-off between environmental impacts at global scale, such as climate change and resources depletion, and those having an effect at the local/regional scale, such as acidification, eutrophication, and toxicity. Process configurations based only on the sludge-to-PHAs route require an external energy supply, which determines the highest impacts with respect to climate change, resources depletion, and water depletion. On the contrary, process configurations also integrating the sludge-to-energy route for self-sustainment imply more onsite sludge processing and combustion; this results in the highest values of eutrophication, ecotoxicity, and human toxicity. There is not a categorical winner among the investigated configurations; however, the use of a selected mix of external renewable sources while using sludge to produce PHAs only seems the best compromise. The results are comparable to those of both other PHAs production processes found in the literature and various fossil-based and bio-based polymers, in terms of both non-biogenic GHG emissions and energy demand. Further process advancements and technology improvement in high impact stages are required to make this PHAs production process a competitive candidate for the production of biopolymers on a wide scale. Full article

Polyhydroxyalkanoates (PHAs) form a highly promising class of bioplastics for the transition from fossil fuel-based plastics to bio-renewable and biodegradable plastics. Mixed microbial consortia (MMC) are known to be able to produce PHAs from organic waste streams. Knowledge of key-microbes and their characteristics in PHA-producing consortia is necessary for further process optimization and direction towards synthesis of specific types of PHAs. In this study, a PHA-producing mixed microbial consortium (MMC) from an industrial pilot plant was characterized and further enriched on acetate in a laboratory-scale selector with a working volume of 5 L. 16S-rDNA microbiological population analysis of both the industrial pilot plant and the 5 L selector revealed that the most dominant species within the population is Thauera aminoaromatica MZ1T, a Gram-negative beta-proteobacterium belonging to the order of the Rhodocyclales. The relative abundance of this Thauera species increased from 24 to 40% after two months of enrichment in the selector-system, indicating a competitive advantage, possibly due to the storage of a reserve material such as PHA. First experiments with T. aminoaromatica MZ1T showed multiple intracellular granules when grown in pure culture on a growth medium with a C:N ratio of 10:1 and acetate as a carbon source. Nuclear magnetic resonance (NMR) analyses upon extraction of PHA from the pure culture confirmed polyhydroxybutyrate production by T. aminoaromatica MZ1T. Full article

The critical step in the mixed microbial cultures (MMC) polyhydroxyalkanoates (PHA) production process is the selection of an MMC enriched in an efficient PHA-accumulating organism, usually requiring growth-nutrients supplementation. This study aimed at assessing cheese whey (CW) proteins as a source of nitrogen for PHA-producing MMC, thus eliminating or reducing the need for nutrients supplementation. The selection reactor, operated with fermented CW, under a feast–famine regime, was initially supplemented with ammonia–nitrogen in a C/N ratio of 100/15 (Cmol/Nmmol), which was gradually reduced until eliminated, in order to select a culture capable of using CW proteins nitrogen. Decreasing nitrogen supplementation from a C/N ratio of 100/10 to 100/7.5 lead to significant microbial community changes, and reduced the MMC PHA-storing capacity, storage yield, and PHA productivity, decreasing by 30%, and over 45%, respectively. The PHA-storing capacity further deteriorated as nitrogen supplementation was reduced, despite increased protein-uptake. Results show that a culture enriched in PHA-accumulators capable of using proteins as sole nitrogen source could not be attained. In conclusion, this work reports for the first time that an easily bioavailable nitrogen source is required for efficiently selecting PHA-accumulating cultures. Based on the results obtained from this work, a pilot scale plant (two reactors of 100 L) fed with cheese whey for production of PHA is currently being successfully operated under the scope of the YPACK EU project, in which the selection reactor is supplemented with nitrogen (ammonia) from a total C/N ratio of 100/10. Full article