Search Results (24)

This study investigated the production of poly(3-hydroxybutyrate) (PHB) using mixed microbial cultures (MMCs) with lignocellulosic hydrolysates as a carbon source. Single-factor experiments were conducted to examine the effects of substrate concentration, C/N ratio, and pH on PHB synthesis. The highest PHB yield (612.35 mg/L) was achieved at a substrate concentration of 1700 mg/L (R1700), with an effective C/N ratio of approximately 31 and pH 7.0. Nitrogen limitation and neutral pH were favorable for PHB production. Microbial community analysis via 16S rDNA sequencing revealed Chryseobacterium as the dominant genus in all reactors. These findings provide insights into the efficient conversion of waste lignocellulose into biodegradable PHB using MMCs. Full article

The utility of brewery spent grain (BSG), a byproduct of the beer production process, for the synthesis of polyhydroxyalkanoates (PHAs), is a significant advancement towards sustainable and cost-effective biopolymer production. This paper reviews the upcycling potential of BSG as a substrate for PHA production, utilizing various biotechnological approaches to convert this abundant waste material into high-value biodegradable polymers. Through a comprehensive review of recent studies, we highlight the biochemical composition of BSG and its suitability for microbial fermentation processes. This research delves into different methodologies for PHA production from BSG, including the use of mixed microbial cultures (MMCs) for the synthesis of volatile fatty acids (VFAs), a critical precursor in PHA production, and solid-state fermentation (SSF) techniques. We also examine the optimization of process parameters such as pH, temperature, and microbial concentration through the application of the Doehlert design, revealing the intricate relationships between these factors and their impact on VFA profiles and PHA yields. Additionally, this paper discusses challenges and future perspectives for enhancing the efficiency and economic viability of PHA production from BSG. By harnessing the untapped potential of BSG, this research contributes to the development of a circular economy model, emphasizing waste valorization and the creation of sustainable alternatives to conventional plastics. Full article

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

The biologically-derived polymers polyhydroxyalkanoates (PHAs) are biodegradable and can be considered a valuable alternative to conventional fossil-based plastics. However, upstream and downstream processes for PHA production are characterized by high energy and chemical consumption and are not economically competitive with petroleum-based polymers. Aiming to improve both the environmental and economical sustainability of PHAs production, in this work, corn straw was used as raw material to obtain a mixture of fermentable sugars after microwave-assisted flash hydrolysis (2 min, 0.01 g/L, 50.7% yield). A mixed microbial culture enriched from dairy industry waste was used for fermentation in a shake flask, allowing us to achieve good poly(hydroxy-butyrate-co-hydroxy-valerate) yields (41.4%, after 72 h of fermentation). A scale-up in a stirred tank bioreactor (3 L) gave higher yields (76.3%, after 96 h), allowing in both cases to achieve a concentration of 0.42 g/L in the fermentation medium. The possibility of producing PHAs from agricultural waste using a mixed microbial culture from the food industry with enabling technologies could make the production of biopolymers more competitive. Full article

The use of mixed microbial cultures (MMC) and organic wastes and wastewaters as feed sources is considered an appealing approach to reduce the current polyhydroxyalkanoates (PHAs) production costs. However, this method entails an additional hurdle to the PHAs downstream processing (recovery and purification). In the current work, the effect of a sodium hypochlorite (NaClO) pre-treatment coupled with dimethyl carbonate (DMC) or chloroform (CF) as extraction solvents on the PHAs recovery efficiency (RE) from MMC was evaluated. MMC were harvested from a sequencing batch reactor (SBR) fed with a synthetic prefermented olive mill wastewaster. Two different carbon-sources (acetic acid and acetic/propionic acids) were employed during the batch accumulation of polyhydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from MMC. Obtained PHAs were characterized by ¹H and ¹³C nuclear magnetic resonance, gel-permeation chromatography, differential scanning calorimetry, and thermal gravimetric analysis. The results showed that when a NaClO pre-treatment is not added, the use of DMC allows to obtain higher RE of both biopolymers (PHB and PHBV), in comparison with CF. In contrast, the use of CF as extraction solvent required a pre-treatment step to improve the PHB and PHBV recovery. In all cases, RE values were higher for PHBV than for PHB. 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

Polyhydroxyalkanoate (PHA) recovery from microbial cells relies on either solvent extraction (usually using halogenated solvents) and/or digestion of the non-PHA cell mass (NPCM) by the action of chemicals (e.g., hypochlorite) that raise environmental and health hazards. A greener alternative for PHA recovery, subcritical water (SBW), was evaluated as a method for the dissolution of the NPCM of a mixed microbial culture (MMC) biomass. A temperature of 150 °C was found as a compromise to reach NPCM solubilization while mostly preventing the degradation of the biopolymer during the procedure. Such conditions yielded a polymer with a purity of 77%. PHA purity was further improved by combining the SBW treatment with hypochlorite digestion, in which a significantly lower hypochlorite concentration (0.1%, v/v) was sufficient to achieve an overall polymer purity of 80%. During the procedure, the biopolymer suffered some depolymerization, as evidenced by the lower molecular weight (M_w) and higher polydispersity of the extracted samples. Although such changes in the biopolymer’s molecular mass distribution impact its mechanical properties, impairing its utilization in most conventional plastic uses, the obtained PHA can find use in several applications, for example as additives or for the preparation of graft or block co-polymers, in which low-M_w oligomers are sought. 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

The biosynthesis of polyhydroxyalkanoates (PHAs) from industrial wastes by mixed microbial cultures (MMCs) enriched in PHA-accumulating bacteria is a promising technology to replace petroleum-based plastics. However, the populations’ dynamics in the PHA-accumulating MMCs are not well known. Therefore, the main objective of this study was to address the shifts in the size and structure of the bacterial communities in two lab-scale sequencing batch reactors (SBRs) fed with fish-canning effluents and operated under non-saline (SBR-N, 0.5 g NaCl/L) or saline (SBR-S, 10 g NaCl/L) conditions, by using a combination of quantitative PCR and Illumina sequencing of bacterial 16S rRNA genes. A double growth limitation (DGL) strategy, in which nitrogen availability was limited and uncoupled to carbon addition, strongly modulated the relative abundances of the PHA-accumulating bacteria, leading to an increase in the accumulation of PHAs, independently of the saline conditions (average 9.04 wt% and 11.69 wt%, maximum yields 22.03 wt% and 26.33% SBR-N and SBR-S, respectively). On the other hand, no correlations were found among the PHAs accumulation yields and the absolute abundances of total Bacteria, which decreased through time in the SBR-N and did not present statistical differences in the SBR-S. Acinetobacter, Calothrix, Dyella, Flavobacterium, Novosphingobium, Qipengyuania, and Tsukamurella were key PHA-accumulating genera in both SBRs under the DGL strategy, which was revealed as a successful tool to obtain a PHA-enriched MMC using fish-canning effluents. Full article

Volatile fatty acid (VFA) rich streams from fermentation of organic residuals and wastewater are suitable feedstocks for mixed microbial culture (MMC) Polyhydroxyalkanoate (PHA) production. However, many such streams have low total VFA concentration (1–10 gCOD/L). PHA accumulation requires a flow-through bioprocess if the VFAs are not concentrated. A flow through bioprocess must balance goals of productivity (highest possible influent flow rates) with goals of substrate utilization efficiency (lowest possible effluent VFA concentration). Towards these goals, dynamics of upshift and downshift respiration kinetics for laboratory and pilot scale MMCs were evaluated. Monod kinetics described a hysteresis between the upshift and downshift responses. Substrate concentrations necessary to stimulate a given substrate uptake rate were significantly higher than the concentrations necessary to sustain the attained substrate uptake rate. A benefit of this hysteresis was explored in Monte Carlo based PHA accumulation bioprocess numerical simulations. Simulations illustrated for a potential to establish continuous flow-through PHA production bioprocesses even at a low (1 gCOD/L) influent total VFA concentration. Process biomass recirculation into an engineered higher substrate concentration mixing zone, due to the constant influent substrate flow, enabled to drive the process to maximal possible PHA production rates without sacrificing substrate utilization efficiency. 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 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