Phototrophic Purple Bacteria Research Articles

Elucidating metabolic tuning of mixed purple phototrophic bacteria biofilms in photoheterotrophic conditions through microbial photo-electrosynthesis

Reducing greenhouse gas emissions is critical for humanity nowadays, but it can be beneficial by developing engineered systems that valorize CO2 into commodities, thus mimicking nature’s wisdom. Purple phototrophic bacteria (PPB) naturally accept CO2 into their metabolism as a primary redox sink system in photo-heterotrophy. Dedicated use of this feature for developing sustainable processes (e.g., through negative-emissions photo-bioelectrosynthesis) requires a deep knowledge of the inherent metabolic mechanisms. This work provides evidence of tuning the PPB metabolic mechanisms upon redox stressing through negative polarization (−0.4 and −0.8 V vs. Ag/AgCl) in photo-bioelectrochemical devices. A mixed PPB-culture upregulates its ability to capture CO2 from organics oxidation through the Calvin-Besson-Bassam cycle and anaplerotic pathways, and the redox imbalance is promoted to polyhydroxyalkanoates production. The ecological relationship of PPB with mutualist bacteria stabilizes the system and opens the door for future development of photo-bioelectrochemical devices focused on CO up-cycling.

Propionic acid enhances H2 production in purple phototrophic bacteria: Insights into carbon and reducing equivalent allocation

Biohydrogen is gaining popularity as a clean and cost-effective energy source. Among the various production methods, photo fermentation (PF) with purple phototrophic bacteria (PPB) has shown great opportunity due to its high hydrogen yield. In practice, this yield is influenced by several factors, with the carbon source, particularly simple organic acid, being a key element that has attracted considerable research interest. Short-chain volatile fatty acids (VFAs), such as acetate, propionate, and butyrate, are widely found in waste streams and dark fermentation (DF) effluent. However, most studies on these VFAs focus mainly on performance evaluation, with few exploring the underlying mechanisms, which limits their applicability in real-world scenarios. To uncover the metabolic mechanisms, this study uses metagenomics to clarify the processes of reducing power production and distribution during substrate assimilation. Meanwhile, this study presents the impact of short-chain VFAs on biohydrogen, polyhydroxyalkanoates (PHA) and glycogen production by PPB. The results show that: (1) over long-term cultivation at similar COD consumption rates of 0.06 g COD/d, PPB possessed the highest hydrogen yield when fed with propionate (0.620 L H2·g COD-1) compared with butyrate (0.434) and acetate (0.361); (2) with propionate as the substrate, PPB accumulated less PHA (7 % of dry biomass) but more glycogen content (11 %), compared to butyrate (15 % PHA and 8 % glycogen) and acetate (21 % PHA and 5 % glycogen); (3) metagenomic analysis revealed that propionate resulted in the highest amounts of reducing equivalents, followed by butyrate and acetate; hydrogen production was the most efficient pathway for utilizing the reducing power with propionate, as the CO2 fixation and PHA or glycogen synthesis were ineffective for electron dissipation. This study offers insights into metabolic mechanism that could guide waste stream selection and pretreatment processes to provide favorable VFAs for the PF process, thereby enhancing PPB biohydrogen production performance in practical applications.

Light intensity effects on bioproduct recovery from fuel synthesis wastewater using purple phototrophic bacteria in a hybrid biofilm-suspended growth system

This research looked at how three different light intensities (1600, 4300, and 7200 lx) affect the biomass development, treatment of fuel synthesis wastewater and the recovery of valuable bioproducts between biofilm and suspended growth in a purple-bacteria enriched photobioreactor. Each condition was run in duplicate using an agricultural shade cloth as the biofilm support media in a continuously mixed batch reactor. The results showed that the highest chemical oxygen demand (COD) removal rate (56.8 ± 0.9 %) was found under the highest light intensity (7200 lx), which also led to the most biofilm formation and highest biofilm biomass production (1225 ± 95.7 mg). The maximum carotenoids (Crts) and bacteriochlorophylls (BChls) content occurred in the suspended growth of the 7200 lx reactor. BChls decreased with light intensity in suspended growth, while in biofilm both Crts and BChls were relatively stable between light conditions, likely due to an averaging effect as biofilm thickened at higher light intensity. Light intensity did not affect protein content of the biomass, however, biofilm showed a lower average (41.2 % to 43.7 %) than suspended biomass (45.4 % to 47.7 %). For polyhydroxybutyrate (PHB) the highest cell concentration in biofilm occurred at 1600 lx (11.4 ± 2.4 %), while for suspended growth it occurred at 7200 lx (22.7 ± 0.3 %), though total PHB productivity remained similar between reactors. Shading effects from the externally located biofilm could explain most variations in bioproduct distribution. Overall, these findings suggest that controlling light intensity can effectively influence the treatment of fuel synthesis wastewater and the recovery of valuable bioproducts in a biofilm photobioreactor.

Synergistic phenotypic adaptations of motile purple sulphur bacteria Chromatium okenii during lake-to-laboratory domestication.

Isolating microorganisms from natural environments for cultivation under optimized laboratory settings has markedly improved our understanding of microbial ecology. Artificial growth conditions often diverge from those in natural ecosystems, forcing wild isolates into distinct selective pressures, resulting in diverse eco-physiological adaptations mediated by modification of key phenotypic traits. For motile microorganisms we still lack a biophysical understanding of the relevant traits emerging during domestication and their mechanistic interplay driving short-to-long-term microbial adaptation under laboratory conditions. Using microfluidics, atomic force microscopy, quantitative imaging, and mathematical modeling, we study phenotypic adaptation of Chromatium okenii, a motile phototrophic purple sulfur bacterium from meromictic Lake Cadagno, grown under laboratory conditions over multiple generations. Our results indicate that naturally planktonic C. okenii leverage shifts in cell-surface adhesive interactions, synergistically with changes in cell morphology, mass density, and distribution of intracellular sulfur globules, to suppress their swimming traits, ultimately switching to a sessile lifeform. A computational model of cell mechanics confirms the role of such phenotypic shifts in suppressing the planktonic lifeform. By investigating key phenotypic traits across different physiological stages of lab-grown C. okenii, we uncover a progressive loss of motility during the early stages of domestication, followed by concomitant deflagellation and enhanced surface attachment, ultimately driving the transition of motile sulfur bacteria to a sessile state. Our results establish a mechanistic link between suppression of motility and surface attachment via phenotypic changes, underscoring the emergence of adaptive fitness under laboratory conditions at the expense of traits tailored for natural environments.

The phototrophic purple non-sulfur bacteria Rhodomicrobium spp. are novel chassis for bioplastic production.

Petroleum-based plastics levy significant environmental and economic costs that can be alleviated with sustainably sourced, biodegradable, and bio-based polymers such as polyhydroxyalkanoates (PHAs). However, industrial-scale production of PHAs faces barriers stemming from insufficient product yields and high costs. To address these challenges, we must look beyond the current suite of microbes for PHA production and investigate non-model organisms with versatile metabolisms. In that vein, we assessed PHA production by the photosynthetic purple non-sulfur bacteria (PNSB) Rhodomicrobium vannielii and Rhodomicrobium udaipurense. We show that both species accumulate PHA across photo-heterotrophic, photo-hydrogenotrophic, photo-ferrotrophic, and photo-electrotrophic growth conditions, with either ammonium chloride (NH4Cl) or dinitrogen gas (N2) as nitrogen sources. Our data indicate that nitrogen source plays a significant role in dictating PHA synthesis, with N2 fixation promoting PHA production during photoheterotrophy and photoelectrotrophy but inhibiting production during photohydrogenotrophy and photoferrotrophy. We observed the highest PHA titres (up to 44.08 mg/L, or 43.61% cell dry weight) when cells were grown photoheterotrophically on sodium butyrate with N2, while production was at its lowest during photoelectrotrophy (as low as 0.04 mg/L, or 0.16% cell dry weight). We also find that photohydrogenotrophically grown cells supplemented with NH4Cl exhibit the highest electron yields - up to 58.89% - while photoheterotrophy demonstrated the lowest (0.27%-1.39%). Finally, we highlight superior electron conversion and PHA production compared to a related PNSB, Rhodopseudomonas palustris TIE-1. This study illustrates the value of studying non-model organisms like Rhodomicrobium for sustainable PHA production and indicates future directions for exploring PNSB metabolisms.

Anaerobic hybrid-photobioreactor using phototrophic purple and green bacteria treating piggery wastewater with sulfur recovery and biogas polishing

An anaerobic hybrid-photobioreactor (AnHPBR) was performed for biogas polishing during piggery wastewater treatment. AnHPBR was exposed to high (4.45 × 105 lx) and low (450 lx) light intensities for growing purple and green sulfur phototrophs separately in each chamber of AnHBBR for soluble sulfide removals. Four phases of hydraulic retention times (HRT, h): 24, 48, 96, and 96 (with added sulfate) of piggery wastewater were operated consecutively for 351 days. Metagenomic analysis of the AnHPBR biomass confirmed that purple and green bacteria were 63–82 %, of which the species varied by HRT and sulfate loadings. In sum, 96 h HRT for AnHPBR presented the highest efficacies in removals of COD (92 %), sulfate (73 %), and sulfide (59 %). Its biogas contained high methane (75 %) and low hydrogen sulfide (0.3 %). Additionally, AnHPBR produced outflow biosolids with a stable sulfur content of 1.88 %.

Light harvesting in purple bacteria does not rely on resonance fine-tuning in peripheral antenna complexes

The ring-like peripheral light-harvesting complex 2 (LH2) expressed by many phototrophic purple bacteria is a popular model system in biological light-harvesting research due to its robustness, small size, and known crystal structure. Furthermore, the availability of structural variants with distinct electronic structures and optical properties has made this group of light harvesters an attractive testing ground for studies of structure–function relationships in biological systems. LH2 is one of several pigment-protein complexes for which a link between functionality and effects such as excitonic coherence and vibronic coupling has been proposed. While a direct connection has not yet been demonstrated, many such interactions are highly sensitive to resonance conditions, and a dependence of intra-complex dynamics on detailed electronic structure might be expected. To gauge the sensitivity of energy-level structure and relaxation dynamics to naturally occurring structural changes, we compare the photo-induced dynamics in two structurally distinct LH2 variants. Using polarization-controlled 2D electronic spectroscopy at cryogenic temperatures, we directly access information on dynamic and static disorder in the complexes. The simultaneous optimal spectral and temporal resolution of these experiments further allows us to characterize the ultrafast energy relaxation, including exciton transport within the complexes. Despite the variations in PPC molecular structure manifesting as clear differences in electronic structure and disorder, the energy-transport and—relaxation dynamics remain remarkably similar. This indicates that the light-harvesting functionality of purple bacteria within a single LH2 complex is highly robust to structural perturbations and likely does not rely on finely tuned electronic- or electron-vibrational resonance conditions.

Molecular structure and characterization of the Thermochromatium tepidum light-harvesting 1 photocomplex produced in a foreign host

Purple phototrophic bacteria possess light-harvesting 1 and reaction center (LH1-RC) core complexes that play a key role in converting solar energy to chemical energy. High-resolution structures of LH1-RC and RC complexes have been intensively studied and have yielded critical insight into the architecture and interactions of their proteins, pigments, and cofactors. Nevertheless, a detailed picture of the structure and assembly of LH1-only complexes is lacking due to the intimate association between LH1 and the RC. To study the intrinsic properties and structure of an LH1-only complex, a genetic system was constructed to express the Thermochromatium (Tch.) tepidum LH1 complex heterologously in a modified Rhodospirillum rubrum mutant strain. The heterologously expressed Tch. tepidum LH1 complex was isolated in a pure form free of the RC and exhibited the characteristic absorption properties of Tch. tepidum. Cryo-EM structures of the LH1-only complexes revealed a closed circular ring consisting of either 14 or 15 αβ-subunits, making it the smallest completely closed LH1 complex discovered thus far. Surprisingly, the Tch. tepidum LH1-only complex displayed even higher thermostability than that of the native LH1-RC complex. These results reveal previously unsuspected plasticity of the LH1 complex, provide new insights into the structure and assembly of the LH1-RC complex, and show how molecular genetics can be exploited to study membrane proteins from phototrophic organisms whose genetic manipulation is not yet possible.

Ammonia-dependent reducing power redistribution for purple phototrophic bacteria culture-based biohydrogen production

The global energy crisis has intensified the search for sustainable and clean alternatives, with biohydrogen emerging as a promising solution to address environmental challenges. Leveraging photo fermentation (PF) process, purple phototrophic bacteria (PPB) can harness reducing power derived from organic substrates to facilitate hydrogen production. However, existing studies report much lower H2 yields than theoretical value when using acetate as carbon source and ammonia as nitrogen source, primarily attributed to the widely employed pulse-feeding mode which suffers from ammonia inhibition effect on nitrogenase. To address this issue, a continuous feeding mode was applied to avoid ammonia accumulation in this study. On the other hand, other pathways like carbon fixation and polyhydroxyalkanoate (PHA) formation could compete reducing power with H2 production. However, the reducing power allocation under continuous feeding mode is not yet clear. In this study, the reducing power allocation and hydrogen production performance were evaluated under various ammonia loading, using acetate as carbon source and infrared LED at around 50 W·m−2 as light source. The results show that (a) The absence of ammonia resulted in the best performance for hydrogen production, with 44 % of the reducing power distributed to H2 and the highest H2 volumetric productivity, while the allocation of reducing power to hydrogen production stopped when ammonia loading was above 7.6 mg NH4-N·L−1·d−1; (b) when PPB required to eliminate reducing power under ammonia limited conditions, PHA production was the preferred pathway followed by the hydrogen production pathway, but once PHA accumulation reached saturation, hydrogen generation pathway dominated; (c) under ammonia limited conditions, the TCA cycle was more activated rendering higher NADH (i.e. reducing power) production compared with that under ammonia sufficient conditions which was verified by metagenomics analysis, and all the hydrogen production, PHA accumulation and carbon fixation pathways were highly active to dissipate reducing power. This work provides the insight of reducing power distribution and PPB biohydrogen production variated by ammonia loading under continuous feeding mode.

Valorization of Purple Phototrophic Bacteria Biomass Resulting from Photo Fermentation Aimed at Biohydrogen Production.

This study evaluated the feasibility of contextually producing hydrogen, microbial proteins, and polyhydroxybutyrate (PHB) using a mixed culture of purple phototrophic bacteria biomass under photo fermentative conditions. To this end, three consecutive batch tests were conducted to analyze the biomass growth curve and to explore the potential for optimizing the production process. Experimental findings indicated that inoculating reactors with microorganisms from the exponential growth phase reduced the duration of the process. Furthermore, the most effective approach for simultaneous hydrogen production and the valorization of microbial biomass was found when conducting the process during the exponential growth phase of the biomass. At this stage, achieved after 3 days of fermentation, the productivities of hydrogen, PHB, and microbial proteins were measured at 63.63 L/m3 d, 0.049 kg/m3 d, and 0.045 kg/m3 d, respectively. The biomass composition comprised a total intracellular compound percentage of 56%, with 27% representing PHB and 29% representing proteins. Under these conditions, the estimated daily revenue was maximized, amounting to 0.6 $/m3 d.

Structural insights into the unusual core photocomplex from a triply extremophilic purple bacterium, Halorhodospira halochloris.

Halorhodospira (Hlr.) halochloris is a triply extremophilic phototrophic purple sulfur bacterium, as it is thermophilic, alkaliphilic, and extremely halophilic. The light-harvesting-reaction center (LH1-RC) core complex of this bacterium displays an LH1-Qy transition at 1,016 nm, which is the lowest-energy wavelength absorption among all known phototrophs. Here we report the cryo-EM structure of the LH1-RC at 2.42 Å resolution. The LH1 complex forms a tricyclic ring structure composed of 16 αβγ-polypeptides and one αβ-heterodimer around the RC. From the cryo-EM density map, two previously unrecognized integral membrane proteins, referred to as protein G and protein Q, were identified. Both of these proteins are single transmembrane-spanning helices located between the LH1 ring and the RC L-subunit and are absent from the LH1-RC complexes of all other purple bacteria of which the structures have been determined so far. Besides bacteriochlorophyll b molecules (B1020) located on the periplasmic side of the Hlr. halochloris membrane, there are also two arrays of bacteriochlorophyll b molecules (B800 and B820) located on the cytoplasmic side. Only a single copy of a carotenoid (lycopene) was resolved in the Hlr. halochloris LH1-α3β3 and this was positioned within the complex. The potential quinone channel should be the space between the LH1-α3β3 that accommodates the single lycopene but does not contain a γ-polypeptide, B800 and B820. Our results provide a structural explanation for the unusual Qy red shift and carotenoid absorption in the Hlr. halochloris spectrum and reveal new insights into photosynthetic mechanisms employed by a species that thrives under the harshest conditions of any phototrophic microorganism known.

Establishing an efficient membrane bioreactor for simultaneous pollutant removal and purple bacteria production under salinity stress

Recovering resources from wastewater to alleviate the energy crisis has become the prevailing trend of technological development. Purple phototrophic bacteria (PPB), a group of fast-growing microbes, have been widely noticed for their potential in producing value-added products from waste streams. However, saline contents in these waste streams, such as food processing wastewater pose a big challenge, which not only restrain the pollutant removal efficiency, but also hinder the growth of functional microbes. To overcome this, a photo anaerobic membrane bioreactor cultivating PPB (PPB-MBR) was constructed and its performance upon long-term salinity stress was investigated. PPB-MBR achieved desirable pollutants removal performance with the average COD and NH4+ removal efficiency being 87% (±8%, n = 87) and 89% (±10%, n = 87), respectively during long-term exposure to salinity stress of 1–80 g NaCl L−1. PPB were predominant during the entire operation period of 87 days (60%–80%), obtaining maximum biomass yield of 0.67 g biomass g−1 CODremoved and protein productivity of 0.18 g L−1 d−1 at the salinity level of 20 g NaCl L−1 and 60 g NaCl L−1, respectively. The sum of value-added products in proportion to the biomass reached 58% at maximum at the salinity level of 60 g NaCl L−1 with protein, pigments and trehalose contributing to 44%, 8.7%, and 5%, respectively. Based on economic analysis, the most cost-saving scenario treating food processing wastewater was revealed at salinity level of around 20 g NaCl L−1. However, more optimization tools are needed to boost the production efficiency so that the profit from value-added products can outweigh the additional cost by excess salinity in the future implication.

Polyhydroxyalkanoate production in a biofilm by mixed culture phototrophic bacteria

Anoxygenic purple phototrophic bacteria (PPB) were utilised in an 80 L biofilm flat plate photobioreactor to generate polyhydroxyalkanoate (PHA) over 44 cycles, with acetate as feed. Over the cycles, net PHA yield (growth + accumulation) averaged 21% while accumulation yield averaged 55%. Average PHA content was 35 wt% volatile solids (VS), with the majority (>80%) being harvested from the biofilm at 100 gTotal solids (TS) L−1. The PPB microbial population averaged 45% of total population. Detailed cycle studies indicated that PHA content (and yield) peaked at 0.5–1 d into the accumulation stage (peak of 53 wt% VS), suggesting that cycle time optimisation could improve both yield and selection of PHA accumulators. The resulting polymeric material was comprised of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with 95.5 mol% 3-hydroxybutyrate and 4.5 mol% 3-hydroxyvalerate content, but the molecular weight, at >1090 kDa, is unusually high for extracted, microbially produced PHA and a feature of this work.

Polyhydroxyalkanoates production in purple phototrophic bacteria ponds: A breakthrough in outdoor pilot-scale operation

The versatile capacity of purple phototrophic bacteria (PPB) for producing valuable bioproducts has gathered renewed interest in the field of resource recovery and waste valorisation. However, greater knowledge regarding the viability of applying PPB technologies in outdoor, large-scale systems is required. This study assessed, for the first time, the upscaling of the phototrophic polyhydroxyalkanoate (PHA) production technology in a pilot-scale system operated in outdoor conditions. An integrated system composed of two up-flow anaerobic sludge blanket (UASB) reactors (for fermentation of wastewater with molasses), and two high-rate algal ponds retrofitted into PPB ponds, was operated in a wastewater treatment plant under outdoor conditions. UASB's adaptation to the outdoor temperatures involved testing different operational settings, namely hydraulic retention times (HRT) of 48 and 72 h, and molasses fermentation in one or two UASBs. Results have shown that the fermentation of molasses in both UASBs with an increased HRT of 72 h was able to ensure a suitable operation during colder conditions, achieving 3.83 ± 0.63 g CODFermentative Products/L, compared to the 3.73 ± 0.85 g CODFermentative Products/L achieved during warmer conditions (molasses fermentation in one UASB; HRT 48 h). Furthermore, the PPB ponds were operated under a light-feast/dark-aerated-famine strategy and fed with the fermented wastewater and molasses from the two UASBs. The best PHA production was obtained during the summer of 2018 and spring of 2019, attaining 34.7 % gPHA/gVSS with a productivity of 0.11 gPHA L−1 day−1 and 36 % gPHA/gVSS with a productivity of 0.14 gPHA L−1 day−1, respectively. Overall, this study showcases the first translation of phototrophic PHA production technology from an artificially illuminated laboratory scale system into a naturally illuminated, outdoor, pilot-scale system. It also addresses relevant process integration aspects with UASBs for pre-fermenting wastewater with molasses, providing a novel operational strategy to achieve photosynthetic PHA production in outdoor full-scale systems.

Potential of enriched phototrophic purple bacteria for H2 bioconversion into single cell protein

Single cell protein (SCP) has emerged as an alternative protein source, potentially based on the recovery of carbon and nutrients from waste-derived resources as part of the circular economy. From those resources, gaseous substrates have the advantage of an easy sterilization, allowing the production of pathogen-free SCP. Sterile gaseous substrates allow producing pathogen-free SCP. This study evaluated the use of an enriched phototrophic purple bacteria (PPB) consortium for SCP production using H2 and CO2 as electron and C sources. The influence of pH (6.0–8.5), temperature (15–50 °C) and light intensity (0–50 W·m−2) on the growth kinetics and biomass yields was investigated using batch tests. Optimal conditions were found at pH 7, 25 °C and light intensities over 30 W·m−2. High biomass and protein yields were achieved (~ 1 g CODbiomass·g CODH2consumed−1 and 3.9–4.4 g protein·g H2−1) regardless of the environmental conditions, being amongst the highest values reported from gaseous streams. These high yields were obtained thanks to the use of light as a sole energy source by the PPB consortium, allowing a total utilization of H2 for growth. Hydrogen uptake rates varied considerably, with values up to 61 ± 5 mg COD·d−1 for the overall H2 consumption rates and 2.00 ± 0.14 g COD·g COD−1·d−1 for the maximum specific uptake rates under optimal growth conditions. The latter value was estimated using a mechanistic model able to represent PPB growth on H2. The biomass exhibited high protein contents (>50 % w/w) and adequate amino acid profiles, showing its suitability as SCP for feed. PPB were the dominant bacteria during the experiments (relative abundance over 80 % in most tests), with a stable population dominated by Rhodobacter sp. and Rhodopseudomonas sp. This study demonstrates the potential of enriched PPB cultures for H2 bioconversion into SCP.

Environmental life cycle assessment of polyhydroxyalkanoates production by purple phototrophic bacteria mixed cultures

Bioplastics offer a promising sustainable alternative to petroleum-based plastics due to their biodegradability as well as favourable thermal and mechanical properties. Among different types of biobased polymers, the production of polyhydroxyalkanoates (PHA) using purple phototrophic bacteria (PPB) and low-value substrates has gained increasing interest. Despite the momentum, challenges regarding the scalability and environmental feasibility of this biopolymer production pathway remain. In response, this study employs an exploratory LCA approach to quantitatively assesses the potential environmental implications of PHA production in powder form and the joint management of the organic fraction of municipal solid waste (OFMSW) through a novel photobiorefinery system that uses PPB mixed cultures. Environmental impacts were tested under multiple improvement scenarios and benchmarked against the production of conventional fossil-based granulate or unprocessed plastics, including low density polyethylene (LDPE), polyethylene terephthalate (PET) and polyurethane (PU). The photobiorefinery stage was found to have the greatest contribution to the impact categories, particularly due to direct emissions, consumption of electricity and production of extractive chemical agents used. These factors accounted for over 70% of the photobiorefinery impact in all cases. Avoided impacts provided net favourable outcomes in terms of carbon footprint and fossil resources when comparing PHA production to conventional plastics, especially PET and PU, with impact reductions ranging from 30% to 60%, respectively. However, when considering other impact categories like eutrophication, this situation was less favourable. The exploration of alternative scenarios offered significant impact reductions, especially when renewable electricity or an environmentally friendly extraction agent is used. Moreover, minimizing methane losses or co-producing hydrogen in the photobiorefinery had a notably positive effect on the carbon footprint, reducing the impact by more than 2 t of CO2 eq per t of PHA powder compared to the base case. Therefore, the implementation of feasible improvement measures in the short term can position PHA produced by mixed cultures as a sustainable alternative to petroleum-based plastics.

A novel mechanistic modelling approach for microbial selection dynamics: Towards improved design and control of raceway reactors for purple bacteria

Purple phototrophic bacteria (PPB) show an underexplored potential for resource recovery from wastewater. Raceway reactors offer a more affordable full-scale solution on wastewater and enable useful additional aerobic processes. Current mathematical models of PPB systems provide useful mechanistic insights, but do not represent the full metabolic versatility of PPB and thus require further advancement to simulate the process for technology development and control. In this study, a new modelling approach for PPB that integrates the photoheterotrophic, and both anaerobic and aerobic chemoheterotrophic metabolic pathways through an empirical parallel metabolic growth constant was proposed. It aimed the modelling of microbial selection dynamics in competition with aerobic and anaerobic microbial community under different operational scenarios. A sensitivity analysis was carried out to identify the most influential parameters within the model and calibrate them based on experimental data. Process perturbation scenarios were simulated, which showed a good performance of the model.

Achieving Discharge Limits in Single-Stage Domestic Wastewater Treatment by Combining Urban Waste Sources and Phototrophic Mixed Cultures.

This work shows the potential of a new way of co-treatment of domestic wastewater (DWW) and a liquid stream coming from the thermal hydrolysis of the organic fraction of municipal solid waste (OFMSW) mediated by a mixed culture of purple phototrophic bacteria (PPB) capable of assimilating carbon and nutrients from the medium. The biological system is an open single-step process operated under microaerophilic conditions at an oxidative reduction potential (ORP) < 0 mV with a photoperiod of 12/24 h and fed during the light stage only so the results can be extrapolated to outdoor open pond operations by monitoring the ORP. The effluent mostly complies with the discharge values of the Spanish legislation in COD and p-values (<125 mg/L; <2 mg/L), respectively, and punctually on values in N (<15 mg/L). Applying an HRT of 3 d and a ratio of 100:7 (COD:N), the presence of PPB in the mixed culture surpassed 50% of 16S rRNA gene copies, removing 78% of COD, 53% of N, and 66% of P. Furthermore, by increasing the HRT to 5 d, removal efficiencies of 83% of COD, 65% of N, and 91% of P were achieved. In addition, the reactors were further operated in a membrane bioreactor, thus separating the HRT from the SRT to increase the specific loading rate. Very satisfactory removal efficiencies were achieved by applying an HRT and SRT of 2.3 and 3 d, respectively: 84% of COD, 49% of N, and 93% of P despite the low presence of PPB due to more oxidative conditions, which step-by-step re-colonized the mixed culture until reaching >20% of 16S rRNA gene copies after 49 d of operation. These results open the door to scaling up the process in open photobioreactors capable of treating urban wastewater and municipal solid waste in a single stage and under microaerophilic conditions by controlling the ORP of the system.

Performance of fish sludge solubilization and phototrophic bioconversion by purple phototrophic bacteria for nutrient recovery in aquaponic system

Nutrient recovery from fish sludge in aquaponics is crucial to improve the economic output of a system sustainably and hygienically. Currently, fish sludge is treated using conventional anaerobic and aerobic mineralization, which does not allow the recovery of valuable nutrients in fish wastes. In this study, a two-stage approach (named as solubilization process and phototrophic bioconversion) is proposed to convert fish sludge into mineral nutrients and biomass nutrients using purple phototrophic bacteria (PPB), thereby promoting the growth of plants and fish simultaneously in aquaponics. Anaerobic and aerobic solubilization methods are tested to pretreat the fish sludge, generating substrates for PPB. Anaerobic solubilization yields 2.1 times more soluble chemical oxygen demand (SCOD) and 3.7 times more total volatile fatty acid (t-VFA) from fish sludge compared with aerobic solubilization. The anaerobic solubilization effluent indicates a CODt-VFA/SCOD of 60% and a VFA comprising 13.3% acetate and 49.0% propionate for PPB. The phototrophic bioconversion using anaerobic solubilization effluent under the light-anaerobic condition results in the highest biomass yield (0.94 g CODbiomass/g CODremoved) and the highest PPB dominance (Ectothiorhodospira, 58.7%). The anaerobic solubilization and light-anaerobic phototrophic bioconversion achieves 54.1% of carbon recovery efficiency (CRE) (in terms of COD), as well as 44.8% and 91.3% of nutrient recovery efficiency (NRE) for N and P. A novel multiloop aquaponic system combined with PPB-based nutrient recovery is proposed for the reuse of mineral nutrients and PPB biomass generated from fish sludge.

Ammonia impact on the selection of a phototrophic - chemotrophic consortium for polyhydroxyalkanoates production under light-feast / dark-aerated-famine conditions

Phototrophic polyhydroxyalkanoate (PHA) production is an emerging technology for recovering carbon and nutrients from diverse wastewater streams. However, reliable selection methods for the enrichment of PHA accumulating purple phototrophic bacteria (PPB) in phototrophic mixed cultures (PMC) are needed. This research evaluates the impact of ammonia on the selection of a PHA accumulating phototrophic-chemotrophic consortium, towards the enrichment of PHA accumulating PPB. The culture was operated under light-feast/dark-aerated-famine and winter simulated-outdoor conditions (13.2 ± 0.9 °C, transient light, 143.5 W/m2), using real fermented domestic wastewater with molasses as feedstock. Three ammonia supply strategies were assessed: 1) ammonia available only in the light phase, 2) ammonia always present and 3) ammonia available only during the dark-aerated-famine phase. Results showed that the PMC selected under 1) ammonia only in the light and 3) dark-famine ammonia conditions, presented the lowest PHA accumulation capacity during the light period (11.1 % g PHA/g VSS and 10.4 % g PHA/g VSS, respectively). In case 1), the absence of ammonia during the dark-aerated-famine phase did not promote the selection of PHA storing PPB, whereas in case 3) the absence of ammonia during the light period favoured cyanobacteria growth as well as purple sulphur bacteria with increased non-PHA inclusions, resulting in an overall decrease of phototrophic PHA accumulation capacity. The best PHA accumulation performance was obtained with selection under permanent presence of ammonia (case 2), which attained a PHA content of 21.6 % g PHA/g VSS (10.2 Cmmol PHA/L), at a production rate of 0.57 g PHA/L·day, during the light period in the selection reactor. Results in case 2 also showed that feedstock composition impacts the PMC performance, with feedstocks richer in more reduced volatile fatty acids (butyric and valeric acids) decreasing phototrophic performance and leading to acids entering the dark-aerated phase. Nevertheless, the presence of organic carbon in the aerated phase was not detrimental to the system. In fact, it led to the establishment of a phototrophic-chemotrophic consortium that could photosynthetically accumulate a PHA content of 13.2 % g PHA/g VSS (6.7 Cmmol PHA/L) at a production rate of 0.20 g PHA/L·day in the light phase, and was able to further increase that storage up to 18.5 % g PHA/g VSS (11.0 Cmmol PHA/L) at a production rate of 1.35 g PHA/L·day in the dark-aerated period. Furthermore, the light-feast/dark-aerated-famine operation was able to maintain the performance of the selection reactor under winter conditions, unlike non-aerated PMC systems operated under summer conditions, suggesting that night-time aeration coupled with the constant presence of ammonia can contribute to overcoming the seasonal constraints of outdoor operation of PMCs for PHA production.