Genome scale metabolic modeling reveals the metabolic potential of three Type II methanotrophs of the genus Methylocystis

Producción CientíficaGenome Scale Metabolic Models (GSMMs) of the recently sequenced Methylocystis hirsuta and two other methanotrophs from the genus Methylocystis have been reconstructed. These organisms are Type II methanotrophs with the ability of accumulating Polyhydroxyalkanoates under nutrient limiting conditions. For the first time, GSMMs have been reconstructed for Type II methanotrophs. These models, combined with experimental biomass and PHB yields of Methylocystis hirsuta, allowed elucidating the methane oxidation mechanism by the enzyme pMMO (particulate methane monooxygenase) in these organisms. In contrast to Type I methanotrophs, which use the “direct coupling mechanism”, Type II methanotrophs appear to use the so called “redox arm mechanism”. The utilization of the “redox arm mechanism”, which involves the coupling between methane oxidation and complex I of the respiratory chain, was confirmed by inhibition of complex I with catechol. Utilization of the “redox arm” mechanism leads to lower biomass yields on methane compared to Type I methanotrophs. However, the ability of Type II methanotrophs to redirect high metabolic carbon fluxes towards acetoacetyl-CoA under nitrogen limiting conditions makes these organisms promising platforms for metabolic engineering.Marie Curie grant H2020-MSCA-IF-2016 CH4BioVal (GA nº 750126).Junta de Castilla y León (Ref. Project VA281P18)Ministerio de Ciencia e Innovación (Proyect CLU 2017-09, CTM2015-70442-R

Continuous polyhydroxybutyrate production from biogas in an innovative two‐stage bioreactor configuration

Producción CientíficaBiogas biorefineries have opened up new horizons beyond heat and electricityproduction in the anaerobic digestion sector. Added‐value products such aspolyhydroxyalkanoates (PHAs), which are environmentally benign and potentialcandidates to replace conventional plastics, can be generated from biogas. Thiswork investigated the potential of an innovative two‐stage growth‐accumulationsystem for the continuous production of biogas‐based polyhydroxybutyrate (PHB)usingMethylocystis hirsutaCSC1 as cell factory. The system comprised twoturbulent bioreactors in series to enhance methane and oxygen mass transfer: acontinuous stirred tank reactor (CSTR) and a bubble column bioreactor (BCB) withinternal gas recirculation. The CSTR was devoted to methanotrophic growth undernitrogen balanced growth conditions and the BCB targeted PHB productionunder nitrogen limiting conditions. Two different operational approaches underdifferent nitrogen loading rates and dilution rates were investigated. A balancednitrogen loading rate along with a dilution rate (D) of 0.3 day−1resulted in the moststable operating conditions and a PHB productivity of ~53 g PHB m−3day−1.However, higher PHB productivities (~127 g PHB m−3day−1)wereachievedusingnitrogen excess at a D = 0.2 day−1. Overall, the high PHB contents (up to 48% w/w)obtained in the CSTR under theoretically nutrient balanced conditions and the poorprocess stability challenged the hypothetical advantages conferred by multistagevs single‐stage process configurations for long‐term PHB production.Ministerio de Ciencia e Innovación under (BES-2016-077160) contract and (project CTM2015-70442-R)Junta de Castilla y León and EU-FEDER program (UIC 315, CLU 2017-09

Toluene biodegradation in an algal-bacterial airlift photobioreactor: Influence of the biomass concentration and of the presence of an organic phase

Producción CientíficaThe potential of algal-bacterial symbiosis for off-gas abatement was investigated for the first time by comparatively evaluating the performance of a bacterial (CB) and an algal-bacterial (PB) airlift bioreactors during the treatment of a 6 g m−3 toluene laden air emission. The influence of biomass concentration and of the addition of a non-aqueous phase was also investigated. A poor and fluctuating performance was recorded during the initial stages of the experiment, which was attributed to the low biomass concentration present in both reactors and to the accumulation of toxic metabolites. In this sense, an increase in the dilution rate from 0.23 to 0.45 d−1 and in biomass concentration from ∼1 to ∼5 g L−1 resulted in elimination capacities (ECs) of 300 g m−3 h−1 (corresponding to removal efficiencies ∼ 90%). Microalgae activity allowed for a reduction in the emitted CO2 and an increase in dissolved O2 concentration in the PB. However, excess biomass growth over 11 g L−1 hindered light penetration and severely decreased photosynthetic activity. The addition of silicone oil at 20% (on a volume basis) stabilized system performance, leading to dissolved O2 concentrations of 7 mg L−1 and steady ECs of 320 g m−3 h−1 in the PB. The ECs here recorded were considerably higher than those previously reported in toluene-degrading bioreactors. Finally, microbial population analysis by DGGE-sequencing demonstrated the differential specialization of the microbial community in both reactors, likely resulting in different toluene degradation pathways and metabolites production.Ministerio de Economía, Industria y Competitividad (Proyect CTM2015-70442-R)Junta de Castilla y León (programa de apoyo a proyectos de investigación – Ref. VA024U14 y UIC 71

Optimization of nitrogen feeding strategies for improving polyhydroxybutyrate production from biogas by Methylocystis parvus OBBP in a stirred tank reactor

Producción CientíficaThe design of efficient cultivation strategies to produce bioplastics from biogas is crucial for the implementation of this biorefinery process. In this work, biogas-based polyhydroxybutyrate (PHB) production and CH4 biodegradation performance was investigated for the first time in a stirred tank bioreactor inoculated with Methylocystis parvus OBBP. Decreasing nitrogen loading rates in continuous mode and alternating feast:famine regimes of 24 h-cycles, and alternating feast:famine regimes of 24 h:24 h and 24 h:48 h were tested. Continuous N feeding did not support an effective PHB production despite the occurrence of nitrogen limiting conditions. Feast-famine cycles of 24 h:24 h (with 50% stoichiometric nitrogen supply) supported the maximum PHB production (20 g-PHB m−3 d−1) without compromising the CH4-elimination capacity (25 g m−3 h−1) of the system. Feast:famine ratios ≤1:2 entailed the deterioration of process performance at stoichiometric nitrogen inputs ≤60%.Ministerio de Ciencia, Innovación y Universidades (grant BES-2016-077160)Ministerio de Ciencia, Innovación y Universidades (project CTM2015-70442-R)Junta de Castilla y León (projects UIC 071 and CLU 2017–09

Ectoine production from biogas in waste treatment facilities: A techno-economic and sensitivity analysis

Producción CientíficaThe capacity of haloalkaliphilic methanotrophic bacteria to synthesize ectoine from CH4-biogas represents an opportunity for waste treatment plants to improve their economic revenues and align their processes to the incoming circular economy directives. A techno-economic and sensitivity analysis for the bioconversion of biogas into 10 t ectoine·y–1 was conducted in two stages: (I) bioconversion of CH4 into ectoine in a bubble column bioreactor and (II) ectoine purification via ion exchange chromatography. The techno-economic analysis showed high investment (4.2 M€) and operational costs (1.4 M€·y–1). However, the high margin between the ectoine market value (600–1000 €·kg–1) and the estimated ectoine production costs (214 €·kg–1) resulted in a high profitability for the process, with a net present value evaluated at 20 years (NPV20) of 33.6 M€. The cost sensitivity analysis conducted revealed a great influence of equipment and consumable costs on the ectoine production costs. In contrast to alternative biogas valorization into heat and electricity or into low added-value bioproducts, biogas bioconversion into ectoine exhibited high robustness toward changes in energy, water, transportation, and labor costs. The worst- and best-case scenarios evaluated showed ectoine break-even prices ranging from 158 to 275 €·kg–1, ∼3–6 times lower than the current industrial ectoine market value.Junta de Castilla y León - Fondo Europeo de Desarrollo Regional (project CLU 2017-09, UIC 315)Junta de Castilla y León - Universidad de Valladolid (contract C18IPJCL)European Union’s Horizon 2020 research and innovation program. grant agreement no. 83799

Polyhydroxyalkanoates production from methane emissions in Sphagnum mosses: Assessing the effect of temperature and phosphorus limitation

Producción CientíficaThe isolation of highly efficient methanotrophic communities is crucial for the optimization of methane bioconversion into products with a high market value such as polyhydroxyalkanoates (PHA). The research here presented aimed at enriching a methanotrophic consortium from two different inocula (Sphagnum peat moss (Sp) and Sphagnum and activated sludge (M)) able to accumulate PHA while efficiently oxidizing CH4. Moreover, the effect of the temperature and phosphorus limitation on the biodegradation rate of CH4 and the PHA accumulation potential was investigated. Higher CH4 degradation rates were obtained under P availability at increasing temperature (25, 30 and 37 °C). The biomass enriched from the mixed inoculum always exhibited a superior biodegradation performance regardless of the temperature (a maximum value of 84.3 ± 8.4 mg CH4 h−1 g biomass−1 was recorded at 37 °C). The results of the PHB production showed that phosphorus limitation is required to promote PHB accumulation, the highest PHB content being observed with the Sphagnum inoculum at 25 °C (13.6 ± 5.6%). The differential specialization of the microbial communities depending on the enrichment temperature supported the key role of this parameter on the results obtained. In all cases after the completion of the enrichment process and of the P limitation tests, Methylocystis, a type II methanotroph known for its ability to accumulate PHA, was the genus that became dominant (reaching percentages from 16 to 46% depending on the enrichment temperature). Thus, the results here obtained demonstrated for the first time the relevance of the temperature used for the enrichment of the methanotrophic bacteria to boost PHA production yields under P limiting condition, highlighting the importance of optimizing culture conditions to improve the cost-efficiency of bioprocesses based on using methane as the primary feedstock for the PHA industrial market.Ministerio de Economía, Industria y Competitividad - FEDER (Project CTM2015-73228-JIN)Junta de Castilla y León (programa de apoyo a proyectos de investigación - Ref. UIC 71 and CLU 2017-09

Assessing the influence of the carbon source on the abatement of industrial N2O emissions coupled with the synthesis of added-value bioproducts

Producción CientíficaThe continuous abatement of a synthetic N2O emission from a nitric acid plant coupled with the simultaneously production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) copolymer and the coenzyme Q10 (CoQ10) in a bubble column reactor (BCR) was tested using methanol, glycerol and a mixture of sodium acetate-acetic acid (Ac-HAc) as a carbon and electron donor source. The BCRs were inoculated with Paracoccus denitrificans and supplied with the carbon/electron donor at a loading rate of 139 g C m− 3 d− 1. High N2O removal efficiencies (81–91%) were achieved, with glycerol supporting the highest abatement. The PHBV cell content ranged from 25 to 53%, with highest accumulation in the culture obtained with methanol and Ac-HAc. However, the greatest PHBV productivities were observed in the BCRs operated with glycerol and Ac-HAc (21.7 and 33.5 g PHBV m− 3 d− 1, respectively). Glycerol supply induced the highest molar ratio (23%) of the homopolymer 3-hydroxyvalerate in the composition of PHBV. In addition, the specific cell content of CoQ10 ranged from 0.4 to 1 mg g− 1. This work constitutes, to the best of our knowledge, the first study combining N2O abatement with the simultaneous production of multiple bioproducts, which pave the way to the development of greenhouse gas biorefineries for climate change mitigation.Ministerio de Economía, Industria y Competitividad (Project CTM2015-70442-R and Red NOVEDAR

Indoor air VOCs biofiltration by bioactive coating packed bed bioreactors

Producción CientíficaBioactive coatings are envisaged as a promising biotechnology to tackle the emerging problem of indoor air pollution. This solution could cope with the low concentrations, the wide range of compounds and the hydrophobicity of some indoor air VOCs, which are the most important bottlenecks regarding the implementation of conventional biotechnologies for indoor air treatment. A bioactive coating-based bioreactor was tested in this study for the abatement of different VOCs (n-hexane, toluene and α-pinene) at different empty bed residence times (EBRT) and inlet VOC concentrations. The performance of this reactor was compared with a conventional biofilm-based bioreactor operated with the same microbial inoculum. After an acclimation period, the bioactive coating-based bioreactor achieved abatements of over 50% for hexane, 80% for toluene and 70% for pinene at EBRTs of 112–56 s and inlet concentrations of 9–15 mg m−3. These results were about 25, 10 and 20% lower than the highest removals recorded in the biofilm-based bioreactor. Both bioreactors experienced a decrease in VOC abatement by ∼25% for hexane, 45% for toluene and 40% for pinene, after reducing the EBRT to 28 s. When inlet VOC concentrations were progressively reduced, VOC abatement efficiencies did not improve. This fact suggested that low EBRTs and low inlet VOCs concentration hindered indoor air pollutant abatement as a result of a limited mass transfer and bioavailability. Metagenomic analyses showed that process operation with toluene, hexane and pinene as the only carbon and energy sources favored an enriched bacterial community represented by the genera Devosia, Mesorhizobium, Sphingobacterium and Mycobacterium, regardless of the bioreactor configuration. Bioactive coatings were used in this work as packing material of a conventional bioreactor, achieving satisfactory VOC abatement similar to a conventional bioreactor.Ministerio de Ciencia e Innovación y Ministerio de Universidades [project RTI2018-0-096441-B-I00]Junta de Castilla y León - EU-FEDER [grant number CLU 2017–09, CL-EI-2021-07 y UIC 315]Junta de Castilla y León y Fondo Social Europeo (PhD grant BDNS 487971)Comisión Europea-H2020-MSCA–IF–2019 (ENHANCEMENT, ref. 897284

Influence of liquid-to-biogas ratio and alkalinity on the biogas upgrading performance in a demo scale algal-bacterial photobioreactor

The influence of the liquid-to-biogas ratio (L/G) and alkalinity on methane quality was evaluated in a 11.7 m3 outdoors horizontal semi-closed tubular photobioreactor interconnected to a 45-L absorption column (AC). CO2 concentrations in the upgraded methane ranged from <0.1 to 9.6% at L/G of 2.0 and 0.5, respectively, with maximum CH4 concentrations of 89.7% at a L/G of 1.0. Moreover, an enhanced CO2 removal (mediating a decrease in CO2 concentration from 9.6 to 1.2%) and therefore higher CH4 contents (increasing from 88.0 to 93.2%) were observed when increasing the alkalinity of the AC cultivation broth from 42 ± 1 mg L−1 to 996 ± 42 mg L−1. H2S was completely removed regardless of the L/G or the alkalinity in AC. The continuous operation of the photobioreactor with optimized operating parameters resulted in contents of CO2 (<0.1%–1.4%), H2S (<0.7 mg m−3) and CH4 (94.1%–98.8%) complying with international regulations for methane injection into natural gas grids.Peer ReviewedPostprint (published version

Treatment of Social Media in Person Name Disambiguation in the Web

En este trabajo presentamos dos heurísticas para tratar páginas web correspondientes a redes sociales en el problema de desambiguación de nombres de persona en la Web. Este problema consiste en agrupar las páginas web proporcionadas por un motor de búsqueda al consultar un nombre de persona según el individuo al que se refieren. Aunque estas páginas web pueden afectar negativamente en la agrupación de los resultados, la mayoría de sistemas del estado del arte no tienen en cuenta su papel en este problema. Hemos evaluado nuestras heurísticas con dos colecciones que contienen este tipo de páginas web. Para agrupar las páginas web hemos utilizado una extensión de un algoritmo del estado del arte. Ambas heurísticas obtienen mejoras cuando hay un número elevado de páginas sociales y el algoritmo propuesto es más independiente del nivel de ambigüedad de los nombres de persona que otros propuestos por el estado del arte.In this work, we present two heuristics to treat web pages from social networks for person name disambiguation in the Web. This problem consists in clustering the results provided by a search engine when the query is a person name according to the individual they refer to. Although these web pages could negatively affect when grouping the results, most of the systems in the state-of-the-art do not take into account their role in this problem. We have evaluated our heuristics with two collections that contain this kind of web pages. We have used an extension of an algorithm of the state of the art to cluster the web pages. Both heuristics get improvements when there is a high number of social web pages, and the proposed algorithm is more independent with respect to the ambiguity degree of person names than other ones in the state of the art.Este trabajo ha sido subvencionado por el Ministerio de Ciencia e Innovación [MED-RECORD Project, TIN2013-46616-C2-2-R] y el grupo CVIP de la URJC