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From CO2 to CH4 via biological methanation
No full textThe leitmotif of this Ph.D. thesis is represented by carbon dioxide (CO2) recycling via biological production of methane (CH4).
This work particularly focuses on the physiology of three hydrogenotrophic methanogens, Methanothermobacter marburgensis (M. marburgensis), Methanothermococcus okinawensis (M. okinawensis) and Methanococcus maripaludis S2 (M. maripaludis), that can be used as catalysts for biological methane production (BMP) process. This CO2 recycling method is challenging due to an inefficient transfer rate of molecular hydrogen (H2) from the gas phase to the liquid phase. Thus, the biocatalyst performance is limited by H2 availability in the liquid medium. However, several factors, as strain type and media requirements, operating conditions, and reactor design, can contribute to the success of CO2 conversion to CH4. Understanding the physiology of methanogens is a powerful tool for developing a scalable BMP process. Therefore, a novel study on the role of trace metals in pure cultures of M. okinawensis and M. marburgensis respectively is herein proposed. Experimental method of this study included an in silico analysis, closed batch, and fed-batch cultivations. In silico analysis revealed genomic differences among the transport systems and enzymes related to the methanogenesis pathway of these two methanogens. The importance of Fe as metal cofactor in methanogenesis emerged from the in silico analysis and it has been confirmed by the closed batch and fed-batch experiments. M. okinawensis responded to rising concentrations of trace element (TE) by increasing specific growth rate (μ, h-1) and volumetric productivity of methane (MER, mmolL-1h-1) during closed batch cultivation. Furthermore, M. okinawensis shown growth and CH4 in fed-batch cultivation. On the base of fed-batch cultures results, M. marburgensis was prioritized and applied for CO2-based BMP process optimization. It has been proposed a new feeding strategy based on exponential fed-batch cultivation where different medium-, TE- and sulphide dilution rates combinations, and different CO2/H2 inflow rates corresponded to a defined run. The specific setting of each run produced different responses from M. marburgensis. In this context, a MER of 476 mmol L-1 h-1 and μ of 0.69 h-1 were eventually achieved at highest H2/CO2 gassing rate and ratio. However, if these factors mitigate the limitation due to the H2 mass transfer on one side, they also reduce CH4 purity in the offgas on the other side. The combined effect of increasing TE dilution and H2/CO2 gassing rates positively affected the biomass and biomass concentration. Among trace elements, there are heavy metals whose toxicity is higher than others. Heavy metals can seriously affect the functionality of microorganisms, and therefore compromise their performances as biocatalysts of a bio-based process. Not only metals, but also organic compounds, such as carboxylic acids, can damage cells survival. Thus, the second experimental part of this thesis deals with inhibition studies on pure culture of M. maripaludis in closed batch cultivation. Despite the potential applications of M. maripaludis, the knowledge surrounding this strain runs out of lab-scale studies concerning the physiology and toxicology of heavy metals and VFAs. Therefore, M. maripaludis growth and productivity were tested by using copper (Cu), zinc (Zn), acetate (Ac) and propionic acid (Pr) as potential inhibitors of microbial activity. The culture was totally inhibited at concentration of 30, 70 and 100 mgL-1 of Cu and 0.7 and 1 gL-1 of Zn. However, M. maripaludis shows tolerance to 3, 7 and 10 mgL-1 of Cu with different extent. The addition of 0.3 gL-1 of Zn to the medium, rather promoted the biomass build-up of M. maripaludis and cancelled the effect of Cu when used together in the medium. In this study, it has been supported that the inhibition by Cu is due to a reduced or suppressed activity of the CODH/ACS complex producing acetyl-CoA intermediate. Acetyl-CoA is the precursor of many metabolic subsystems (e.g. lipid, amino acids, nucleotides pathways) and its alteration would interfere with them. While CODH/ACS activity is supported by CO2 and methanogenesis intermediate, the other way to produce acetyl-CoA is based on the acetate:CoA ligase. The relevance and the tolerance to rising concentrations of Ac and Pr was also investigated and quantified via HPLC analysis. Concentration of 5 and 10 mgL-1 of acetate did not inhibit nor growth neither productivity. Interestingly, the deprivation of acetate not only impacted on the growth rate but also on methanogenesis in M. maripaludis. In absence of Ac, the same concentrations of Pr caused a slow-down of the growth, while productivity was not touched. This study sheds light on the individual and combined impact of Cu, Zn, acetate and propionic acid on the metabolism of M. maripaludis. Furthermore, an attempt to define a possible mechanism which regulates specific acetate capture is provided in this study and the relevance of acetate:CoA ligase respect to CODH/ACS complex for acetyl-CoA synthesis is herein discussed. The information collected in this study are essential to improve the process efficiency of CO2 conversion to CH4 and extend the knowledge on the physiology of certain compounds. The tendency of these methanogens to adapt to adverse conditions, most of the time, offers the possibility to improve the engineering aspects of a limited process toward an unlimited one. Moreover, as a future activity, this thesis proposed the use of a 10-bar pressure bioreactor which has been projected in the frame of the Ph.D. research with a view to improving the success of biological CH4 production
A Microbial-Centric View of Mobile Phones: Enhancing the Technological Feasibility of Biotechnological Recovery of Critical Metals
Get PDFEnd-of-life (EoL) mobile phones represent a valuable reservoir of critical raw materials at higher concentrations compared to primary ores. This review emphasizes the critical need to transition from single-material recovery approaches to comprehensive, holistic strategies for recycling EoL mobile phones. In response to the call for sustainable techniques with reduced energy consumption and pollutant emissions, biohydrometallurgy emerges as a promising solution. The present work intends to review the most relevant studies focusing on the exploitation of microbial consortia in bioleaching and biorecovery processes. All living organisms need macro- and micronutrients for their metabolic functionalities, including some of the elements contained in mobile phones. By exploring the interactions between microbial communities and the diverse elements found in mobile phones, this paper establishes a microbial-centric perspective by connecting each element of each layer to their role in the microbial cell system. A special focus is dedicated to the concepts of ecodesign and modularity as key requirements in electronics to potentially increase selectivity of microbial consortia in the bioleaching process. By bridging microbial science with sustainable design, this review proposes an innovative roadmap to optimize metal recovery, aligning with the principles of the circular economy and advancing scalable biotechnological solutions for electronic waste management
Perspective on the use of methanogens in lithium recovery from brines
Get PDF: Methanogenic archaea stand out as multipurpose biocatalysts for different applications in wide-ranging industrial sectors due to their crucial role in the methane (CH4) cycle and ubiquity in natural environments. The increasing demand for raw materials required by the manufacturing sector (i.e., metals-, concrete-, chemicals-, plastic- and lubricants-based industries) represents a milestone for the global economy and one of the main sources of CO2 emissions. Recovery of critical raw materials (CRMs) from byproducts generated along their supply chain, rather than massive mining operations for mineral extraction and metal smelting, represents a sustainable choice. Demand for lithium (Li), included among CRMs in 2023, grew by 17.1% in the last decades, mostly due to its application in rechargeable lithium-ion batteries. In addition to mineral deposits, the natural resources of Li comprise water, ranging from low Li concentrations (seawater and freshwater) to higher ones (salt lakes and artificial brines). Brines from water desalination can be high in Li content which can be recovered. However, biological brine treatment is not a popular methodology. The methanogenic community has already demonstrated its ability to recover several CRMs which are not essential to their metabolism. Here, we attempt to interconnect the well-established biomethanation process with Li recovery from brines, by analyzing the methanogenic species which may be suitable to grow in brine-like environments and the corresponding mechanism of recovery. Moreover, key factors which should be considered to establish the techno-economic feasibility of this process are here discussed
Novel insights in dimethyl carbonate-based extraction of polyhydroxybutyrate (PHB)
Get PDFBackground: Plastic plays a crucial role in everyday life of human living, nevertheless it represents an undeniable
source of land and water pollution. Polyhydroxybutyrate (PHB) is a bio-based and biodegradable polyester, which can
be naturally produced by microorganisms capable of converting and accumulating carbon as intracellular granules.
Hence, PHB-producing strains stand out as an alternative source to fossil-derived counterparts. However, the extraction
strategy affects the recovery efficiency and the quality of PHB. In this study, PHB was produced by a genetically
modified Escherichia coli strain and successively extracted using dimethyl carbonate (DMC) and ethanol as alternative
solvent and polishing agent to chloroform and hexane. Eventually, a Life Cycle Assessment (LCA) study was performed
for evaluating the environmental and health impact of using DMC.
Results: Extraction yield and purity of PHB obtained via DMC, were quantified, and compared with those obtained
via chloroform-based extraction. PHB yield values from DMC-based extraction were similar to or higher than those
achieved by using chloroform (≥ 67%). To optimize the performance of extraction via DMC, different experimental
conditions were tested, varying the biomass state (dry or wet) and the mixing time, in presence or in absence of a
paper filter. Among 60, 90, 120 min, the mid-value allowed to achieve high extraction yield, both for dry and wet
biomass. Physical and molecular dependence on the biomass state and solvent/antisolvent choice was established.
The comparative LCA analysis promoted the application of DMC/ethanol rather than chloroform/hexane, as the best
choice in terms of health prevention. However, an elevated impact score was achieved by DMC in the environmentallike
categories in contrast with a minor contribution by its counterpart.
Conclusion: The multifaceted exploration of DMC-based PHB extraction herein reported extends the knowledge of
the variables affecting PHB purification process. This work offers novel and valuable insights into PHB extraction process,
including environmental aspects not discussed so far. The findings of our research question the DMC as a green
solvent, though also the choice of the antisolvent can influence the impact on the examined categories.This work has been funded by the Horizon 2020 EU Framework Programme:
CELBICON project, Grant agreement number: 679050. Open access funding
provided by PRIME project funded by the POR FESR 2014/2020 Programme,
Asse I – Azione I.1b.2.2 Regione Piemonte, within the Piattaforma Tecnologica
per la Bioeconomia.Publicad
Unlocking the power of underground (bio)methanation
Get PDFL'abstract è presente nell'allegato / the abstract is in the attachmen
The physiology of trace elements in biological methane production
No full textTrace element (TE) requirements of Methanothermobacter okinawensis and Methanothermobacter marburgensis were examined in silico, and using closed batch and fed-batch cultivation experiments. In silico analysis revealed genomic differences among the transport systems and enzymes related to the archaeal Wood-Ljungdahl pathway of these two methanogens. M. okinawensis responded to rising concentrations of TE by increasing specific growth rate (μ) and volumetric productivity (MER) during closed batch cultivation, and can grow and produce methane (CH4) during fed-batch cultivation. M. marburgensis showed higher μ and MER during fed-batch cultivation and was therefore prioritized for subsequent optimization of CO2-based biological CH4 production. Multiple-parameter cultivation dependency on growth and productivity of M. marburgensis was finally examined using exponential fed-batch cultivation at different medium-, TE- and sulphide dilution rates, and different gas inflow rates. MER of 476 mmol L−1 h−1 and μ of 0.69 h−1 were eventually obtained during exponential fed-batch cultivations employing M. marburgensis
“Innovative high pressure/high temperature, multi-sensing bioreactors system for microbial risk assessment in underground hydrogen storage”
Get PDFThis study addresses the microbial risks associated with Underground Hydrogen Storage (UHS), a critical component in the transition towards renewable energy systems, by employing an innovative multi-reactor system (Bio-xplorer) to simulate UHS conditions in two Italian reservoirs. The microbiological risk assessment (MRA) of Reservoir A and B was evaluated by subjecting them to gas mixtures of 10 % H2 and 90 % CH4, and 99 % H2 and 1 % CO2, respectively. In Reservoir A, the stability of pressure and temperature, the negligible optical density, and lack of microbial metabolites suggested a low risk of microbial activation. Molecular analyses confirmed the absence of sulphate- reducing bacteria (SRB) and limited growth of hydrogenotrophic methanogens (HM). Similarly, in Reservoir B, the absence of SRB and limited occurrence of HM indicated a low microbiological risk. Overall, the present work supports the safe and efficient implementation of UHS, a promising mitigation technique for climate change, using an innovative tool for MRA
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