Biological CO2-Methanation: an Approach to Standardization

Power-to-Methane as one part of Power-to-Gas has been recognized globally as one of the key elements for the transition towards a sustainable energy system. While plants that produce methane catalytically have been in operation for a long time, biological methanation has just reached industrial pilot scale and near-term commercial application. The growing importance of the biological method is reflected by an increasing number of scientific articles describing novel approaches to improve this technology. However, these studies are difficult to compare because they lack a coherent nomenclature. In this article, we present a comprehensive set of parameters allowing the characterization and comparison of various biological methanation processes. To identify relevant parameters needed for a proper description of this technology, we summarized existing literature and defined system boundaries for Power-to-Methane process steps. On this basis, we derive system parameters providing information on the methanation system, its performance, the biology and cost aspects. As a result, three different standards are provided as a blueprint matrix for use in academia and industry applicable to both, biological and catalytic methanation. Hence, this review attempts to set the standards for a comprehensive description of biological and chemical methanation processes

Methanofollis propanolicus sp. nov., a novel archaeal isolate from a Costa Rican oil well that uses propanol for methane production

A novel methanogenic strain, CaP3V-MF-L2AT, was isolated from an exploratory oil well from Cahuita National Park, Costa Rica. The cells were irregular cocci, 0.8–1.8 μm in diameter, stained Gram-negative and were motile. The strain utilized H2/CO2, formate and the primary and secondary alcohols 1-propanol and 2-propanol for methanogenesis, but not acetate, methanol, ethanol, 1-butanol or 2-butanol. Acetate was required as carbon source. The novel isolate grew at 25–40 °C, pH 6.0–7.5 and 0–2.5% (w/v) NaCl. 16S rRNA gene sequence analysis revealed that the strain is affiliated to the genus Methanofollis. It shows 98.8% sequence similarity to its closest relative Methanofollis ethanolicus. The G + C content is 60.1 mol%. Based on the data presented here type strain CaP3V-MF-L2AT (= DSM 113321T = JCM 39176T) represents a novel species, Methanofollis propanolicus sp. nov

A novel interdomain consortium from a Costa Rican oil well composed of Methanobacterium cahuitense sp. nov. and Desulfomicrobium aggregans sp. nov.

A novel interdomain consortium composed of a methanogenic Archaeon and a sulfate-reducing bacterium was isolated from a microbial biofilm in an oil well in Cahuita National Park, Costa Rica. Both organisms can be grown in pure culture or as stable co-culture. The methanogenic cells were non-motile rods producing CH4 exclusively from H2/CO2. Cells of the sulfate-reducing partner were motile rods forming cell aggregates. They utilized hydrogen, lactate, formate, and pyruvate as electron donors. Electron acceptors were sulfate, thiosulfate, and sulfite. 16S rRNA sequencing revealed 99% gene sequence similarity of strain CaP3V-M-L2AT to Methanobacterium subterraneum and 98.5% of strain CaP3V-S-L1AT to Desulfomicrobium baculatum. Both strains grew from 20 to 42 °C, pH 5.0–7.5, and 0–4% NaCl. Based on our data, type strains CaP3V-M-L2AT (= DSM 113354 T = JCM 39174 T) and CaP3V-S-L1AT (= DSM 113299 T = JCM 39179 T) represent novel species which we name Methanobacterium cahuitense sp. nov. and Desulfomicrobium aggregans sp. nov

Cell architecture and flagella of hyperthermophilic Archaea

Earlier studies indicated that flagella might play a crucial role in motility, adhesion, and cell-cell contacts of Archaea. Thus, the ultrastructural and functional characterization of flagella and their anchoring in the cell are crucial for understanding the archaeal cell organization in general. To address this topic, Pyrococcus furiosus was chosen as a suitable model organism. However, in the course of this study, morphological changes of this strain, cultured continuously for several years, were demonstrated. These changes resulted in decreased growth and less adhesion of cells. Comparing this strain to another continuous culture and to the wild type strain, it was shown that culturing in the laboratory caused the adaption of cells to an adhesive phenotype. Nevertheless, cell envelope analyses succeeded in the identification of the glutamate dehydrogenase in membrane fractions of P. furiosus. Immuno-localization studies provided evidence that the enzyme is located on the cell surface and a correlation with adhesion was suggested. Besides the different P. furiosus strains, a newly isolated species, Methanocaldococcus sp. KIN24-T80 was analyzed. Results of characterization studies performed herein confirmed the affiliation of the strain to the genus Methanocaldococcus; the distinctiveness from any previously described species was proven by 16S rRNA gene sequence analysis. Ergo, the isolate was described as novel species named Methanocaldococcus villosus referring to its heavy flagellation and its unique striated surface pattern in SEM preparations. Further following the scope of this study, a submembraneous layer below the cytoplasmic membrane was detected in M. villosus and P. furiosus. For both organisms, it was shown to be related to the anchoring of flagella. In the case of M. villosus, it was proven that the layer is associated with a (putative) chemoreceptor array. Moreover, it was demonstrated for P. furiosus, M. villosus, and five other members of the genus Methanocaldococcus that assembly of flagella is induced as a response to surface associated growth, which helps the cells not only to get in tight contact with the solid support but also with each other. Flagella were identified as structural prerequisite for biofilm formation and other components such as pili or surface proteins were shown to be necessary for a more solid adhesion. To sum up, analyses performed in this thesis extend the current knowledge on the cellular and flagellar ultrastructure of Archaea. Based on the data obtained, M. villosus is proposed as a model organism for studying the flagellation and adhesion of archaeal cells

"Methanocaldococcus villosus" sp. nov., a heavily flagellated archaeon that adheres to surfaces and forms cell–cell contacts

A novel chemolithoautotrophic, hyperthermophilic methanogen was isolated from a submarine hydrothermal system at the Kolbeinsey Ridge, north of Iceland. Based on its 16S rRNA gene sequence, the strain belongs to the order Methanococcales within the genus Methanocaldococcus, with approximately 95 % sequence similarity to Methanocaldococcus jannaschii as its closest relative. Cells of the novel organism stained Gram-negative and appeared as regular to irregular cocci possessing more than 50 polar flagella. These cell appendages mediated not only motility but also adherence to abiotic surfaces and the formation of cell–cell contacts. The new isolate grew at 55–90 °C, with optimum growth at 80 °C. The optimum NaCl concentration for growth was 2.5 % (w/v), and the optimal pH was 6.5. The cells gained their energy exclusively by reduction of CO2 with H2. Selenate, tungstate and yeast extract stimulated growth significantly. The genome size was determined to be in the range 1.8–2.0 kb, and the G+C content of the genomic DNA was 30 mol%. Despite being physiologically nearly identical to the other members of the genus Methanocaldococcus, analysis of whole-cell proteins revealed significant differences. Based on the results from phylogenetic, morphological and protein analyses, we conclude that the novel strain represents a novel species of the genus Methanocaldococcus, for which the name Methanocaldococcus villosus sp. nov. is proposed (type strain KIN24-T80T  = DSM 22612T  = JCM 16315T)

Pyrococcus furiosus flagella: biochemical and transcriptional analyses identify the newly detected flaB0 gene to encode the major flagellin

We have described previously that the flagella of the Euryarchaeon Pyrococcus furiosus are multifunctional cell appendages used for swimming, adhesion to surfaces and formation of cell-cell connections. Here, we characterize these organelles with respect to their biochemistry and transcription. Flagella were purified by shearing from cells followed by CsCl-gradient centrifugation and were found to consist mainly of a ca. 30 kDa glycoprotein. Polymerization studies of denatured flagella resulted in an ATP-independent formation of flagella-like filaments. N-terminal sequencing of the main flagellin revealed an unexpected N-terminus. Therefore, we resequenced the respective region of the genome, thereby discovering that the published genome sequence is not correct. A total of 771 bp are missing in the data base, resulting in the fact that a total of three flagellin genes are present. To keep in line with the earlier nomenclature we call these flaB0, flaB1, and flaB2. Very interestingly, the previously not identified flaB0 codes for the major flagellin. Transcriptional analyses of the newly defined flagellar operon identified various different transcripts depending on the growth phase

Combining a robust thermophilic methanogen and packing material with high liquid hold-up to optimize biological methanation in trickle-bed reactors

The hydrogen gas-to-liquid mass transfer is the limiting factor in biological methanation. In trickle-bed reactors, mass transfer can be increased by high flow velocities in the liquid phase, by adding a packing material with high liquid hold-up or by using methanogenic archaea with a high methane productivity. This study developed a polyphasic approach to address all methods at once. Various methanogenic strains and packings were investigated from a microbial and hydrodynamic perspective. Analyzing the ability to produce high-quality methane and to form biofilms, pure cultures of Methanothermobacter performed better than those of the genus Methanothermococcus. Liquid and static hold-up of a packing material and its capability to facilitate attachment was not attributable to a single property. Consequently, it is recommended to carefully match organism and packing for optimized performance of trickle-bed reactors. The ideal combination for the ORBIT system was identified as Methanothermobacter thermoautotrophicus IM5 and DuraTop (R)

Optimized biological CO2-methanation with a pure culture of thermophilic methanogenic archaea in a trickle-bed reactor

In this study, a fully automated process converting hydrogen and carbon dioxide to methane in a high temperature trickle-bed reactor was developed from lab scale to field test level. The reactor design and system performance was optimized to yield high methane content in the product gas for direct feed-in to the gas grid. The reaction was catalyzed by a pure culture of Methanothermobacter thermoautotrophicus IM5, which formed a biofilm on ceramic packing elements. During 600 h in continuous and semi-continuous operation in countercurrent flow, the 0.05 m3 reactor produced up to 95.3 % of methane at a methane production rate of 0.35 m3CH4 mg3h- 1. Adding nitrogen as carrier gas during startup, foam control and dosing of ammonium and sodium sulfide as nitrogen and sulfur source were important factors for process automation

The Iho670 Fibers of Ignicoccus hospitalis: a New Type of Archaeal Cell Surface Appendage ▿

Ignicoccus hospitalis forms many cell surface appendages, the Iho670 fibers (width, 14 nm; length, up to 20 μm), which constitute up to 5% of cellular protein. They are composed mainly of protein Iho670, possessing no homology to archaeal flagellins or fimbrins. Their existence as structures different from archaeal flagella or fimbriae have gone unnoticed up to now because they are very brittle