CO2 Fixation Pathways Research Articles

The reductive glycine pathway allows autotrophic growth of Desulfovibrio desulfuricans

Six CO2 fixation pathways are known to operate in photoautotrophic and chemoautotrophic microorganisms. Here, we describe chemolithoautotrophic growth of the sulphate-reducing bacterium Desulfovibrio desulfuricans (strain G11) with hydrogen and sulphate as energy substrates. Genomic, transcriptomic, proteomic and metabolomic analyses reveal that D. desulfuricans assimilates CO2 via the reductive glycine pathway, a seventh CO2 fixation pathway. In this pathway, CO2 is first reduced to formate, which is reduced and condensed with a second CO2 to generate glycine. Glycine is further reduced in D. desulfuricans by glycine reductase to acetyl-P, and then to acetyl-CoA, which is condensed with another CO2 to form pyruvate. Ammonia is involved in the operation of the pathway, which is reflected in the dependence of the autotrophic growth rate on the ammonia concentration. Our study demonstrates microbial autotrophic growth fully supported by this highly ATP-efficient CO2 fixation pathway.

Expression of Formate-Tetrahydrofolate Ligase Did Not Improve Growth but Interferes With Nitrogen and Carbon Metabolism of Synechocystis sp. PCC 6803.

The introduction of alternative CO2-fixing pathways in photoautotrophic organism may improve the efficiency of biological carbon fixation such as minimizing the carbon loss due to photorespiration. Here, we analyzed the effects of creating a formate entry point into the primary metabolism of the cyanobacterium Synechocystis sp. PCC 6803. The formate-tetrahydrofolate ligase (FTL) from Methylobacterium extorquens AM1 was expressed in Synechocystis to enable formate assimilation and reducing the loss of fixed carbon in the photorespiratory pathway. Transgenic strains accumulated serine and 3-phosphoglycerate, and consumed more 2-phosphoglycolate and glycine, which seemed to reflect an efficient utilization of formate. However, labeling experiments showed that the serine accumulation was not due to the expected incorporation of formate. Subsequent DNA-microarray analysis revealed profound changes in transcript abundance due to ftl expression. Transcriptome changes were observed in relation to serine and glycine metabolism, C1-metabolism and particularly nitrogen assimilation. The data implied that ftl expression interfered with the signaling the carbon/nitrogen ratio in Synechocystis. Our results indicate that the expression of new enzymes could have a severe impact on the cellular regulatory network, which potentially hinders the establishment of newly designed pathways.

Functional cooperation of the glycine synthase-reductase and Wood-Ljungdahl pathways for autotrophic growth of Clostridium drakei.

Among CO2-fixing metabolic pathways in nature, the linear Wood-Ljungdahl pathway (WLP) in phylogenetically diverse acetate-forming acetogens comprises the most energetically efficient pathway, requires the least number of reactions, and converts CO2 to formate and then into acetyl-CoA. Despite two genes encoding glycine synthase being well-conserved in WLP gene clusters, the functional role of glycine synthase under autotrophic growth conditions has remained uncertain. Here, using the reconstructed genome-scale metabolic model iSL771 based on the completed genome sequence, transcriptomics, 13C isotope-based metabolite-tracing experiments, biochemical assays, and heterologous expression of the pathway in another acetogen, we discovered that the WLP and the glycine synthase pathway are functionally interconnected to fix CO2, subsequently converting CO2 into acetyl-CoA, acetyl-phosphate, and serine. Moreover, the functional cooperation of the pathways enhances CO2 consumption and cellular growth rates via bypassing reducing power required reactions for cellular metabolism during autotrophic growth of acetogens.

Third-generation biorefineries as the means to produce fuels and chemicals from CO2

Concerns regarding petroleum depletion and global climate change caused by greenhouse gas emissions have spurred interest in renewable alternatives to fossil fuels. Third-generation (3G) biorefineries aim to utilize microbial cell factories to convert renewable energies and atmospheric CO2 into fuels and chemicals, and hence represent a route for assessing fuels and chemicals in a carbon-neutral manner. However, to establish processes competitive with the petroleum industry, it is important to clarify/evaluate/identify the most promising CO2 fixation pathways, the most appropriate CO2 utilization models and the necessary productivity levels. Here, we discuss the latest advances in 3G biorefineries. Following an overview of applications of CO2 feedstocks, mainly from flue gas and waste gasification, we review prominent opportunities and barriers in CO2 fixation and energy capture. We then summarize reported CO2-based products and industries, and describe trends and key challenges for future advancement of 3G biorefineries. A shift from sugar-based feedstocks and biomass to the use of atmospheric CO2 for the bioproduction of fuels and chemicals is desirable. This Review describes how microorganisms can be engineered for CO2 fixation and industrial valorization of this key molecule.

Metabolic rewiring of synthetic pyruvate dehydrogenase bypasses for acetone production in cyanobacteria.

SummaryDesigning synthetic pathways for efficient CO2 fixation and conversion is essential for sustainable chemical production. Here we have designed a synthetic acetate‐acetyl‐CoA/malonyl‐CoA (AAM) bypass to overcome an enzymatic activity of pyruvate dehydrogenase complex. This synthetic pathway utilizes acetate assimilation and carbon rearrangements using a methyl malonyl‐CoA carboxyltransferase. We demonstrated direct conversion of CO2 into acetyl‐CoA‐derived acetone as an example in photosynthetic Synechococcus elongatus PCC 7942 by increasing the acetyl‐CoA pools. The engineered cyanobacterial strain with the AAM‐bypass produced 0.41 g/L of acetone at 0.71 m/day of molar productivity. This work clearly shows that the synthetic pyruvate dehydrogenase bypass (AAM‐bypass) is a key factor for the high‐level production of an acetyl‐CoA‐derived chemical in photosynthetic organisms.

Carbon nanomaterials affect carbon cycle-related functions of the soil microbial community and the coupling of nutrient cycles

Many studies have examined changes in soil microbial community structure and composition by carbon nanomaterials (CNMs). Few, however, have investigated their impact on microbial community functions. This study explored how fullerene (C60) and multi-walled carbon nanotubes (M50) altered functionality of an agricultural soil microbial community (Archaea, Bacteria and Eukarya), using microcosm experiments combined with GeoChip microarray. M50 had a stronger effect than C60 on alpha diversity of microbial functional genes; both CNMs increased beta diversity, resulting in functional profiles distinct from the control. M50 exerted a broader, severer impact on microbially mediated nutrient cycles. Together, these two CNMs affected CO2 fixation pathways, microbial degradation of diverse carbohydrates, secondary plant metabolites, lipids and phospholipids, proteins, as well as methanogenesis and methane oxidation. They also suppressed nitrogen fixation, nitrification, dissimilatory nitrogen reduction, eukaryotic assimilatory nitrogen reduction, and anaerobic ammonium oxidation (anammox). Phosphorus and sulfur cycles were less vulnerable; only phytic acid hydrolysis and sulfite reduction were inhibited by M50 but not C60. Network analysis suggested decoupling of nutrient cycles by CNMs, manifesting closer and more hierarchical gene networks. This work reinforces profound impact of CNMs on soil microbial community functions and ecosystem services, laying a path for future investigation in this direction.

Characterizing the microbiomes of Antarctic sponges: a functional metagenomic approach

Relatively little is known about the role of sponge microbiomes in the Antarctic marine environment, where sponges may dominate the benthic landscape. Specifically, we understand little about how taxonomic and functional diversity contributes to the symbiotic lifestyle and aids in nutrient cycling. Here we use functional metagenomics to investigate the community composition and metabolic potential of microbiomes from two abundant Antarctic sponges, Leucetta antarctica and Myxilla sp. Genomic and taxonomic analyses show that both sponges harbor a distinct microbial community with high fungal abundance, which differs from the surrounding seawater. Functional analyses reveal both sponge-associated microbial communities are enriched in functions related to the symbiotic lifestyle (e.g., CRISPR system, Eukaryotic-like proteins, and transposases), and in functions important for nutrient cycling. Both sponge microbiomes possessed genes necessary to perform processes important to nitrogen cycling (i.e., ammonia oxidation, nitrite oxidation, and denitrification), and carbon fixation. The latter indicates that Antarctic sponge microorganisms prefer light-independent pathways for CO2 fixation mediated by chemoautotrophic microorganisms. Together, these results show how the unique metabolic potential of two Antarctic sponge microbiomes help these sponge holobionts survive in these inhospitable environments, and contribute to major nutrient cycles of these ecosystems.

The industrial yeast Pichia pastoris is converted from a heterotroph into an autotroph capable of growth on CO2.

The methylotrophic yeast Pichia pastoris is widely used in the manufacture of industrial enzymes and pharmaceuticals. Like most biotechnological production hosts, P. pastoris is heterotrophic and grows on organic feedstocks that have competing uses in the production of food and animal feed. In a step toward more sustainable industrial processes, we describe the conversion of P. pastoris into an autotroph that grows on CO2. By addition of eight heterologous genes and deletion of three native genes, we engineer the peroxisomal methanol-assimilation pathway of P. pastoris into a CO2 fixation pathway resembling the Calvin-Benson-Bassham cycle, the predominant natural CO2 fixation pathway. The resulting strain can grow continuously with CO2 as a sole carbon source at a µmax of 0.008 h-1. The specific growth rate was further improved to 0.018 h-1 by adaptive laboratory evolution. This engineered P. pastoris strain may promote sustainability by sequestering the greenhouse gas CO2 and by avoiding consumption of an organic feedstock with alternative uses in food production.

Growth of the acetogenic bacterium Acetobacterium woodii by dismutation of acetaldehyde to acetate and ethanol.

Acetogenic bacteria are a group of strictly anaerobic bacteria that may have been first life forms on Earth since they employ an ancient pathway for CO2 fixation into acetyl-CoA that is coupled to the synthesis of ATP, the Wood-Ljungdahl pathway. Electrons for CO2 reduction are derived from oxidation of H2 or CO and thus, these bacteria can grow lithotrophically on gases present on early Earth. Among the organic molecules present on early Earth is acetaldehyde, a highly volatile C2 compound. Here, we demonstrate that the acetogenic model bacterium Acetobacterium woodii grows on acetaldehyde. Acetaldehyde is dismutated to ethanol and acetyl-CoA, most likely by the bifunctional alcohol dehydrogenase AdhE. Acetyl-CoA is converted to acetate by two subsequent enzymes, phosphotransacetylase and acetate kinase, accompanied by the synthesis of ATP by substrate-level phosphorylation. Apparently, growth on acetaldehyde does not employ the Wood-Ljungdahl pathway. Our finding opens the possibility of a simple and ancient metabolic pathway with only three enzymes that allows for biomass (acetyl-CoA) and ATP formation on early Earth.

Effects of short light/dark cycles on photosynthetic pathway switching and growth of medicinal Dendrobium officinale in aeroponic cultivation

Dendrobium officinale has high medicinal value but grows slowly in natural environment due to its special CAM photosynthetic pathway. In this study, D. officinale were grown aeroponically with light/dark cycles of 12 h/12 h, 4 h/4 h, and 2 h/2 h for 150 d. The photosynthetic electron transfer characteristics, photosynthetic CO2 fixation pathways, and accumulations of biomass and soluble polysaccharides in D. officinale leaves were studied. The results showed that the photosynthetic apparatus states of D. officinale in aeroponic cultivation under short light/dark cycles of 4 h/4 h and 2 h/2 h were better than that under 12 h/12 h. The dark net CO2 exchange percentages of D. officinale were negative in short light/dark cycles of 4 h/4 h and 2 h/2 h, and the daily net CO2 exchange amount and dry/fresh weight increases were doubled compared with those in 12 h/12 h light/dark cycle. High soluble polysaccharides content and the soluble polysaccharides yield of D. officinale were obtained in the shorter light/dark cycle of 2 h/2 h. Therefore, the photosynthetic pathway of D. officinale could be switched from CAM to C3 by short light/dark cycles of 4 h/4 h and 2 h/2 h, and its higher biomass accumulation and soluble polysaccharides yield could be obtained by the shorter light/dark cycle of 2 h/2 h in aeroponic cultivation. Keywords: dark net CO2 exchange percentage, photosynthetic pathway, short light/dark cycle, soluble polysaccharides DOI: 10.25165/j.ijabe.20191205.4864 Citation: Cheng Y S, He D X, He J, Niu G H, Ji F. Effects of short light/dark cycles on photosynthetic pathway switching and growth of medicinal Dendrobium officinale in aeroponic cultivation. Int J Agric & Biol Eng, 2019; 12(5): 38–43.

Form III RubisCO-mediated transaldolase variant of the Calvin cycle in a chemolithoautotrophic bacterium

The Calvin-Benson-Bassham (CBB) cycle assimilates CO2 for the primary production of organic matter in all plants and algae, as well as in some autotrophic bacteria. The key enzyme of the CBB cycle, ribulose-bisphosphate carboxylase/oxygenase (RubisCO), is a main determinant of de novo organic matter production on Earth. Of the three carboxylating forms of RubisCO, forms I and II participate in autotrophy, and form III so far has been associated only with nucleotide and nucleoside metabolism. Here, we report that form III RubisCO functions in the CBB cycle in the thermophilic chemolithoautotrophic bacterium Thermodesulfobium acidiphilum, a phylum-level lineage representative. We further show that autotrophic CO2 fixation in T. acidiphilum is accomplished via the transaldolase variant of the CBB cycle, which has not been previously demonstrated experimentally and has been considered unlikely to occur. Thus, this work reveals a distinct form of the key pathway of CO2 fixation.

Advances in metabolic engineering of heterotrophic microorganisms for CO₂ fixation: a review

Environmental protection and energy supply are our two major concerns. Greenhouse gases released from energy consumption have serious impact on the environment. CO₂ fixation can be used to convert CO₂ into fuels or chemicals. However, natural carbon-fixing organisms usually have some disadvantages such as slow growth and low carbon fixation efficiency. Enhancing or remodeling CO₂ fixation pathways in model microorganisms can realize CO₂ recycling, which can further increase fuel or chemical production and reduce greenhouse gas emission. This review describes in detail metabolic engineering of CO₂ fixation pathways to improve chemical production and sugar synthesis, elaborates the role of relevant metabolic pathways and key enzymes in CO₂ fixation, introduces the application of electro-biochemical synthesis system, shows the great potential of CO₂ fixation, and prospects the future research direction of CO₂ fixation.

Shifts in microbial community and carbon sequestration in farmland soil under long-term conservation tillage and straw returning

Soil microorganisms participate in soil carbon cycling and they are affected by tillage and straw returning. In this experiment, a split-plot design was adopted beginning in 2002. The whole-plot treatment had three tillage modes, conventional tillage (CT), subsoil tillage (ST) and rotary tillage (RT), and the split-plot treatment had two levels of maize straw returning, full straw returning (F) and no straw returning (0). The soil organic carbon (SOC) contents during 2012–2017 were measured, whereas the microbial community function and structure were assessed by Biolog EcoPlates® and Illumina sequencing technology. The results showed that the SOC increased under all treatments from 2012 to 2017. However, the growth rate for SOC content under STF was the highest. Subsoil tillage and full straw returning significantly increased the metabolic function and metabolic diversity of the soil microbial carbon sources. A principal component analysis (PCA) showed that except for CT0 and RT0, the microbial metabolic functions under the various treatments were different. Amino acids and amines were the main carbon sources that caused changes in the soil microbial communities. A redundancy analysis (RDA) showed that the soil water content, bulk density, total nitrogen (TN) content, SOC and pH significantly accounted for the variance in the microbial community function and structure; however, the bulk density was not conducive to variation in the microbial communities. Subsoil tillage with full straw returning could increase the gene abundances in the metabolic pathways for CO2 fixation and organic acids but decrease the gene abundance in the di- and oligosaccharide metabolic pathway. In conclusion, a combination of subsoil tillage and full straw returning improve the basic soil physicochemical and biological properties, affect soil microbial communities, and promote an increase in the SOC content, which can eventually resolve soil degradation issues.

Effect of Phosphorus Addition on the Abundance of Autotrophic CO2-Fixation Microorganisms in Rhizospheric Soil from a Phosphorus-Limited Paddy Field

A rice pot experiment was conducted to investigate the effect of phosphorus addition on the abundance of autotrophic CO2-fixation microorganisms using phosphorus-limited paddy soil from the Changsha Observation and Research Station for the Agricultural Environment. Rice seedlings were transplanted in the paddy soil with or without phosphorus addition, corresponding to P-treated-pot (P) or control pot (CK), respectively. Rhizosphere soils were collected from the P and CK treatments during the tillering and shooting stages. The physical and chemical soil properties were measured and the abundance of autotrophic CO2-fixation microorganisms was quantified with a real-time PCR technique based on four functional genes (cbbL, cbbM, accA, and aclB) involved in three CO2-fixation pathways (CBB cycle, rTCA cycle, and 3-hydroxypropionate/4-hydroxybutyrate cycle). The results show that phosphorus addition improves the concentrations of DOC and Olsen-P and the pH value, whereas negative effects on the MBC and NH4+-N concentrations are revealed during the tillering stage. The effect of phosphorus addition on the NO3--N concentration in the tillering and shooting stages differs. Phosphorus addition significantly increases the abundances of the cbbL, cbbM, accA, and aclB genes, which are 156%, 99%, 110%, and 193% higher than those of the CK treatment in the tillering stage. However, this positive effect is not notable for the cbbL, accA, and aclB genes during the shooting stage. Redundancy analysis (RDA) shows that Olsen-P is the environmental factor that most significantly affects the abundance of autotrophic CO2-fixation microorganisms.

Photoreduction of CO2 with a Formate Dehydrogenase Driven by Photosystem II Using a Semi-artificial Z-Scheme Architecture.

Solar-driven coupling of water oxidation with CO2 reduction sustains life on our planet and is of high priority in contemporary energy research. Here, we report a photoelectrochemical tandem device that performs photocatalytic reduction of CO2 to formate. We employ a semi-artificial design, which wires a W-dependent formate dehydrogenase (FDH) cathode to a photoanode containing the photosynthetic water oxidation enzyme, Photosystem II, via a synthetic dye with complementary light absorption. From a biological perspective, the system achieves a metabolically inaccessible pathway of light-driven CO2 fixation to formate. From a synthetic point of view, it represents a proof-of-principle system utilizing precious-metal-free catalysts for selective CO2-to-formate conversion using water as an electron donor. This hybrid platform demonstrates the translatability and versatility of coupling abiotic and biotic components to create challenging models for solar fuel and chemical synthesis.

Microbial activity response to hydrogen injection in thermophilic anaerobic digesters revealed by genome-centric metatranscriptomics

BackgroundThe expansion of renewable energy produced by windmills and photovoltaic panels has generated a considerable electricity surplus, which can be utilized in water electrolysis systems for hydrogen production. The resulting hydrogen can then be funneled to anaerobic digesters for biogas upgrading (biomethanation) purposes (power-to-methane) or to produce high value-added compounds such as short-chain fatty acids (power-to-chemicals).Genome-centric metagenomics and metatranscriptomic analyses were performed to better understand the metabolic dynamics associated with H2 injection in two different configurations of anaerobic digesters treating acidic wastes, specifically cheese manufacturing byproducts. These approaches revealed the key-genes involved in methanation and carbon fixation pathways at species level.ResultsThe biogas upgrading process in the single-stage configuration increased the CH4 content by 7%. The dominant methanogenic species responsible for the upregulation of the hydrogenotrophic pathway in this reactor was Methanothermobacter wolfeii UC0008. In the two-stage configuration, H2 injection induced an upregulation of CO2 fixation pathways producing short-chain fatty acids, mainly acetate and butyrate. In this configuration, the abundant species Anaerobaculum hydrogeniformans UC0046 and Defluviitoga tunisiensis UC0050 primarily upregulated genes related to electron transport chains, suggesting putative syntrophisms with hydrogen scavenger microbes. Interestingly, Tepidanaerobacter acetatoxydans UC0018 did not act as an acetate-oxidizer in either reactor configurations, and instead regulated pathways involved in acetate production and uptake. A putative syntrophic association between Coprothermobacter proteolyticus UC0011 and M. wolfeii UC0008 was proposed in the two-stage reactor. In order to support the transcriptomic findings regarding the hydrogen utilization routes, an advanced bioconversion model was adapted for the simulation of the single- and two-stage reactor setups.ConclusionsThis is the first study investigating biogas reactor metatranscriptome dynamics following hydrogen injection for biomethanation and carbon fixation to short-chain fatty acids purposes. The same microbes showed different patterns of metabolic regulation in the two reactor configurations. It was observed an effect of the specialized acidogenic reactor on the overall microbial consortium composition and activity in the two-stage digester. There were also suggested the main species responsible for methanation, short-chain fatty acids production, and electron transport chain mechanisms, in both reactor configurations.

Diurnal Changes in Active Carbon and Nitrogen Pathways Along the Temperature Gradient in Porcelana Hot Spring Microbial Mat.

Composition, carbon and nitrogen uptake, and gene transcription of microbial mat communities in Porcelana neutral hot spring (Northern Chilean Patagonia) were analyzed using metagenomics, metatranscriptomics and isotopically labeled carbon (H13CO3) and nitrogen (15NH4Cl and K15NO3) assimilation rates. The microbial mat community included 31 phyla, of which only Cyanobacteria and Chloroflexi were dominant. At 58°C both phyla co-occurred, with similar contributions in relative abundances in metagenomes and total transcriptional activity. At 66°C, filamentous anoxygenic phototrophic Chloroflexi were >90% responsible for the total transcriptional activity recovered, while Cyanobacteria contributed most metagenomics and metatranscriptomics reads at 48°C. According to such reads, phototrophy was carried out both through oxygenic photosynthesis by Cyanobacteria (mostly Mastigocladus) and anoxygenic phototrophy due mainly to Chloroflexi. Inorganic carbon assimilation through the Calvin–Benson cycle was almost exclusively due to Mastigocladus, which was the main primary producer at lower temperatures. Two other CO2 fixation pathways were active at certain times and temperatures as indicated by transcripts: 3-hydroxypropionate (3-HP) bi-cycle due to Chloroflexi and 3-hydroxypropionate-4-hydroxybutyrate (HH) cycle carried out by Thaumarchaeota. The active transcription of the genes involved in these C-fixation pathways correlated with high in situ determined carbon fixation rates. In situ measurements of ammonia assimilation and nitrogen fixation (exclusively attributed to Cyanobacteria and mostly to Mastigocladus sp.) showed these were the most important nitrogen acquisition pathways at 58 and 48°C. At 66°C ammonia oxidation genes were actively transcribed (mostly due to Thaumarchaeota). Reads indicated that denitrification was present as a nitrogen sink at all temperatures and that dissimilatory nitrate reduction to ammonia (DNRA) contributed very little. The combination of metagenomic and metatranscriptomic analysis with in situ assimilation rates, allowed the reconstruction of day and night carbon and nitrogen assimilation pathways together with the contribution of keystone microorganisms in this natural hot spring microbial mat.

OptMDFpathway: Identification of metabolic pathways with maximal thermodynamic driving force and its application for analyzing the endogenous CO2 fixation potential of Escherichia coli.

Constraint-based modeling techniques have become a standard tool for the in silico analysis of metabolic networks. To further improve their accuracy, recent methodological developments focused on integration of thermodynamic information in metabolic models to assess the feasibility of flux distributions by thermodynamic driving forces. Here we present OptMDFpathway, a method that extends the recently proposed framework of Max-min Driving Force (MDF) for thermodynamic pathway analysis. Given a metabolic network model, OptMDFpathway identifies both the optimal MDF for a desired phenotypic behavior as well as the respective pathway itself that supports the optimal driving force. OptMDFpathway is formulated as a mixed-integer linear program and is applicable to genome-scale metabolic networks. As an important theoretical result, we also show that there exists always at least one elementary mode in the network that reaches the maximal MDF. We employed our new approach to systematically identify all substrate-product combinations in Escherichia coli where product synthesis allows for concomitant net CO2 assimilation via thermodynamically feasible pathways. Although biomass synthesis cannot be coupled to net CO2 fixation in E. coli we found that as many as 145 of the 949 cytosolic carbon metabolites contained in the genome-scale model iJO1366 enable net CO2 incorporation along thermodynamically feasible pathways with glycerol as substrate and 34 with glucose. The most promising products in terms of carbon assimilation yield and thermodynamic driving forces are orotate, aspartate and the C4-metabolites of the tricarboxylic acid cycle. We also identified thermodynamic bottlenecks frequently limiting the maximal driving force of the CO2-fixing pathways. Our results indicate that heterotrophic organisms like E. coli hold a possibly underestimated potential for CO2 assimilation which may complement existing biotechnological approaches for capturing CO2. Furthermore, we envision that the developed OptMDFpathway approach can be used for many other applications within the framework of constrained-based modeling and for rational design of metabolic networks.

Serpentinization: Connecting Geochemistry, Ancient Metabolism and Industrial Hydrogenation.

Rock–water–carbon interactions germane to serpentinization in hydrothermal vents have occurred for over 4 billion years, ever since there was liquid water on Earth. Serpentinization converts iron(II) containing minerals and water to magnetite (Fe3O4) plus H2. The hydrogen can generate native metals such as awaruite (Ni3Fe), a common serpentinization product. Awaruite catalyzes the synthesis of methane from H2 and CO2 under hydrothermal conditions. Native iron and nickel catalyze the synthesis of formate, methanol, acetate, and pyruvate—intermediates of the acetyl-CoA pathway, the most ancient pathway of CO2 fixation. Carbon monoxide dehydrogenase (CODH) is central to the pathway and employs Ni0 in its catalytic mechanism. CODH has been conserved during 4 billion years of evolution as a relic of the natural CO2-reducing catalyst at the onset of biochemistry. The carbide-containing active site of nitrogenase—the only enzyme on Earth that reduces N2—is probably also a relic, a biological reconstruction of the naturally occurring inorganic catalyst that generated primordial organic nitrogen. Serpentinization generates Fe3O4 and H2, the catalyst and reductant for industrial CO2 hydrogenation and for N2 reduction via the Haber–Bosch process. In both industrial processes, an Fe3O4 catalyst is matured via H2-dependent reduction to generate Fe5C2 and Fe2N respectively. Whether serpentinization entails similar catalyst maturation is not known. We suggest that at the onset of life, essential reactions leading to reduced carbon and reduced nitrogen occurred with catalysts that were synthesized during the serpentinization process, connecting the chemistry of life and Earth to industrial chemistry in unexpected ways.

Investigating the Composition and Metabolic Potential of Microbial Communities in Chocolate Pots Hot Springs.

Iron (Fe) redox-based metabolisms likely supported life on early Earth and may support life on other Fe-rich rocky planets such as Mars. Modern systems that support active Fe redox cycling such as Chocolate Pots (CP) hot springs provide insight into how life could have functioned in such environments. Previous research demonstrated that Fe- and Si-rich and slightly acidic to circumneutral-pH springs at CP host active dissimilatory Fe(III) reducing microorganisms. However, the abundance and distribution of Fe(III)-reducing communities at CP is not well-understood, especially as they exist in situ. In addition, the potential for direct Fe(II) oxidation by lithotrophs in CP springs is understudied, in particular when compared to indirect oxidation promoted by oxygen producing Cyanobacteria. Here, a culture-independent approach, including 16S rRNA gene amplicon and shotgun metagenomic sequencing, was used to determine the distribution of putative Fe cycling microorganisms in vent fluids and sediment cores collected along the outflow channel of CP. Metagenome-assembled genomes (MAGs) of organisms native to sediment and planktonic microbial communities were screened for extracellular electron transfer (EET) systems putatively involved in Fe redox cycling and for CO2 fixation pathways. Abundant MAGs containing putative EET systems were identified as part of the sediment community at locations where Fe(III) reduction activity has previously been documented. MAGs encoding both putative EET systems and CO2 fixation pathways, inferred to be FeOB, were also present, but were less abundant components of the communities. These results suggest that the majority of the Fe(III) oxides that support in situ Fe(III) reduction are derived from abiotic oxidation. This study provides new insights into the interplay between Fe redox cycling and CO2 fixation in sustaining chemotrophic communities in CP with attendant implications for other neutral-pH hot springs.