Biological Conversion of Formate to Organic Compounds: Toward a Sustainable Formate Bioeconomy

Abstract

Formate bioconversion plays a crucial role in achieving renewable resource utilization and green and sustainable development, as it helps convert formate to biofuels and biochemicals. However, to tap the full potential of formate bioconversion, it is important to identify the most appropriate microbial hosts, design the most promising formate assimilation pathways, and develop the most efficient formate assimilation cell factories. Here, we summarize the formatotrophic microorganisms capable of assimilating formate into building blocks of cell growth and analyze the characteristics of formate assimilation pathways for transmitting formate into central carbon metabolism. Furthermore, we discuss microbial engineering strategies to improve the efficiency of formate utilization for producing high-value bioproducts. Finally, we highlight the key challenges of formate bioconversion and their possible solutions to advance the formate bioeconomy and biomanufacturing.

Keywords

formate / formate assimilation pathways / formatotrophic microorganisms / microbial engineering

References

[1] S. J. Kim, J. Yoon, D. K. Im, Y. H. Kim, and M. K. Oh, “Adaptively Evolved Escherichia coli for Improved Ability of Formate Utilization as a Carbon Source in Sugar-Free Conditions,” Biotechnology for Biofuels 12, no. 1 (2019): 207.

[2] K. Wang, Y. Da, H. Bi, et al., “A One-Carbon Chemicals Conversion Strategy to Produce Precursor of Biofuels With Saccharomyces cerevisiae,” Renewable Energy 208 (2023): 331–340.

[3] L. Calzadiaz-Ramirez and A. S. Meyer, “Formate Dehydrogenases for CO₂ Utilization,” Current Opinion in Biotechnology 73 (2022): 95–100.

[4] W. Zhong, H. Li, and Y. Wang, “Design and Construction of Artificial Biological Systems for One-Carbon Utilization,” Biodesign Research 5 (2023): 0021.

[5] L. R. Dahlin, A. W. Meyers, S. W. Stefani, et al., “Heterologous Expression of Formate Dehydrogenase Enables Photoformatotrophy in the Emerging Model Microalga, Picochlorum renovo,” Frontiers in Bioengineering and Biotechnology 11 (2023): 1162745.

[6] I. Ergal, B. Reischl, B. Hasibar, et al., “Formate Utilization by the Crenarchaeon Desulfurococcus amylolyticus,” Microorganisms 8, no. 3 (2020): 454.

[7] Q. Chen, Y. Chen, Z. Hou, et al., “Unlocking the Formate Utilization of Wild-Type Yarrowia lipolytica Through Adaptive Laboratory Evolution,” Biotechnology Journal 19, no. 6 (2024): e2400290.

[8] M. Moon, G. W. Park, J. P. Lee, J. S. Lee, and K. Min, “Recombinant Expression and Characterization of Formate Dehydrogenase From Clostridium ljungdahlii (CLFDH) as CO₂ Reductase for Converting CO₂ to Formate,” Journal of CO2 Utilization 57 (2022): 101876.

[9] K. Min, M. Moon, G. W. Park, J. P. Lee, S. J. Kim, and J. S. Lee, “Newly Explored Formate Dehydrogenases From Clostridium Species Catalyze Carbon Dioxide to Formate,” Bioresource Technology 348 (2022): 126832.

[10] T. J. Wang, L. B. Sun, X. Ai, P. Chen, Y. Chen, and X. Wang, “Boosting Formate Electrooxidation by Heterostructured PtPd Alloy and Oxides Nanowires,” Advanced Materials 36, no. 27 (2024): e2403664.

[11] Y. W. Zhou, Y. F. Chen, X. Qin, K. Jiang, W. F. Lin, and W. B. Cai, “Boosting Electrocatalytic Oxidation of Formic Acid on SnO₂-Decorated Pd Nanosheets,” Journal of Catalysis 399 (2021): 8–14.

[12] O. Yishai, L. Goldbach, H. Tenenboim, S. N. Lindner, and A. Bar-Even, “Engineered Assimilation of Exogenous and Endogenous Formate in Escherichia coli,” ACS Synthetic Biology 6, no. 9 (2017): 1722–1731.

[13] Y. Yao, B. Fu, D. Han, Y. Zhang, and H. Liu, “Formate-Dependent Acetogenic Utilization of Glucose by the Fecal Acetogen Clostridium bovifaecis,” Applied and Environmental Microbiology 86, no. 23 (2020): e01870-20, https://doi.org/10.1128/AEM.01870-20.

[14] V. R. Bysani, A. S. Alam, A. Bar-Even, and F. Machens, “Engineering and Evolution of the Complete Reductive Glycine Pathway in Saccharomyces cerevisiae for Formate and CO₂ Assimilation,” Metabolic Engineering 81 (2024): 167–181.

[15] N. G, I. A. K, and Z. Z, “Experimental Validation of In Silico Flux Predictions From a Genome-Scale Model (IMM518) for Carbon Dioxide Utilization by M. maripaludis,” Computer Aided Chemical Engineering 37 (2015): 2153–2158.

[16] R. Zhao, W. Dong, C. Yang, W. Jiang, J. Tian, and Y. Gu, “Formate as a Supplementary Substrate Facilitates Sugar Metabolism and Solvent Production by Clostridium beijerinckii NCIMB 8052,” Synthetic and Systems Biotechnology 8, no. 2 (2023): 196–205.

[17] J. R. Humphreys, S. D. Hebdon, H. Rohrer, et al., “Establishing Butyribacterium methylotrophicum as a Platform Organism for the Production of Biocommodities From Liquid C1 Metabolites,” Applied and Environmental Microbiology 88, no. 6 (2022): e0239321.

[18] S. Kim, S. N. Lindner, S. Aslan, et al., “Growth of E. coli on Formate and Methanol via the Reductive Glycine Pathway,” Nature Chemical Biology 16, no. 5 (2020): 538–545.

[19] J. B. Siegel, A. L. Smith, S. Poust, et al., “Computational Protein Design Enables a Novel One-Carbon Assimilation Pathway,” Proceedings of the National Academy of Sciences 112, no. 12 (2015): 3704–3709.

[20] A. Bar-Even, “Formate Assimilation: The Metabolic Architecture of Natural and Synthetic Pathways,” Biochemistry 55, no. 28 (2016): 3851–3863.

[21] W. Mao, Q. Yuan, H. Qi, Z. Wang, H. Ma, and T. Chen, “Recent Progress in Metabolic Engineering of Microbial Formate Assimilation,” Applied Microbiology and Biotechnology 104, no. 16 (2020): 6905–6917.

[22] C. A. Cotton, N. J. Claassens, S. Benito-Vaquerizo, and A. Bar-Even, “Renewable Methanol and Formate as Microbial Feedstocks,” Current Opinion in Biotechnology 62 (2020): 168–180.

[23] S. Bierbaumer, M. Nattermann, L. Schulz, et al., “Enzymatic Conversion of CO(2): From Natural to Artificial Utilization,” Chemical Reviews 123, no. 9 (2023): 5702–5754.

[24] C. S. Neuendorf, G. A. Vignolle, C. Derntl, et al., “A Quantitative Metabolic Analysis Reveals Acetobacterium woodii as a Flexible and Robust Host for Formate-Based Bioproduction,” Metabolic Engineering 68 (2021): 68–85.

[25] S. Kim, S. H. Lee, H. Seo, and K. J. Kim, “Biochemical Properties and Crystal Structure of Formate-Tetrahydrofolate Ligase From Methylobacterium extorquens CM4,” Biochemical and Biophysical Research Communications 528, no. 3 (2020): 426–431.

[26] S. M. da Silva, J. Voordouw, C. Leitão, M. Martins, G. Voordouw, and I. Pereira, “Function of Formate Dehydrogenases in Desulfovibrio vulgaris Hildenborough Energy Metabolism,” Microbiology 159 (2013): 1760–1769.

[27] Y. Tashiro, S. Hirano, M. M. Matson, S. Atsumi, and A. Kondo, “Electrical-Biological Hybrid System for CO₂ Reduction,” Metabolic Engineering 47 (2018): 211–218.

[28] S. Gleizer, R. Ben-Nissan, Y. M. Bar-On, et al., “Conversion of Escherichia coli to Generate All Biomass Carbon From CO₂,” Cell 179, no. 6 (2019): 1255–1263.e12.

[29] J. Bang and S. Y. Lee, “Assimilation of Formic Acid and CO₂ by Engineered Escherichia coli Equipped With Reconstructed One-Carbon Assimilation Pathways,” Proceedings of the National Academy of Sciences of the United States of America 115, no. 40 (2018): 9271.

[30] J. Xu, Engineering Formate Assimilation in E. coli Using Stable Isotope Tracers and Thermodynamic Analysis (UCLA, 2020).

[31] J. Gonzalez de la Cruz, F. Machens, K. Messerschmidt, and A. Bar-Even, “Core Catalysis of the Reductive Glycine Pathway Demonstrated in Yeast,” ACS Synthetic Biology 8, no. 5 (2019): 911–917.

[32] A. C. A. van Aalst, R. Mans, and J. T. Pronk, “An Engineered Non-Oxidative Glycolytic Bypass Based on Calvin-Cycle Enzymes Enables Anaerobic Co-Fermentation of Glucose and Sorbitol by Saccharomyces cerevisiae,” Biotechnology for Biofuels and Bioproducts 15, no. 1 (2022): 112.

[33] B. M. Mitic, C. Troyer, L. Lutz, M. Baumschabl, S. Hann, and D. Mattanovich, “The Oxygen-Tolerant Reductive Glycine Pathway Assimilates Methanol, Formate and CO₂ in the Yeast Komagataella phaffii,” Nature Communications 14, no. 1 (2023): 7754.

[34] J. Turlin, B. Dronsella, A. De Maria, S. N. Lindner, and P. I. Nikel, “Integrated Rational and Evolutionary Engineering of Genome-Reduced Pseudomonas putida Strains Promotes Synthetic Formate Assimilation,” Metabolic Engineering 74 (2022): 191–205.

[35] K. Li, X. Zhang, C. Li, et al., “Systems Metabolic Engineering of Corynebacterium glutamicum to Assimilate Formic Acid for Biomass Accumulation and Succinic Acid Production,” Bioresource Technology 402 (2024): 130774.

[36] M. Straub, M. Demler, D. Weuster-Botz, and P. Dürre, “Selective Enhancement of Autotrophic Acetate Production With Genetically Modified Acetobacterium woodii,” Journal of Biotechnology 178 (2014): 67–72.

[37] Y. Yao, B. Fu, D. Han, Y. Zhang, Z. Wei, and H. Liu, “Reduction, Evolutionary Pattern and Positive Selection of Genes Encoding Formate Dehydrogenase in Wood-Ljungdahl Pathway of Gastrointestinal Acetogens Suggests Their Adaptation to Formate-Rich Habitats,” Environmental Microbiology Reports 15, no. 2 (2023): 129–141.

[38] F. M. Schwarz, S. Ciurus, S. Jain, et al., “Revealing Formate Production From Carbon Monoxide in Wild Type and Mutants of Rnf- and Ech-Containing Acetogens, Acetobacterium woodii and Thermoanaerobacter kivui,” Microbial Biotechnology 13, no. 6 (2020): 2044–2056.

[39] S. Grunwald, A. Mottet, E. Grousseau, et al., “Kinetic and Stoichiometric Characterization of Organoautotrophic Growth of Ralstonia eutropha on Formic Acid in Fed-Batch and Continuous Cultures,” Microbial Biotechnology 8, no. 1 (2015): 155–163.

[40] J. Moon, J. Dönig, S. Kramer, A. Poehlein, R. Daniel, and V. Müller, “Formate Metabolism in the Acetogenic Bacterium Acetobacterium woodii,” Environmental Microbiology 23, no. 8 (2021): 4214–4227.

[41] R. Conrad, C. Erkel, and W. Liesack, “Rice Cluster I Methanogens, An Important Group of Archaea Producing Greenhouse Gas in Soil,” Current Opinion in Biotechnology 17, no. 3 (2006): 262–267.

[42] Z. Lü and Y. Lu, “Methanocella conradii sp. nov., a Thermophilic, Obligate Hydrogenotrophic Methanogen, Isolated From Chinese Rice Field Soil,” PLoS One 7, no. 4 (2012): e35279.

[43] M. F. Abdul Halim, L. A. Day, and K. C. Costa, “Formate-Dependent Heterodisulfide Reduction in a Methanomicrobiales Archaeon,” Applied and Environmental Microbiology 87, no. 6 (2021): e02698-02620.

[44] S. P. Gilmore, J. K. Henske, J. A. Sexton, et al., “Genomic Analysis of Methanogenic Archaea Reveals a Shift Towards Energy Conservation,” BMC Genomics 18 (2017): 639.

[45] Y. Liu and W. B. Whitman, “Metabolic, Phylogenetic, and Ecological Diversity of the Methanogenic Archaea,” Annals of the New York Academy of Sciences 1125, no. 1 (2008): 171–189.

[46] N. Matschiavelli and M. Rother, “Role of a Putative Tungsten-Dependent Formylmethanofuran Dehydrogenase in Methanosarcina acetivorans,” Archives of Microbiology 197, no. 3 (2015): 379–388.

[47] C. B. Walker, A. M. Redding-Johanson, E. E. Baidoo, et al., “Functional Responses of Methanogenic Archaea to Syntrophic Growth,” ISME Journal 6, no. 11 (2012): 2045–2055.

[48] W. M. Chen, C. Y. Cai, Z. H. Li, C. C. Young, and S. Y. Sheu, “Methylobacterium oryzihabitans sp. nov., isolated From Water Sampled From a Rice Paddy Field,” International Journal of Systematic and Evolutionary Microbiology 69, no. 12 (2019): 3843–3850.

[49] P. Jourand, E. Giraud, G. Béna, et al., “Methylobacterium nodulans sp nov., for a Group of Aerobic, Facultatively Methylotrophic, Legume Root-Nodule-Forming and Nitrogen-Fixing Bacteria,” International Journal of Systematic and Evolutionary Microbiology 54 (2004): 2269–2273.

[50] R. P. Singh, R. N. Singh, M. K. Srivastava, et al., “Structure Prediction and Analysis of MxaF From Obligate, Facultative and Restricted Facultative Methylobacterium,” Bioinformation 8, no. 21 (2012): 1042–1046.

[51] G. J. Crowther, G. Kosály, and M. E. Lidstrom, “Formate as the Main Branch Point for Methylotrophic Metabolism in Methylobacterium extorquens Am1,” Journal of Bacteriology 190, no. 14 (2008): 5057–5062.

[52] L. Y. Cui, J. Yang, W. F. Liang, S. Yang, C. Zhang, and X. Xing, “Sodium Formate Redirects Carbon Flux and Enhances Heterologous Mevalonate Production in Methylobacterium extorquens Am1,” Biotechnology Journal 18, no. 2 (2023): e2200402.

[53] J. Park, Y. Heo, B. W. Jeon, et al., “Structure of Recombinant Formate Dehydrogenase From Methylobacterium extorquens (MeFDH1),” Scientific Reports 14, no. 1 (2024): 3819.

[54] L. Chistoserdova, M. Laukel, J. C. Portais, J. A. Vorholt, and M. E. Lidstrom, “Multiple Formate Dehydrogenase Enzymes in the Facultative Methylotroph Methylobacterium extorquens AM1 Are Dispensable for Growth on Methanol,” Journal of Bacteriology 186, no. 1 (2004): 22–28.

[55] R. Zhao and J. F. Biddle, “Helarchaeota and Co-Occurring Sulfate-Reducing Bacteria in Subseafloor Sediments From the Costa Rica Margin,” Isme Communications 1, no. 1 (2021): 25.

[56] M. Roy. “The Effects of Electron Donors on the Growth of Sulfate-Reducing Bacteria in Copper-Zinc and Gold Mine Tailings From Timmins, Ontario,” Masters Abstracts International, 43-06: 2144 (2004).

[57] S. Baena, M. L. Fardeau, M. Labat, B. Ollivier, J. L. Garcia, and B. Patel, “Desulfovibrio aminophilus Sp. Nov., a Novel Amino Acid Degrading and Sulfate Reducing Bacterium From An Anaerobic Dairy Wastewater Lagoon,” Systematic and Applied Microbiology 21, no. 4 (1998): 498–504.

[58] B. H. G. W. van Houten, R. J. W. Meulepas, W. van Doesburg, H. Smidt, G. Muyzer, and A. Stams, “Desulfovibrio paquesii Sp. Nov., a Hydrogenotrophic Sulfate-Reducing Bacterium Isolated From a Synthesis-Gas-Fed Bioreactor Treating Zinc- and Sulfate-Rich Wastewater,” International Journal of Systematic and Evolutionary Microbiology 59 (2009): 229–233.

[59] M. N. Price, J. Ray, K. M. Wetmore, et al., “The Genetic Basis of Energy Conservation in the Sulfate-Reducing Bacterium Desulfovibrio alaskensis G20,” Frontiers in Microbiology 5 (2014): 577.

[60] I. Sánchez-Andrea, I. A. Guedes, B. Hornung, et al., “The Reductive Glycine Pathway Allows Autotrophic Growth of Desulfovibrio desulfuricans,” Nature Communications 11, no. 1 (2020): 5090.

[61] Y. Liu, J. Vaughan, G. Southam, et al., “Role of the Substrate on Ni Inhibition in Biological Sulfate Reduction,” Journal of Environmental Management 316 (2022): 115216.

[62] S. Tong, L. Zhao, D. Zhu, W. Chen, L. Chen, and D. Li, “From Formic Acid to Single-Cell Protein: Genome-Scale Revealing the Metabolic Network of Paracoccus communis Ma5,” Bioresources and Bioprocessing 9, no. 1 (2022): 55.

[63] M. Janasch, N. Crang, J. Asplund-Samuelsson, et al., “Thermodynamic Limitations of PHB Production From Formate and Fructose in Cupriavidus necator,” Metabolic Engineering 73 (2022): 256–269.

[64] B. S. Pickering and I. J. Oresnik, “Formate-Dependent Autotrophic Growth in Sinorhizobium meliloti,” Journal of Bacteriology 190, no. 19 (2008): 6409–6418.

[65] C. E. Lawson, A. B. Mundinger, H. Koch, et al., “Investigating the Chemolithoautotrophic and Formate Metabolism of Nitrospira moscoviensis by Constraint-Based Metabolic Modeling and 13C-tracer Analysis,” mSystems 6, no. 4 (2021): e0017321, https://doi.org/10.1128/mSystems.00173-21.

[66] J. Li, L. Zhang, Q. Xu, et al., “CRISPR-Cas9 Toolkit for Genome Editing iIn an Autotrophic CO₂-Fixing Methanogenic Archaeon,” Microbiology Spectrum 10, no. 4 (2022): e0116522.

[67] T. Jiang, C. Zhang, Q. He, Z. Zheng, and J. Ouyang, “Metabolic Engineering of Escherichia coli K12 for Homofermentative Production of L-Lactate From Xylose,” Applied Biochemistry and Biotechnology 184, no. 2 (2018): 703–715.

[68] C. A. Cotton, C. Edlich-Muth, and A. Bar-Even, “Reinforcing Carbon Fixation: CO₂ Reduction Replacing and Supporting Carboxylation,” Current Opinion in Biotechnology 49 (2018): 49–56.

[69] V. F. K. Yuen, D. T. Z. Jun, and K. Zhou, “Improving Glycine Utilization in Escherichia coli,” Biochemical Engineering Journal 193 (2023): 108834.

[70] O. Yishai, M. Bouzon, V. Döring, and A. Bar-Even, “In Vivo Assimilation of One-Carbon via a Synthetic Reductive Glycine Pathway in Escherichia coli,” ACS Synthetic Biology 7, no. 9 (2018): 2023–2028.

[71] S. Zobel, J. Kuepper, B. Ebert, N. Wierckx, and L. M. Blank, “Metabolic Response of Pseudomonas putida to Increased NADH Regeneration Rates,” Engineering in Life Sciences 17, no. 1 (2017): 47–57.

[72] A. Roca, J. J. Rodríguez-Herva, and J. L. Ramos, “Redundancy of Enzymes for Formaldehyde Detoxification in Pseudomonas putida,” Journal of Bacteriology 191, no. 10 (2009): 3367–3374.

[73] V. Riis, D. Miethe, and W. Babel, “Formate-Stimulated Oxidation of Methanol by Pseudomonas putida 9816,” Bioscience, Biotechnology, and Biochemistry 67, no. 4 (2003): 684–690.

[74] O. V. Tyurin and D. G. Kozlov, “Deletion of the Fld Gene in Methylotrophic Yeasts Komagataella phaffii and Komagataella kurtzmanii Results in Enhanced Induction of the AOX1 Promoter in Response to Either Methanol or Formate,” Microbiology 84, no. 3 (2015): 408–411.

[75] B. Liu, H. Li, H. Zhou, and J. Zhang, “Enhancing Xylanase Expression by Komagataella phaffii by Formate as Carbon Source and Inducer,” Applied Microbiology and Biotechnology 106, no. 23 (2022): 7819–7829.

[76] F. M. Kashiwagi, Y. Ojima, and M. Taya, “Metabolic Engineering of Escherichia coli KO11 With the NADH Regeneration System for Enhancing Ethanol Production,” Journal of Chemical Engineering of Japan 51, no. 3 (2018): 264–268.

[77] H. Yu and J. C. Liao, “A Modified Serine Cycle in Escherichia coli Coverts Methanol and CO₂ to Two-Carbon Compounds,” Nature Communications 9 (2018): 3992.

[78] X. Zhu and X. Tan, “Metalloproteins/Metalloenzymes for the Synthesis of Acetyl-CoA in the Wood-Ljungdahl Pathway,” Science in China, Series B: Chemistry 52, no. 12 (2009): 2071–2082.

[79] J. Tian, W. Deng, Z. Zhang, et al., “Discovery and Remodeling of Vibrio natriegens as a Microbial Platform for Efficient Formic Acid Biorefinery,” Nature Communications 14, no. 1 (2023): 7758.

[80] X. Lu, Y. Liu, Y. Yang, et al., “Constructing a Synthetic Pathway for Acetylcoenzyme a From One-Carbon Through Enzyme Design,” Nature Communications 10 (2019): 1378.

[81] K. Schneider, R. Peyraud, P. Kiefer, et al., “The Ethylmalonyl-CoA Pathway is Used in Place of the Glyoxylate Cycle by Methylobacterium extorquens AM1 During Growth on Acetate,” Journal of Biological Chemistry 287, no. 1 (2012): 757–766.

[82] A. Bar-Even, E. Noor, A. Flamholz, and R. Milo, “Design and Analysis of Metabolic Pathways Supporting Formatotrophic Growth for Electricity-Dependent Cultivation of Microbes,” Biochimica et Biophysica Acta (BBA)—Bioenergetics 1827, no. 8–9 (2013): 1039–1047.

[83] J. Bertsch and V. Müller, “Co Metabolism in the Acetogen Acetobacterium woodii,” Applied and Environmental Microbiology 81, no. 17 (2015): 5949–5956.

[84] V. Müller, F. Imkamp, E. Biegel, S. Schmidt, and S. Dilling, “Incredible Anaerobes: From Physiology to Genomics to Fuels,” Annals of the New York Academy of Sciences 1125, no. 1 (2008): 1–373.

[85] S. Kim, N. Giraldo, V. Rainaldi, et al., “Optimizing E. coli as a Formatotrophic Platform for Bioproduction via the Reductive Glycine Pathway,” Frontiers in Bioengineering and Biotechnology 11 (2023): 1091899.

[86] Y. Hong, P. Arbter, W. Wang, L. N. Rojas, and A. P. Zeng, “Introduction of Glycine Synthase Enables Uptake of Exogenous Formate and Strongly Impacts the Metabolism in Clostridium pasteurianum,” Biotechnology and Bioengineering 118, no. 3 (2021): 1366–1380.

[87] H. Li, P. H. Opgenorth, D. G. Wernick, et al., “Integrated Electromicrobial Conversion of CO₂ to Higher Alcohols,” Science 335, no. 6076 (2012): 1596.

[88] Q. Yang, X. Guo, Y. Liu, and H. Jiang, “Biocatalytic C-C Bond Formation for One Carbon Resource Utilization,” International Journal of Molecular Sciences 22, no. 4 (2021): 1890.

[89] N. J. Claassens, G. Bordanaba-Florit, C. A. R. Cotton, et al., “Replacing the Calvin Cycle With the Reductive Glycine Pathway in Cupriavidus necator,” Metabolic Engineering 62 (2020): 30–41.

[90] D. Zhang, P. Clauwaert, A. Luther, et al., “Sub- and Supercritical Water Oxidation of Anaerobic Fermentation Sludge for Carbon and Nitrogen Recovery in a Regenerative Life Support System,” Waste Management 77 (2018): 268–275.

[91] A. Satanowski, B. Dronsella, E. Noor, et al., “Awakening a Latent Carbon Fixation Cycle in Escherichia coli,” Nature Communications 11, no. 1 (2020): 5812.

[92] T. Schwander, L. Schada von Borzyskowski, S. Burgener, N. S. Cortina, and T. J. Erb, “A Synthetic Pathway for the Fixation of Carbon Dioxide In Vitro,” Science 354, no. 6314 (2016): 900–904.

[93] J. Wang, K. Anderson, E. Yang, L. He, and M. E. Lidstrom, “Enzyme Engineering and In Vivo Testing of a Formate Reduction Pathway,” Synthetic Biology 6, no. 1 (2021): 1–14.

[94] L. Zelcbuch, S. N. Lindner, Y. Zegman, et al., “Pyruvate Formate-Lyase Enables Efficient Growth of Escherichia coli on Acetate and Formate,” Biochemistry 55, no. 17 (2016): 2423–2426.

[95] H. Yurimoto, N. Kato, and Y. Sakai, “Assimilation, Dissimilation, and Detoxification of Formaldehyde, a Central Metabolic Intermediate of Methylotrophic Metabolism,” The Chemical Record 5, no. 6 (2005): 367–375.

[96] J. Bang, J. H. Ahn, J. A. Lee, et al., “Synthetic Formatotrophs for One-Carbon Biorefinery,” Advanced Science 8, no. 12 (2021): 2100199.

[97] A. L. Smith, “Development of a Novel Carbon Fixation Pathway Enabled by Protein Engineering,” (2015), 1–164.

[98] H. He, R. Höper, M. Dodenhöft, P. Marlière, and A. Bar-Even, “An Optimized Methanol Assimilation Pathway Relying on Promiscuous Formaldehyde-Condensing Aldolases in E. coli,” Metabolic Engineering 60 (2020): 1–13.

[99] M. Jia, M. Liu, J. Li, et al., “Formaldehyde: An Essential Intermediate for C1 Metabolism and Bioconversion,” ACS Synthetic Biology 13, no. 11 (2024): 3507–3522.

[100] X. Zhang, M. Li, Y. Xu, J. Ren, and A. P. Zeng, “Quantitative Study of H Protein Lipoylation of the Glycine Cleavage System and a Strategy to Increase Its Activity by Co-Expression of LplA,” Journal of Biological Engineering 13 (2019): 32.

[101] C. H. Calvey, V. Sànchez i Nogué, A. M. White, et al., “Improving Growth of Cupriavidus Necator H16 on Formate Using Adaptive Laboratory Evolution-Informed Engineering,” Metabolic Engineering 75 (2023): 78–90.

[102] G. Hu, L. Guo, C. Gao, W. Song, L. Liu, and X. Chen, “Synergistic Metabolism of Glucose and Formate Increases the Yield of Short-Chain Organic Acids in Escherichia coli,” ACS Synthetic Biology 11, no. 1 (2022): 135–143.

[103] J. Bang, C. H. Hwang, J. H. Ahn, J. A. Lee, and S. Y. Lee, “Escherichia coli Is engineered to grow on CO₂ and formic acid,” Nature Microbiology 5, no. 12 (2020): 1459–1463.

[104] S. Alpdagtas and B. Binay, “NADP⁺-Dependent Formate Dehydrogenase: A Review,” Biocatalysis and Biotransformation 39, no. 4 (2021): 260–268.

[105] D. Jin, X. R. Bao, J. H. Yang, et al, “Integrated and Enhanced Coassimilation of Formate and Acetate for Efficient Utilization of Lignocellulosic Hydrolysate,” ACS Sustainable Chemistry & Engineering 12, no. 7 (2024): 2802–2812.

[106] L. Calzadiaz-Ramirez, C. Calvó-Tusell, G. M. M. Stoffel, et al., “In Vivo Selection for Formate Dehydrogenases With High Efficiency and Specificity Toward NADP,” ACS Catalysis 10, no. 14 (2020): 7512–7525+.

[107] R. Boonkumkrong, P. Chunthaboon, P. Munkajohnpong, et al., “A High Catalytic Efficiency and Chemotolerant Formate Dehydrogenase From Bacillus simplex,” Biotechnology Journal 19, no. 1 (2024): e2300330.

[108] A. Andreadeli, D. Platis, V. Tishkov, V. Popov, and N. E. Labrou, “Structure-Guided Alteration of Coenzyme Specificity of Formate Dehydrogenase by Saturation Mutagenesis to Enable Efficient Utilization of NADP⁺,” FEBS Journal 275, no. 15 (2008): 3859–3869.

[109] J. Zheng, T. Yang, J. Zhou, M. Xu, X. Zhang, and Z. Rao, “Elimination of a Free Cysteine by Creation of a Disulfide Bond Increases the Activity and Stability of Candida boidinii Formate Dehydrogenase,” Applied and Environmental Microbiology 83, no. 2 (2017): e02624-02616.

[110] S. K. Bhatia, R. K. Bhatia, J. M. Jeon, G. Kumar, and Y. H. Yang, “Carbon Dioxide Capture and Bioenergy Production Using Biological System—A Review,” Renewable & Sustainable Energy Reviews 110 (2019): 143–158.

[111] E. Sapountzaki, U. Rova, P. Christakopoulos, and I. Antonopoulou, “Renewable Hydrogen Production and Storage via Enzymatic Interconversion of CO₂ and Formate With Electrochemical Cofactor Regeneration,” Chemsuschem 16, no. 17 (2023): e202202312.

[112] A. K. Rai, T. Chen, and J. V. Moroney, “Mitochondrial Carbonic Anhydrases Are Needed for Optimal Photosynthesis at Low CO₂ Levels in Chlamydomonas,” Plant Physiology 187, no. 3 (2021): 1387–1398.

[113] J. Jiang, X. Li, K. Yang, et al., “Formate for Enhancing the Growth of Microalgae and Accumulating High-Value Products,” Algal Research 75 (2023): 103261.

[114] H. Zhang, Y. Li, J. Nie, J. Ren, and A. P. Zeng, “Structure-Based Dynamic Analysis of the Glycine Cleavage System Suggests Key Residues for Control of a Key Reaction Step,” Communications Biology 3, no. 1 (2020): 756.

[115] M. Dragosits and D. Mattanovich, “Adaptive Laboratory Evolution—Principles and Applications for Biotechnology,” Microbial Cell Factories 12 (2013): 64.

[116] M. Mavrommati, A. Daskalaki, S. Papanikolaou, and G. Aggelis, “Adaptive Laboratory Evolution Principles and Applications in Industrial Biotechnology,” Biotechnology Advances 54 (2022): 107795.

[117] M. Wang, H. Wang, C. Gao, et al., “Efficient Production of Protocatechuic Acid Using Systems Engineering of Escherichia coli,” Metabolic Engineering 82 (2024): 134–146.

[118] Z. Liu, T. Oyetunde, W. D. Hollinshead, et al., “Exploring Eukaryotic Formate Metabolisms to Enhance Microbial Growth and Lipid Accumulation,” Biotechnology for Biofuels 10 (2017): 22.

[119] H. Gevorgyan, A. Trchounian, and K. Trchounian, “Formate and Potassium Ions Affect Escherichia coli Proton ATPase Activity at Low pH During Mixed Carbon Fermentation,” IUBMB Life 72, no. 5 (2020): 915–921.

[120] N. J. Claassens, C. A. R. Cotton, D. Kopljar, and A. Bar-Even, “Making Quantitative Sense of Electromicrobial Production,” Nature Catalysis 2, no. 5 (2019): 437–447.

[121] M. Stöckl, N. Claassens, S. Lindner, E. Klemm, and D. Holtmann, “Coupling Electrochemical CO₂ Reduction to Microbial Product Generation—Identification of the Gaps and Opportunities,” Current Opinion in Biotechnology 74 (2022): 154–163.

[122] A. Garron and F. Epron, “Use of Formic Acid as Reducing Agent for Application in Catalytic Reduction of Nitrate in Water,” Water Research 39, no. 13 (2005): 3073–3081.

[123] P. Yue, Z. Kang, Q. Fu, et al., “Life Cycle and Economic Analysis of Chemicals Production via Electrolytic (Bi)Carbonate and Gaseous CO₂ Conversion,” Applied Energy 304 (2021): 117768.

[124] Y. Ahn, J. Byun, D. Kim, B. S. Kim, C. S. Lee, and J. Han, “System-Level Analysis and Life Cycle Assessment of CO₂ and Fossil-Based Formic Acid Strategies,” Green Chemistry 21, no. 12 (2019): 3442–3455.

[125] L. Semenec, A. E. Laloo, B. L. Schulz, I. A. Vergara, P. L. Bond, and A. E. Franks, “Deciphering the Electric Code of Geobacter sulfurreducens in Cocultures With Pseudomonas aeruginosa via Swath-Ms Proteomics,” Bioelectrochemistry 119 (2018): 150–160.

[126] G. Tomazetto, S. Hahnke, D. E. Koeck, et al., “Complete Genome Analysis of Clostridium bornimense Strain M2/40T: A New Acidogenic Clostridium Species Isolated From a Mesophilic Two-Phase Laboratory-Scale Biogas Reactor,” Journal of Biotechnology 232 (2016): 38–49.

[127] M. K. F. Mohr, A. Satanowski, S. N. Lindner, T. J. Erb, and J. N. Andexer, “Rewiring Escherichia coli to Transform Formate Into Methyl Groups,” Microbial Cell Factories 24, no. 1 (2025): 55.

[128] A. Tramonti, E. Cuyàs, J. Encinar, et al., “Metformin Is a Pyridoxal-5′-Phosphate (PLP)-Competitive Inhibitor of SHMT2,” Cancers 13, no. 16 (2021): 4009.

[129] B. Sabel-Becker, N. P. Jost, A. K. Kaster, and D. Holtmann, “A Co-Feeding Strategy of Formate and H₂ for Methanogens-Enhancing Growth Parameters and Methane Production,” Journal of CO2 Utilization 93 (2025): 103049.

[130] K. Trchounian, S. Mirzoyan, A. Poladyan, and A. Trchounian, “Hydrogen Production by Escherichia coli Growing in Different Nutrient Media With Glycerol: Effects of Formate, pH, Production Kinetics and Hydrogenases Involved,” International Journal of Hydrogen Energy 42, no. 38 (2017): 24026–24034.

[131] K. Wang, Z. Wu, J. Du, et al., “Metabolic Engineering of Saccharomyces cerevisiae for Conversion of Formate and Acetate into Free Fatty Acids,” Fermentation 9, no. 11 (2023): 984.

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