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Knowing the enemy: homoacetogens in hydrogen production reactors

  • Bioenergy and Biofuels
  • Published:
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Abstract

One of the bottlenecks of the hydrogen production by dark fermentation is the low yields obtained because of the homoacetogenesis persistence, a metabolic pathway where H2 and CO2 are consumed to produce acetate. The central reactions of H2 production and homoacetogenesis are catalyzed by enzyme hydrogenase and the formyltetrahydrofolate synthetase, respectively. In this work, genes encoding for the formyltetrahydrofolate synthetase (fthfs) and hydrogenase (hydA) were used to investigate the diversity of homoacetogens as well as their phylogenetic relationships through quantitative PCR (qPCR) and next-generation amplicon sequencing. A total of 70 samples from 19 different H2-producing bioreactors with different configurations and operating conditions were analyzed. Quantification through qPCR showed that the abundance of fthfs and hydA was strongly associated with the type of substrate, organic loading rate, and H2 production performance. In particular, fthfs sequencing revealed that homoacetogens diversity was low with one or two dominant homoacetogens in each sample. Clostridium carboxivorans was detected in the reactors fed with agave hydrolisates; Acetobacterium woodii dominated in systems fed with glucose; Blautia coccoides and unclassified Sporoanaerobacter species were present in reactors fed with cheese whey; finally, Eubacterium limosum and Selenomonas sp. were co-dominant in reactors fed with glycerol. Altogether, quantification and sequencing analysis revealed that the occurrence of homoacetogenesis could take place due to (1) metabolic changes of H2-producing bacteria towards homoacetogenesis or (2) the displacement of H2-producing bacteria by homoacetogens. Overall, it was demonstrated that the fthfs gene was a suitable marker to investigate homoacetogens in H2-producing reactors.

Key points

• qPCR and sequencing analysis revealed two homoacetogenesis phenomena.

• fthfs gene was a suitable marker to investigate homoacetogens in H2 reactors.

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Data availability

Sequence data that support the findings of this study have been deposited in GenBank with the following accession code: PRJNA659251.

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Funding

LF was funded by ANII and CAP-CSIC doctoral scholarship, and CE is funded by ANII-SNI. This work was partly supported by the Fondo Sectorial SENER-CONACYT Sustentabilidad Energética, Clúster Biocombustibles Gaseosos (project 247006). This work was also partly supported by CONACYT project A1-S-37174. The experimental work in Chile was financed by FONDECYT project 3160219 and FONDECYT project 11200211.

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Authors and Affiliations

  1. Laboratorio de Ecología Microbiana, Departamento de Bioquímica Y Genómica Microbiana, Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia, 3318, Montevideo, Uruguay

    Laura Fuentes & Claudia Ecthebehere

  2. División de Ciencias Ambientales, Instituto Potosino de Investigación Científica Y Tecnológica A.C, Camino a La Presa San José No, 2055, Col. Lomas 4a Sección, C.P., 78216, San Luis Potosí, SLP, México

    Rodolfo Palomo-Briones, José de Jesús Montoya-Rosales & Elías Razo-Flores

  3. Laboratorio Bioprocesos Ambientales, Facultad de Ingeniería, Instituto de Ingeniería Química, Universidad de La República, Herrera Y Reissig, 565, Montevideo, Uruguay

    Lucía Braga & Elena Castelló

  4. Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso, 2085, Valparaíso, Av. Brasil, Chile

    Alejandra Vesga

  5. Departamento de Medio Ambiente, Facultad de Ingeniería, Universidad de Playa Ancha Av, Leopoldo Carvallo 270, Valparaíso, Chile

    Estela Tapia-Venegas

Authors
  1. Laura Fuentes
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  2. Rodolfo Palomo-Briones
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  3. José de Jesús Montoya-Rosales
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  4. Lucía Braga
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  5. Elena Castelló
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  6. Alejandra Vesga
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  7. Estela Tapia-Venegas
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  8. Elías Razo-Flores
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  9. Claudia Ecthebehere
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Contributions

LF: conceptualization, data curation, formal analysis, methodology, writing—original draft, writing—review and editing. RP and JM: statistical analysis, data curation, formal analysis, writing—original draft, writing—review and editing. LB: designed and carried out some fermentation experiments and some molecular biology analysis. EC: formal analysis, data curation. AVB and ETV: designed and carried out some fermentation experiments and some molecular biology analysis and helped to review the manuscript. ERF: writing—review and editing, funding acquisition. CE: conceptualization, writing—review and editing.

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Correspondence to Claudia Ecthebehere.

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Fuentes, L., Palomo-Briones, R., de Jesús Montoya-Rosales, J. et al. Knowing the enemy: homoacetogens in hydrogen production reactors. Appl Microbiol Biotechnol 105, 8989–9002 (2021). https://doi.org/10.1007/s00253-021-11656-6

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