Hydrogen metabolic patterns driven by Clostridium-Streptococcus community shifts in a continuous stirred tank reactor
The hydrogen (H2) production efficiency in dark fermentation systems is strongly dependent on the occurrence of metabolic pathways derived from the selection of microbial species that either consume molecular H2 or outcompete hydrogenogenic bacteria for the organic substrate. In this study, the effect of organic loading rate (OLR) on the H2 production performance, the metabolic pathways, and the microbial community composition in a continuous system was evaluated. Two bacterial genera, Clostridium and Streptococcus, were dominant in the microbial community depending on the OLR applied. At low OLR (14.7–44.1 gLactose/L-d), Clostridium sp. was dominant and directed the system towards the acetate-butyrate fermentation pathway, with a maximum H2 yield of 2.14 molH2/molHexose obtained at 29.4 gLactose/L-d. Under such conditions, the volumetric hydrogen production rate (VHPR) was between 3.2 and 11.6 LH2/L-d. In contrast, relatively high OLR (58.8 and 88.2 gLactose/L-d) favored the dominance of Streptococcus sp. as co-dominant microorganism leading to lactate production. Under these conditions, the formate production was also stimulated serving as a strategy to dispose the surplus of reduced molecules (e.g., NADH2+), which theoretically consumed up to 5.72 LH2/L-d. In such scenario, the VHPR was enhanced (13.7–14.5 LH2/L-d) but the H2 yield dropped to a minimum of 0.74 molH2/molHexose at OLR = 58.8 gLactose/L-d. Overall, this research brings clear evidence of the intrinsic occurrence of metabolic pathways detrimental for biohydrogen production, i.e., lactic acid fermentation and formate production, suggesting the use of low OLR as a strategy to control them.
KeywordsBiohydrogen Dark fermentation Lactic acid bacteria (LAB) Hydrogen-producing bacteria (HPB) Microbial community
Rodolfo Palomo-Briones is thankful for the PhD scholarship provided by the CONACYT. The authors acknowledge the technical assistance of Gaëlle Santa-Catalina, Dulce Partida Gutiérrez, Guillermo Vidriales Escobar, and Juan Pablo Rodas Ortiz. The infrastructure provided by the CONACYT INFR-2014-01-224220 is also acknowledged.
This research was financially supported by the Fondo de Sustentabilidad Energética SENER-CONACYT, Clúster Biocombustibles Gaseosos (project 247006).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The authors confirm that the article does not contain any studies with human participants or animals.
- Agler MT, Wrenn BA, Zinder SH, Angenent LT (2011) Waste to bioproduct conversion with undefined mixed cultures: the carboxylate platform. Trends Biotechnol 29:70–78. https://doi.org/10.1016/j.tibtech.2010.11.006
- APHA/AWWA/WEF (2012) Standard methods for the examination of water and wastewaterGoogle Scholar
- Azwar MY, Hussain MA, Abdul-Wahab AK (2014) Development of biohydrogen production by photobiological, fermentation and electrochemical processes: a review. Renew Sust Energ Rev 31:158–173. https://doi.org/10.1016/j.rser.2013.11.022
- Cabrol L, Marone A, Tapia-Venegas E, Steyer J-P, Ruiz-Filippi G, Trably E (2017) Microbial ecology of fermentative hydrogen producing bioprocesses: useful insights for driving the ecosystem function. FEMS Microbiol Rev 41(2):158–181. https://doi.org/10.1093/femsre/fuw043 CrossRefPubMedGoogle Scholar
- Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7(5):335–336. https://doi.org/10.1038/nmeth.f.303 CrossRefPubMedPubMedCentralGoogle Scholar
- Davila-Vazquez G, Alatriste-Mondragón F, de León-Rodríguez A, Razo-Flores E (2008) Fermentative hydrogen production in batch experiments using lactose, cheese whey and glucose: influence of initial substrate concentration and pH. Int J Hydrog Energy 33(19):4989–4997. https://doi.org/10.1016/j.ijhydene.2008.06.065 CrossRefGoogle Scholar
- Etchebehere C, Castelló E, Wenzel J, Anzola-Rojas, MP, Borzacconi L, Buitrón G, Cabrol L, Carminato VM, Carrillo-Reyes J, Cisneros-Pérez C, Fuentes L, Moreno-Andrade I, Razo-Flores E, Filippi GR, Tapia-Venegas E, Toledo-Alarcón J, Zaiat M (2016) Microbial communities from 20 different hydrogen-producing reactors studied by 454 pyrosequencing. Appl Microbiol Biotechnol 100(7):3371–3384. https://doi.org/10.1007/s00253-016-7325-y
- Gomes SD, Fuess LT, Mañunga T, Feitosa de Lima Gomes PC, Zaiat M (2016) Bacteriocins of lactic acid bacteria as a hindering factor for biohydrogen production from cassava flour wastewater in a continuous multiple tube reactor. Int J Hydrog Energy 41(19):8120–8131. https://doi.org/10.1016/j.ijhydene.2015.11.186 CrossRefGoogle Scholar
- Hung C-H, Cheng C-H, Guan D-W, Wang S-T, Hsu S-C, Liang C-M (2011) Interactions between Clostridium sp. and other facultative anaerobes in a self-formed granular sludge hydrogen-producing bioreactor. Int J Hydrog Energy 36(14):8704–8711. https://doi.org/10.1016/j.ijhydene.2010.06.010 CrossRefGoogle Scholar
- Köpke M, Held C, Hujer S, Liesegang H, Wiezer A, Wollherr A, Ehrenreich A, Liebl W, Gottschalk G, Dürre P (2010) Clostridium ljungdahlii represents a microbial production platform based on syngas. Proc Natl Acad Sci 107(29):13087–13092. https://doi.org/10.1073/pnas.1004716107 CrossRefPubMedPubMedCentralGoogle Scholar
- Kleerebezem R, Van Loosdrecht MCM (2010) A generalized method for thermodynamic state analysis of environmental systems. Crit Rev Environ Sci Technol 40(1):1–54Google Scholar
- Milne CB, Eddy JA, Raju R, Ardekani S, Kim P-J, Senger RS, Jin Y-S, Blaschek HP, Price ND (2011) Metabolic network reconstruction and genome-scale model of butanol-producing strain Clostridium beijerinckii NCIMB 8052. BMC Syst Biol 5:130. https://doi.org/10.1186/1752-0509-5-130, 1
- Napoli F, Olivieri G, Russo ME, Marzocchella A, Salatino P (2011) Continuous lactose fermentation by Clostridium acetobutylicum—assessment of acidogenesis kinetics. Bioresour Technol 102(2):1608–1614. https://doi.org/10.1016/j.biortech.2010.09.004
- Nasr N, Velayutham P, Elbeshbishy E, Nakhla G, El Naggar MH, Khafipour E, Derakhshani H, Levin DB, Hafez H (2015) Effect of headspace carbon dioxide sequestration on microbial biohydrogen communities. Int J Hydrog Energy 40(32):9966–9976. https://doi.org/10.1016/j.ijhydene.2015.06.077 CrossRefGoogle Scholar
- Noar J, Makwana ST, Bruno-Bárcena JM (2014) Complete genome sequence of solvent-tolerant Clostridium beijerinckii strain SA-1. Genome Announc. doi 2(6):e01310–e01314. https://doi.org/10.1128/genomeA.01310-14
- Nölling J, Breton G, Omelchenko MV, Kira S, Zeng Q, Gibson R, Lee HM, Dubois J, Qiu D, Hitti J, Sequencing GTC, Wolf YI, Tatusov RL, Sabathe F, Soucaille P, Daly MJ, Bennett GN, Koonin EV, Smith DR (2001) Genome sequence and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum. J Bacteriol 183:a823–4838. https://doi.org/10.1128/JB.183.16.4823 CrossRefGoogle Scholar
- R Development Core Team R (2011) R: a language and environment for statistical computingGoogle Scholar
- Shanmugam SR, Chaganti SR, Lalman JA, Heath DD (2014) Statistical optimization of conditions for minimum H2 consumption in mixed anaerobic cultures: effect on homoacetogenesis and methanogenesis. Int J Hydrog Energy 39(28):15433–15445. https://doi.org/10.1016/j.ijhydene.2014.07.143 CrossRefGoogle Scholar
- Si B, Li J, Li B, Zhu Z, Shen R, Zhang Y, Liu Z (2015) The role of hydraulic retention time on controlling methanogenesis and homoacetogenesis in biohydrogen production using upflow anaerobic sludge blanket (UASB) reactor and packed bed reactor (PBR). Int J Hydrog Energy 40(35):11414–11421. https://doi.org/10.1016/j.ijhydene.2015.04.035 CrossRefGoogle Scholar
- Sikora A, Baszczyk M, Jurkowski M, Zielenkiewicz U (2013) Lactic acid bacteria in hydrogen-producing consortia: on purpose or by coincidence? Lactic Acid Bacteria - R & D for Food, Health and Livestock Purposes. InTech, InGoogle Scholar