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Hydrogen Production by Immobilized Cells of Clostridium intestinale Strain URNW Using Alginate Beads

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Abstract

Biological hydrogen (H2) is a promising candidate for production of renewable hydrogen. Using entrapped cells rather than conventional suspended cell cultures for the production of H2 offers several advantages, such as improved production yields related to higher cell density, and enhanced resistance to substrate and end-product inhibition. In this study, H2 production by a novel isolate of Clostridium intestinale (strain URNW) was evaluated using cells entrapped within 2% calcium-alginate beads under strictly anaerobic conditions. Both immobilized cells and suspended cultures were studied in sequential batch-mode anaerobic fermentation over 192 h. The production of H2 in the headspace was examined for four different initial cellobiose concentrations (5, 10, 20, and 40 mM). Although a lag period for initiation of the fermentation process was observed for bacteria entrapped within hydrogel beads, the immobilized cells achieved both higher volumetric production rates (mmol H2/(L culture h)) and molar yields (mol H2/mol glucose equivalent) of H2 compared with suspended cultures. In the current study, the maximum cellobiose consumption rate of 0.40 mM/h, corresponding to 133.3 mg/(L h), was achieved after 72 h of fermentation by immobilized cells, generating a high hydrogen yield of 3.57 mol H2/mol cellobiose, whereas suspended cultures only yielded 1.77 mol H2/mol cellobiose. The results suggest that cells remain viable within the hydrogels and proliferated with a slow rate over the course of fermentation. The stable productivity of immobilized cells over 8 days with four changes of medium depicted that the immobilized cells of the isolated strain can successfully yield higher hydrogen and lower soluble metabolites than suspended cells suggesting a feasible process for future applications for bioH2 production.

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Acknowledgements

AT gratefully acknowledges the financial support of NSERC through PDF program.

Funding

This work was funded by The Scientific And Technological Research Council Of Turkey (TUBITAK, BIDEB 2214), and Natural Sciences and Engineering Research Council of Canada (NSERC), through the Strategic Programs grant (STPGP 306944-04), by Genome Canada, through the Applied Genomics Research in Bioproducts or Crops (ABC) program for the grant titled, “Microbial Genomics for Biofuels and Co-Products from Biorefining Processes.”

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Authors and Affiliations

  1. Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba, Canada

    Mine Güngörmüşler, Ali Tamayol & David B. Levin

  2. Department of Genetics and Bioengineering, Izmir University of Economics, Balçova, Izmir, Turkey

    Mine Güngörmüşler

  3. Department of Biomedical Engineering, University of Connecticut Health Center, Farmington, CT, USA

    Ali Tamayol

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  1. Mine Güngörmüşler
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  3. David B. Levin
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Contributions

MG and AT designed and investigated the study. MG validated the experiments. MG prepared the original draft. DBL administrated the project and provided funding. AT and DBL reviewed and edited the manuscript. MG, AT, and DBL read the manuscript. MG, AT, and DBL approved the submission of the manuscript to the journal.

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Correspondence to Mine Güngörmüşler.

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Highlights

• C. intestinale URNW entrapped in sodium alginate successfully produced biohydrogen.

• Higher molar yields in mol H2/mol hexose eq. were achieved with immobilized cells.

• The immobilized cells showed stability during sequential batch-mode over 192 h.

• Four different initial cellobiose concentrations were tested for both cultures.

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Güngörmüşler, M., Tamayol, A. & Levin, D.B. Hydrogen Production by Immobilized Cells of Clostridium intestinale Strain URNW Using Alginate Beads. Appl Biochem Biotechnol 193, 1558–1573 (2021). https://doi.org/10.1007/s12010-021-03503-1

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