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Development and application of a functional CE-SSCP fingerprinting method based on [Fe–Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures

Abstract : A Capillary Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) method based on functional [Fe Fe]-hydrogenase genes was developed for monitoring the hydrogen (H-2)-producing clostridial population in mixed-culture bioprocesses. New non-degenerated primers were designed and then validated on their specific PCR detection of a broad range of clostridial hydA genes. The hydA-based CE-SSCP method gave a specific and discriminating profile for each of the Clostridium strains tested. This method was validated using H-2-producing mixed cultures incubated at temperatures ranging from 25 C to 45 degrees C. The hydA CE-SSCP profiles clearly differed between temperatures tested. Hence, they varied according to variations of the H-2 production performances. The HydA sequences amplified with the new primer set indicated that diverse Clostridium strains impacted the H-2 production yields. The highest performances were related to the dominance of Clostridium sporogenes-like hydA sequences. This CE-SSCP tool offers highly reliable and throughput analysis of the functional diversity and structure of the hydA genes for better understanding of the H-2-producing clostridial population dynamics in H-2 dark fermentation bioreactors.
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https://hal-amu.archives-ouvertes.fr/hal-01809619
Contributor : Marianne Quemeneur <>
Submitted on : Wednesday, June 6, 2018 - 9:35:55 PM
Last modification on : Saturday, October 3, 2020 - 3:22:06 AM

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Marianne Quéméneur, Jérôme Hamelin, Eric Latrille, Jean-Philippe Steyer, Eric Trably. Development and application of a functional CE-SSCP fingerprinting method based on [Fe–Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures. International Journal of Hydrogen Energy, Elsevier, 2010, 35 (24), pp.13158 - 13167. ⟨10.1016/j.ijhydene.2010.07.076⟩. ⟨hal-01809619⟩

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