Development and application of a functional CE-SSCP fingerprinting method based on [Fe–Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures - Aix-Marseille Université Access content directly
Journal Articles International Journal of Hydrogen Energy Year : 2010

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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hal-01809619 , version 1 (08-08-2023)

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Attribution - NonCommercial - NoDerivatives

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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, 2010, 35 (24), pp.13158 - 13167. ⟨10.1016/j.ijhydene.2010.07.076⟩. ⟨hal-01809619⟩
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