Clotridial hydrogenases and the biohydrogen production

Among the large variety of microorganisms capable of fermentative hydrogen production, strict anaerobes such as Clostridium spp. are one of the most widely studied. They produce hydrogen by butyric and mixed-acid fermentations at optimal pH values ranging from 4.5 to 5.5. While fermentative conditions such as substrate type, pH, hydraulic and solid retention time, H2 partial pressure and the concentration of acids produced have been extensively studied and optimized, relatively little is known about the different forms of hydrogenases present in clostridia. Building on previous reports [1, 2] and by analyzing sequenced genomes, we found that [FeFe] hydrogenases are not a homogenous group of enzymes, but exist in multiple forms with different modular structures and are especially abundant in Clostridum spp. [3].
However, among the numerous studies performed on fermentative hydrogen production by Clostridium sp., only a few are specifically concerned with hydrogenases. Even there the authors focus on one type of [FeFe] hydrogenase, (CpI-like) without considering the existence of multiple forms of this enzyme within one species. Therefore, we focused our research on the better characterization of different forms of hydrogenases present in the genus Clostridium. Using newly designed degenerate primers, specific for clostridial hydrogenases, we amplified different hydrogenases from our species of interest. Further, by designing specific qPCR assays we have quantitatively targeted different hydrogenases. By analyzing differential gene expression, according to applied growth conditions, we believe to optimize the hydrogen production process in order to achieve better production rates.
To conclude, we think that a a precise knowledge of hydrogen metabolism and hydrogenases is essential to optimization of the biohydrogen production process and should therefore be a goal for future research.