Bioscience Evidence 2024, Vol.14, No.2, 81-92 http://bioscipublisher.com/index.php/be 91 8 Concluding Remarks The exploration of metabolic pathways and genetic engineering in anaerobic bacteria for biohydrogen production has revealed significant advancements and highlighted ongoing challenges. As the global demand for renewable energy sources continues to increase, biohydrogen production has emerged as a promising avenue for sustainable energy generation. This review has provided a comprehensive analysis of the metabolic pathways involved in biohydrogen production, including acidogenic and butyrate-type fermentations, and the critical role of hydrogenases in these processes. Genetic engineering techniques, such as CRISPR-Cas9, gene knockout, and synthetic biology approaches, have been instrumental in enhancing hydrogen yields by optimizing these pathways. The integration of omics technologies has provided deeper insights into the regulatory networks governing biohydrogen production and has facilitated the identification of new targets for genetic engineering. Additionally, the development of co-culture systems and microbial consortia, which leverage synergistic interactions between different microbial species, has further enhanced hydrogen production from complex substrates. Despite these advances, challenges such as the stability of engineered traits, economic scalability, and biosafety concerns remain significant hurdles to the widespread adoption of biohydrogen technology. Genetic engineering has undeniably played a pivotal role in advancing biohydrogen production, enabling the optimization of metabolic pathways and the creation of engineered strains with enhanced capabilities. The precise modification of hydrogen-producing bacterial genomes has opened new possibilities for improving efficiency, yield, and process stability. As the field progresses, the integration of synthetic biology and omics technologies will further expand the potential of genetic engineering. The design of synthetic metabolic pathways, the development of dynamic regulatory circuits, and the use of directed evolution to refine enzyme function are all promising areas of innovation. However, the successful commercialization of biohydrogen technology will require a holistic approach that addresses not only the technical challenges but also the economic and environmental aspects. Ensuring the biosafety of genetically modified organisms and minimizing the environmental impact of biohydrogen production are critical considerations that must be prioritized. In conclusion, the future of biohydrogen production is bright, with genetic engineering at the forefront of this exciting field. By continuing to innovate and address the challenges, biohydrogen has the potential to become a key player in the global transition to sustainable energy. Acknowledgments The author expresses gratitude to the two anonymous peer reviewers for their feedback. Conflict of Interest Disclosure The author affirms that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Amin M., Ghasemian M., Bina B., Taheri E., and Fatehizadeh A., 2019, Biohydrogen production as a clean fuel by acid and alkaline pretreated mixed culture during glucose fermentation, Health Scope, 8(2): e12903. https://doi.org/10.5812/JHEALTHSCOPE.12903 Aslam M., Ahmad R., Yasin M., Khan A., Shahid M., Hossain S., Khan Z., Jamil F., Rafiq S., Bilad M., Kim J., and Kumar G., 2018, Anaerobic membrane bioreactors for biohydrogen production: Recent developments, challenges and perspectives, Bioresource Technology, 269: 452-464. https://doi.org/10.1016/j.biortech.2018.08.050 Ben Gaida L., Gannoun H., Casalot L., Davidson S., and Liebgott P., 2022, Biohydrogen production by Thermotoga maritima from a simplified medium exclusively composed of onion and natural seawater, Comptes Rendus. Chimie., 25(S2): 129-143. https://doi.org/10.5802/crchim.136 Bengelsdorf F., Beck M., Erz C., Hoffmeister S., Karl M., Riegler P., Wirth S., Poehlein A., Weuster-Botz D., and Dürre P., 2018, Bacterial anaerobic synthesis gas (Syngas) and CO2+H2 fermentation, Advances in Applied Microbiology, 103: 143-221. https://doi.org/10.1016/bs.aambs.2018.01.002 Buckel W., 2021, Energy conservation in fermentations of anaerobic bacteria, Frontiers in Microbiology, 12: 703525. https://doi.org/10.3389/fmicb.2021.703525
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