In daily life, energy plays a critical role. Hydrogen energy is widely recognized as one of the cleanest energy carriers available today. However, hydrogen must be produced as it does not exist freely in nature. Various methods are available for hydrogen production, including electrolysis, thermochemical technology, and biological methods. This study explores the production of biological hydrogen through the degradation of organic substrates by anaerobic microorganisms. Bacillus paramycoides and Cereibacter azotoformans strains were selected as they have not yet been studied for biological hydrogen fermentation. This study investigates the ability of these microorganisms to produce biological hydrogen. Initially, the cells were identified using cell morphology study, gram staining procedure, and 16S ribosomal RNA (rRNA) gene polymerase chain reaction. The cells were revealed as Bacillus paramycoides (MCCC 1A04098) and Cereibacter azotoformans (JCM 9340). Moreover, the growth behaviour and biological hydrogen production of the dark and photo fermentative cells were studied. The inoculum concentrations experimented with were 1% and 10% inoculum size. This study found that Bacillus paramycoides and Cereibacter azotoformans are promising strains for hydrogen production, but further optimization processes should be performed to obtain the highest hydrogen yield.

Industrial revolution has created high dependent on fossil fuels for energy creation. However, combustion of fossil fuels has created excessive amount of greenhouse gases, hence led to climate change. Thus, renewable energy has been proposed to alleviate the environmental pollution issues around the globe. One of the promising renewable energies is green hydrogen energy. Commercialized technologies such as electrolysis and thermochemical reaction are utilized to form hydrogen energy. Nonetheless, these processes require high energy and yet producing greenhouse gases that harm the environment. In this study, biodegradation process to produce hydrogen energy has been explored. To our knowledge, Bacillus paramycoides strain has not yet been investigated for biological hydrogen evolution. Therefore, in this paper, the ability of Bacillus paramycoides to produce biological hydrogen has been studied. The rod-shaped and gram-positive Bacillus paramycoides was identified under scanning electron microscope and gram staining procedure. Furthermore, biological hydrogen generation by Bacillus sp. was experimented for 96 hours. The result shows that 4668 ± 120 ppm cumulative hydrogen gas was generated through dark fermentation process. For Bacillus sp. growth study, lag, log, and stationary phase have been achieved in 96 hours. In a summary, metabolic engineering to degrade abundant biomass wastes is a sustainable pathway to produce hydrogen energy, simultaneously resolve waste management issue around the globe.