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.