The antimicrobial peptides derived from microalgae have attracted a huge attention due to the insufficient availability of effective drugs from the natural resources. In this study, the enzymatic hydrolysate of protein derived from Limnospira maxima was prepared using pepsin under optimized conditions. The peptides with range of 10kDa were isolated and purified using the Ultra membrane filtration, SDS-PAGE, and TLC. Furthermore, the peptide sequence was identified and characterized by MALDI-TOF mass spectrometry in which an algal peptide, KLENCNYAVELGK showed a strong signal at 466.68m/z among seven peptides derived from the pepsin hydrolysate. The FT-IR spectroscopic study confirmed the presence of a characteristic functional group of amino acids in the sequence. The algal derived peptide showed antibacterial properties against Escherichia coli (27 ± 0.5mm) and Staphylococcus aureus (14mm ± 0.5). This study paves a way to explore the antibacterial peptide from a novel species, L. maxima (MZ26519) evident to utilize for the novel drug to overcome the conventional approach.