Protease is a well-known plant enzyme, used widely in the field of medicine and industry in various potential and economical applications. Available literatures show a lack of study related to the isolation of protease enzyme from Vicia faba L (Faba bean), hence, this study focuses on the isolation of protease from faba bean. Initially, the extraction process was carried out using overnight ammonium sulphate precipitation method, followed by protease assay to determine the enzyme activity in seed, leaf and pod samples of faba bean. Apparently, dialysis was carried out for partial purification, later High-Performance Liquid Chromatography (HPLC) for further purification. Finally, the molecular weight of the protease enzyme was determined from each sample extracted by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) respectively. The outcome of the study demonstrates that all the samples of faba bean contain protease enzyme with 60-70% saturation. The enzyme extracted from different parts of the sample show an optimum activity at pH 6 along with a molecular weight ranging between 60 to 100 kDa. In conclusion, the study suggests faba bean can serve as best source of protease enzyme. © 2018 Universiti Putra Malaysia.