Physical properties, chemical composition, minerals content, amino acids profile and anti-nutritional factors i.e. phytic acid, tannins and trypsin inhibitor of lima bean seeds (Phaseolus lunatus) which newly cultivated in Egypt were investigated. Addition to study the effect of common processing methods (soaking, cooking, roasting, and dehulling) on the chemical composition, minerals content and anti-nutritional factors of lima bean seeds in order to be used in biscuit preparation. Results showed that protein content was significantly (P ≤ 0.05) increased in dehulled Lima bean (27.06%) compared to raw lima bean (26.02%). All processing methods significantly (P ≤ 0.05) increased Soluble Dietary Fiber (SDF), however soaked seeds contained the highest significant Total Dietary Fiber (TDF) (30.18%) and Insoluble Dietary Fiber (IDF) (22.15%). Raw lima bean was superior in calcium, iron, magnesium, sodium, potassium, copper, and zinc compared with all processed samples. Raw lima bean had a higher content of all essential amino acids, except methionine compared to wheat flour of 72% extraction rate. The effect of different processing methods used in this study on lowering phytic acid and tannins, was arranged in the following significant (P ≤ 0.05) order: dehulling > cooking> roasting > soaking. Trypsin inhibitor was completely eliminated by cooking and dehulling treatments. Results showed that no significant differences (P ≤ 0.05) were found in appearance, color, texture and overall acceptability of substituted with 30% raw or processed lima bean and between control biscuit (100% wheat flour). The diameter, spread ratio, chemical composition (protein, ash, crude fiber, SDF, IDF and TDF) and minerals were significantly increased in all substituted biscuits compared to control biscuit. Improvement in the essential amino acid profile of raw Lima biscuit was observed with higher values of essential amino acids, Chemical Score (CS), and Biological Value (BV). This study recommended the use of lima How to cite this paper: El-Gohery, S.S. (2021) Effect of Different Treatments on Nutritional Value of Lima Bean (Phaseolus lunatus) and Its Utilization in Biscuit Manufacture. Food and Nutrition Sciences, 12, 372-391.