Unpretreated spent beer grains were successfully used as a basic substrate material for the cultivation of Pleurotus ostreatus. The effects of spent grain types, additives, substrate moisture content, and substrate packing density on the yield and nutrition of fruit bodies were investigated. The cultivation results showed that few fruit bodies were formed on spent grain alone; however, a significantly high biological efficiency (19.1%) was obtained with the addition of wheat bran to (45%). The chemical analysis of fruit bodies indicated that P. ostreatus cultivated on spent grain substrate had a higher nutritional value than those grown on other reported types of substrates. The total amino acid content in the fruit bodies was 347.5 mg/g dry matter, and the crude protein content was as high as 53.3% on a dry weight basis. It was also found that the cultivation of P. ostreatus increased the crude protein content, while it decreased the ratio of lignin to cellulose, of the spent grain substrate