Influence of chemical composition of sawdust on the nutritional profile of oyster mushrooms (Pleurotus ostreatus) has yet to receive significant research attention. This information will help mushroom growers to select specific sawdust for the production of mushroom with desired dietary preferences. This study assessed the influence of the chemical composition of sawdust on the macronutrients and ash content of the pearl oyster mushrooms. The American Standard for Testing Materials and other widely accepted protocols were used to determine the C-N ratio, pH, lignin, hemicellulose and cellulose contents of mixed sawdust from tropical wood species. The study evaluated the fat, crude fibre, crude protein, carbohydrate, and ash content of the oyster mushroom cultivated on the sawdust. Cellulose constituted the largest component of the sawdust (47.82%), followed by lignin (33.29%). The yield of the mushroom (on 0.05 kg of sawdust) ranged from 490.1 to 540.9 g (biological efficiency: 44–50%); the average carbohydrates constituent in the mushroom was 56.28%. pH of the sawdust influenced the crude protein, carbohydrate, fat and ash content of oyster mushrooms (p<0.05) most significantly. The hemicelluloses also had a significant effect (p<0.05) on the mushroom’s minerals, fat and crude fiber content. The study revealed that the mushroom producers would likely obtain high protein content using sawdust with low pH (slightly acidic to slightly basic) in the oyster mushroom. Mushrooms grown on substrates, rich in hemicelluloses, had low fat and high crude fiber content.