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The vitamin content of cultivated mushrooms differs from one species to another, depending on their stage of development, the nutrient substrate used to produce them, and the microclimate in the culture space. Agaricus blazei Murrill is one of the most popular cultivated medicinal mushrooms, with scientifically proven therapeutic properties. Considering that the Agaricus spp. mushrooms culture substrate can be produced using various raw materials, in this paper we have studied the influence of the culture substrate using four types of substrate with different protein additions on the vitamin content of mushrooms. The food qualities of the Agaricus blazei Murrill mushrooms, evaluated by the chemical composition, generally revealed the product obtained on the classic compost, improved with the addition of proteinaceous of corn flour. Mushrooms harvested on this substrate have the highest levels of B 1 (1151 μg 100g -1 dm), B 9 (671 μg 100g -1 dm), B 12 (906 μg 100g -1 dm), PP (55.33 μg 100g -1 dm), and C vitamins (21.67 μg 100g -1 dm). The content of ergosterol, as a precursor of D 2 vitamin, has higher values ​​in the product obtained on the classic compost, with the addition of wheat bran (90.17 mg 100g -1 dm) and the addition of corn flour (94 mg 100g -1 dm)., Competing Interests: “The authors declare no conflict of interest.”