The results of studies on the effect of the substrate composition for the cultivation of Lion’s mane mushroom (Hericium erinaceus) are presented. The search for alternative raw materials for the preparation of substrates is associated with the possibility of using lignin-ammonium compost, balanced in terms of basic nutrients, which is much cheaper than oak sawdust and is more accessible in the region. Replacing brown rice by wheat grain in the substrate contributed to the enrichment of the mass with vitamins, proteins and essential amino acids. The possibility of using a lignin component in the substrate composition in the form of ligninammonia compost (50%) and wheat grain (20%), partially replacing expensive and scarce raw materials (oak sawdust, brown rice), was evaluated. The use of a new substrate composition promotes the active colonization of the substrate mass by the mycelium of medicinal Lion’s mane mushroom of the Beard and Belgium strains (97–99%), additional (by 0.39% of the substrate mass) collection of fruiting bodies of the Beard strain and increased (by 1.6%) yield of dry mass of products of the Beard and Belgium strains. The economic effect of growing H. erinaceus on the developed substrate is expressed in profit (for the Beard strain – 20,540 rubles, for the Belgium strain – 5,860 rubles), while on the classical substrate a loss is obtained. The profitability of the mushroom production of the studied strains on the experimental substrate is 16.4% for the Beard strain and 4.68% for the Belgium strain, on the classical substrate – 12.1% and 21.0%, respectively. Reducing the cost of mushroom products may have a positive effect on the purchasing power of consumers. The addition of a lignin component to the substrate promotes the utilization of of hydrolytic lignins in large quantities, thus improving the environmental situation in the places where they are stored.