Soil fertility is largely determined by the intensity and direction of enzymatic reactions. All biological processes associated with the transformation of substances and energy in the soil are carried out with the help of enzymes that play an important role in the mobilization of plant nutrients, and also determine the intensity and direction of the most important biochemical processes associated with the synthesis and decomposition of humus, hydrolysis of organic compounds and oxidative - the restorative regime of the soil, therefore, the enzymatic activity of the soil can be used as a diagnostic indicator of soil fertility under various types of anthropogenic load. The influence of the methods of basic soil cultivation on the enzymatic activity of ordinary chernozem in winter wheat crops during its cultivation on a pure and green manure fallow in the conditions of the Middle Volga region has been studied. According to the results of the studies, the highest catalytic activity of ordinary chernozem in winter wheat crops for the studied enzymes (urease, invertase, catalase, peroxidase and polyphenol oxidase) is confined to the 0-20 cm layer and is explained by the abundant location in it of the “living” root mass enriching the upper soil layer biologically active exudates that stimulate the development of microflora and activate enzymatic activity. Favorable conditions for the humification of plant residues and the accumulation of humus-like substances are formed when green manure fallow is included in the crop rotation and used as the main soil cultivation without loosening by 10-12 cm.