Soil organic matter largely controls the ecosystem functions of soil and is a source of energy and a nutrient substrate for the soil microbial community as well. The postagrogenic dynamics of organic matter stock (content of organic carbon, Corg; total nitrogen, Ntot; and their ratio, C/N), basic soil properties, and indicators of soil microbial and enzyme activities are analyzed for the chronosequence of Retic Albic Podzol (southern taiga zone, Kostroma oblast, Russia). Different plots of the chronosequence have been withdrawn from agricultural use 11, 16, and 40 years ago. The stratification ratios (SRs) 0–5 : 5–10, 0–10 : 10–20, and 0–10 : 20–30 cm layers for Corg and Ntot contents are also estimated. A statistically significant increase in the contents of Corg and Ntot during the postagrogenic succession is observed in the topsoil layer of the old arable horizon (0–5 and 5–10 cm) as well as an increase in the stratification of their distribution, especially after a closed tree canopy has developed on the former arable land. Postagrogenic succession promotes an increase in the soil basal respiration rate and the content of microbial biomass, the maximum values of which are most often recordable in the soil under a 40-year-old secondary small-leaved forest. During the natural reforestation of the former arable lands, a statistically significant increase in the phosphatase activity is observed within the entire old arable horizon. The peaks of oxidoreductase activity correspond to the stages of succession with the maximum uptake of easily decomposable plant litter. Over the 40 years of postagrogenic succession, all analyzed parameters (except for pHKCl) have increased in a statistically significant manner at all considered depths of the old arable layer, with the highest annual increase in 0–5-cm layer, most enriched for organic matter.