Biochar (BC) is a material that has many applications in agricultural and environmental activities. The aim of the study was to define the influence of BC produced in low-temperature pyrolysis from various organic waste materials, including one-month-old compost (OMOC), pine bark (PB), pine needle mulch (NM), pine cones (PC) and maple leaves (ML), on soil enzyme activity as well as its relation with organic matter properties. A 60-day incubation pot experiment was set up to investigate the influence of BC amendment on soil (S) characteristics. After incubation, we investigated the activity of soil enzymes, the content of available phosphorus (AP), potassium (AK) and magnesium (AMg), total organic carbon (TOC), total nitrogen (TN), dissolved organic matter (DOM) and its fractional composition (content and share of carbon and nitrogen of humic (CHAs, NHAs) and fulvic (CFAs, NFAs) acids and humin fractions). The effect of the amended biochars differed depending on the feedstock material. In general, the use of biochar enriched the soil with AP, AK and AMg increased the soil carbon stock, increased the intensity of nitrogen transformation and influenced the soil enzyme activity. OMOC and ML biochars significantly increased soil fertility, which was expressed by the high value of the CHA/CFA ratio.