Agaricus bohusii Bon is an edible and prized mushroom especially common in Serbia and southern Europe. Herein, this species was chemically characterized by evaluation of nutritional value (e.g. macromolecules, free sugars and fatty acids), bioactive compounds (e.g. tocopherols, phenolic compounds and organic acids), and antioxidant activity of its methanolic extract (e.g. scavenging activity, reducing power and inhibition of lipid peroxidation). Its antifungal preserving properties were also evaluated after incorporation of A. bohusii extract in cream cheese, using fungus Penicillium verucosum var. cyclopium as source contaminant. Comparison of sensory evaluation of cream cheese alone and enriched with A. bohusii extract was recorded. According to our findings, A. bohusii was rich source of carbohydrates and proteins, containing γ-tocopherol as the only isoform of tocopherols. Polyunsaturated fatty acids also predominated over mono and unsaturated fatty acids. p-Hydroxybenzoic and p-coumaric acids were the phenolic acids identified in the studied sample; two related compounds were found in higher amounts: γ-L-glutaminyl-4-hydroxybenzene and cinnamic acid. Malic, oxalic and fumaric acids were the organic acids identified and quantified in A. bohusii. High concentration of total phenolics was in correlation with strong antioxidant capacity. Methanol extract successfully inhibited development of P. verucosum var. cyclopium in cream cheese, tested at room temperature after 7 days of inoculation. Sensory evaluation showed cream cheese in combination with A. bohusii extract slightly more acceptable to panelists than cream cheese alone.