Nanoparticles represent a unique tool of manipulation of the biosynthetic activity of microorganisms, whose efficiency was confirmed on the biotechnological objects from different taxonomic groups, both abroad and in our country within the IMB of ASM. The correlations between the type, structure and sizes of nanoparticles and the level of biosynthesis/activity of fungal hydrolases present intrest, in order to decipher the mechanisms of influence of nanoparticles on biological objects and to elaborate new improved methods for obtaining of hydrolytic enzymes with application of nanoparticles as factor of influence. The research that was focused on assessing of the effect of nanoparticles with different composition and concentration on the micromycete Trichoderna koningii – producer of acid, neutral and alkaline proteases, revealed that ZnO nano-oxide can be used as potential stimulator of the enzymatic activity. High protease activity was observed for ZnO nanoparticles with dimensions of 30 nm and 60–70 nm that were added to the medium in the concentration of 5, 10 and 15 mg/L. The activity of neutral protease (the basic component of the enzyme complex of the strain), under the indicated conditions, has exceeded the control sample of about 2.5 times (with 255.0–258.3%), against a background of moderate increase in the activity of acid protease (with 16.6%) and alkaline protease (with 11.3%) (Figure). ZnO nanoparticles did not change the duration of the cultivation of micromycete Trichoderma koningii CNMN FD 15. The maximal biosynthesis of all three types of proteases was observed on the 9th day of cultivation and coincided with the maximal biosynthesis in the control sample.