The present study aimed to evaluate in vitro enzymatic activity of a novel biotechnological active complex based on natural origin compounds – thermophilic lipase and β-cyclodextrin – for hydrolysis of acylglycerides in plant oils and fats. β-cyclodextrin (β-CD) as an additive has attracted attention for enhance stability and efficiency of enzymes. In present study, the effects of β-CD on enzymatic hydrolysis of acylglycerides by thermophilic lipase were investigated by modern methods. The UV-spectroscopy, electron microscopy with TEM and kinetics of enzymatic hydrolysis were compared by the addition of β-CD, respectively. The results showed that lipase could produce the highest yield of oleic acid in presence of β-CD after 1 and 3h. The UV spectroscopy demonstrated that the absorbance and fluorescence of lipase decreased with increasing concentration of β-CD due to surface interaction and change of enzyme configuration. Moreover, electron microscopy with TEM showed that lipase formed a special active conglomerate with β-CD for improving hydrolysis and stability. Dermatology evaluation indicated that complex added to household products didn’t affect sensitive skin of hands. Overall results indicate that β-CD could increase enzymatic activity of the lipase against acylglycerides and can be considered as promising composition in ecological household products for regular hand application.