Chitin deacetylase is an enzyme produced by several organisms, such as bacteria, fungi, and insects. In most cases, the enzyme is produced intracellularly; however, some fungi also produce the enzyme extracellularly, so the work aimed to compare the activity and the energies of activation of intracellular and extracellular chitin deacetylase. The enzymes produced by Absidia orchidis vel coerulea were used in experiments. It was shown that intracellular chitin deacetylase is significantly more active than the extracellular form. The activation energies were evaluated as 77.77 kJ/mol and 90.88 kJ/mol for intracellular and extracellular chitin deacetylase, respectively, meaning that intracellular enzymes act faster. Our paper is the first one that presents the energy of activation of chitin deacetylase. The presented energy of activation evaluation and comparison method can be used as a good tool to choose the chitin deacetylase for industrial application when the enzyme is available in different forms or comes from different sources.