Cholesterol oxidase is widely used in clinical diagnostics for measuring cholesterol levels in blood samples. It plays a crucial role in enzymatic assays to determine total cholesterol, aiding in the assessment of cardiovascular health.

In the food industry, this enzyme is utilized to detect cholesterol content in dairy products and other food items, ensuring quality control and safety.

Additionally, cholesterol oxidase has applications in biotechnology research, particularly in studying lipid metabolism and developing biosensors for cholesterol detection.

Its ability to oxidize cholesterol also makes it valuable in producing steroid intermediates for pharmaceutical manufacturing.

In agriculture, cholesterol oxidase is employed in the development of biopesticides to control populations of harmful insects by disrupting their lipid metabolism.