Abstract: In the last years, microbial caffeinases have been the subject of several studies due to their potential commercial importance and complexity as catalytic molecules. Caffeinases are capable of hydrolyzing caffeine, which represent the main chemical group of natural alkaloids occurring in the plants, particularly in coffee. The general outline of this study includes information on the substrates, the enzyme, and the applications. This review considers the concepts and explores scientific and technological aspects. The “advances” trace the route from the general, molecular, catalytic, and functional information obtained under close to optimal conditions for microbial production through purification, description of the enzyme properties, and the commercial applications to the “perspectives” including expression studies, regulation, and potential uses; aspects related to the progress in our understanding of caffeine biodegradation are also included.

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