Protein engineering of microbial cholesterol oxidases: a molecular approach toward development of new enzymes with new properties

Cholesterol oxidase, a flavoenzyme, catalyzes two
reactions in one active site: oxidation and isomerization. This
enzyme has been isolated from a variety of microorganisms,
mostly from actinomycetes. This enzyme has been widely
used in clinical laboratories for cholesterol assays and was
subsequently determined to have other potential applications.
Engineering of cholesterol oxidase have enabled the identification of critical residues, and the information derived could
lead to the rational development of improved types of the
enzyme with increased stability and better functional properties. This review is the first that exclusively summarizes the
reported results on the engineering of bacterial cholesterol
oxidases aimed at improving their thermal and chemical stability, catalytic activity, and substrate specificity