The invention provides a bioenzyme-containing diesel oil booster composition, comprising a cetane number improver and a bioenzyme at a weight ratio of 1:0.01-1; the bioenzyme is sulfonase, alcohol oxidase, aldehyde Oxidase, amine oxidase, sulfhydryl oxidase, methylmercaptan oxidase, menaquinone oxidase, peroxidase, ferrochelatase, decarboxylase, aldolase, oxoacid lyase, demethylation Mixture of enzymes, alcohol, acid, ester dehydratase, desulfhydrylase, desulfomethylase, cobalt chelatase, magnesium chelatase. A bio-enzyme-containing diesel oil booster composition of the present invention can realize high-speed catalytic performance in a mild environment by utilizing the characteristics of a bio-enzyme, and is used in combination with the existing cetane number improver, The catalytic combustion efficiency of the cetane number improver in diesel is significantly improved, and the combustion performance of diesel is further improved.