In recent decades, biocatalysis has gained the prominence in the chemical industry, particularly due to its excellent enantio-, regio- and chemoselectivity. There is a high demand for asymmetric alkene reductase, as it could generate up to two chiral centers. Among these enzymes, the ‘old yellow enzyme’ (OYE) has been the most extensively investigated and industrially used. In contrast with OYE using the universal flavin cofactors, F420H2-Dependent Reductases (FDR) use unusual deazaflavin cofactor F420 which is found only in restricted organisms and has not been extensively studied so far. Given the similar reactivity profile of OYEs and FDRs, FDRs are also could be a potential candidate for useful biotransformation, while the unique properties of F420 might lead to better activities or different substrate scope. In this study, typical substrates of OYE and a couple of compounds of our interest are tested to investigate substrate range and enantioselectivity of FDRs. Interestingly, FDRs tend to show different enantioselectivity from OYEs.