Aromadendrin is an important component of citrus flavonoids, and flavanone-3-hydroxylase (F3H) plays a key role in the formation pathway of aromadendrin. In this study, we summarized and analyzed the characteristics of F3H, and expressed them in Escherichia coli BL21. First, 5 kinds of f3h from Citrus reticulata Banco, Gossypium hirsutum, Zea mays, Saussurea medusa and Petroselinum crispumin NCBI database were chosen. Then, physicochemical properties, secondary and tertiary structure of protein, sequence alignment and other bioinformatics analysis were carried out. At last, recombinant expression system were constructed and expressed with E.coli BL21. The open reading frame of f3h gene was 1032-1119bp, encoding 343-372 amino acids. The molecular weight of F3H was 38.89-41.45kDa, and the theoretical isoelectric point was 5.43-5.96. The physical and chemical properties, domains and protein structures of F3H from different species were also similar.By constructing the expression system of E.coli BL21,we found that all F3H could be prokaryotic expression. Except the F3H from Saussurea medusi, the other 4 kinds of F3H could convert naringin to aromadendrin with different production amounts, and the highest production amount of ZmF3H was 20.12mg/L.