In the research and development of new drugs, it is very important to investigate the in vitrometabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant humandrug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major humanUDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from humanlivercDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insectcell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insectcells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, 1H NMR, and 13C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insectcell expression systems constructed in this study can be used for in vitrometabolism evaluation in the early stage of new drugsresearch and development, and also provide a new tool for the synthesis of glucuronide metabolites.