10.1021/acs.molpharmaceut.5b00954.s001 Shinichi Ikushiro Shinichi Ikushiro Miyu Nishikawa Miyu Nishikawa Yuuka Masuyama Yuuka Masuyama Tadashi Shouji Tadashi Shouji Miharu Fujii Miharu Fujii Masahiro Hamada Masahiro Hamada Noriyuki Nakajima Noriyuki Nakajima Moshe Finel Moshe Finel Kaori Yasuda Kaori Yasuda Masaki Kamakura Masaki Kamakura Toshiyuki Sakaki Toshiyuki Sakaki Biosynthesis of Drug Glucuronide Metabolites in the Budding Yeast <i>Saccharomyces cerevisiae</i> American Chemical Society 2016 compound Budding Yeast Saccharomyces cerevisiae Glucuronidation Drug Glucuronide Metabolites acid acyl glucuronides UGT coexpression system 1A yeast cells coexpressing UGDH 2016-05-30 00:00:00 Journal contribution https://acs.figshare.com/articles/journal_contribution/Biosynthesis_of_Drug_Glucuronide_Metabolites_in_the_Budding_Yeast_i_Saccharomyces_cerevisiae_i_/3423327 Glucuronidation is one of the most common pathways in mammals for detoxification and elimination of hydrophobic xenobiotic compounds, including many drugs. Metabolites, however, can form active or toxic compounds, such as acyl glucuronides, and their safety assessment is often needed. The absence of efficient means for <i>in vitro</i> synthesis of correct glucuronide metabolites frequently limits such toxicological analyses. To overcome this hurdle we have developed a new approach, the essence of which is a coexpression system containing a human, or another mammalian UDP-glucuronosyltransferases (UGTs), as well as UDP-glucose-6-dehydrogenase (UGDH), within the budding yeast, <i>Saccharomyces cerevisiae</i>. The system was first tested using resting yeast cells coexpressing UGDH and human UGT1A6, 7-hydroxycoumarin as the substrate, in a reaction medium containing 8% glucose, serving as a source of UDP-glucuronic acid. Glucuronides were readily formed and recovered from the medium. Subsequently, by selecting suitable mammalian UGT enzyme for the coexpression system we could obtain the desired glucuronides of various compounds, including molecules with multiple conjugation sites and acyl glucuronides of several carboxylic acid containing drugs, namely, mefenamic acid, flufenamic acid, and zomepirac. In conclusion, a new and flexible yeast system with mammalian UGTs has been developed that exhibits a capacity for efficient production of various glucuronides, including acyl glucuronides.