title: Biosynthesis of Drug Glucuronide Metabolites in the Budding Yeast Saccharomyces cerevisiae.
abstract: 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 in vitro 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, Saccharomyces cerevisiae. 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.
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Ikushiro
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.|Imizu Institute, TOPU BIO RESEARCH Co., Ltd , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Miyu
authors.initials: M
authors.lastname: Nishikawa
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Yuuka
authors.initials: Y
authors.lastname: Masuyama
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Shouji
authors.affiliation: Imizu Institute, TOPU BIO RESEARCH Co., Ltd , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Miharu
authors.initials: M
authors.lastname: Fujii
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hamada
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nakajima
authors.affiliation: Division of Pharmaceutical Chemistry and Technology, University of Helsinki , P.O. Box 56, 00014 Helsinki, Finland.
authors.forename: Moshe
authors.initials: M
authors.lastname: Finel
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Yasuda
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kamakura
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.|Imizu Institute, TOPU BIO RESEARCH Co., Ltd , 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sakaki
mesh_terms: D000818:Animals; D002264:Carboxylic Acids / Q000378:metabolism; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000378:metabolism; D006801:Humans; D010088:Oxidoreductases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D004718:Saccharomycetales / Q000378:metabolism*; D014468:Umbelliferones / Q000378:metabolism
chemical_list: D002264:Carboxylic Acids; D020719:Glucuronides; D014468:Umbelliferones; C031477:7-hydroxycoumarin; D010088:Oxidoreductases; C110655:UDP-glucuronosyltransferase, UGT1A6; D014453:Glucuronosyltransferase
keywords: UDP-glucose-6-dehydrogenase; UDP-glucuronic acid; UDP-glucuronosyltransferase; acyl glucuronide; budding yeast