Metabolic Engineering of Saccharomyces cerevisiae for Caffeine and Theobromine Production |
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| Authors: | Lu Jin Mohammad Wadud Bhuiya Mengmeng Li XiangQi Liu Jixiang Han WeiWei Deng Min Wang Oliver Yu Zhengzhu Zhang |
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| Institution: | 1. Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education, Anhui Agricultural University, Hefei, PR China.; 2. Conagen Inc., St. Louis, Missouri, United States of America.; 3. Donald Danforth Plant Science Center, St. Louis, Missouri, United States of America.; University of Geneva, Switzerland, |
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| Abstract: | Caffeine (1, 3, 7-trimethylxanthine) and theobromine (3, 7-dimethylxanthine) are the major purine alkaloids in plants, e.g. tea (Camellia sinensis) and coffee (Coffea arabica). Caffeine is a major component of coffee and is used widely in food and beverage industries. Most of the enzymes involved in the caffeine biosynthetic pathway have been reported previously. Here, we demonstrated the biosynthesis of caffeine (0.38 mg/L) by co-expression of Coffea arabica xanthosine methyltransferase (CaXMT) and Camellia sinensis caffeine synthase (TCS) in Saccharomyces cerevisiae. Furthermore, we endeavored to develop this production platform for making other purine-based alkaloids. To increase the catalytic activity of TCS in an effort to increase theobromine production, we identified four amino acid residues based on structural analyses of 3D-model of TCS. Two TCS1 mutants (Val317Met and Phe217Trp) slightly increased in theobromine accumulation and simultaneously decreased in caffeine production. The application and further optimization of this biosynthetic platform are discussed. |
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