Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production

• Published in: PloS one 2014-08, Vol.9 (8), p.e105368-e105368
• Main Authors: Jin, Lu, Bhuiya, Mohammad Wadud, Li, Mengmeng, Liu, XiangQi, Han, Jixiang, Deng, WeiWei, Wang, Min, Yu, Oliver, Zhang, Zhengzhu
• Format: Article
• Language: English
• Subjects: Alkaloids; Amino acids; Analysis; Beverage industry; Biochemistry; Biology and Life Sciences; Biosynthesis; Biosynthetic Pathways; Biotechnology; Caffeine; Caffeine - genetics; Caffeine - metabolism; Caffeine synthase; Camellia sinensis; Camellia sinensis - enzymology; Catalysis; Catalytic activity; Catechin; Cloning; Coffea - enzymology; Coffea arabica; Coffee; E coli; Education; Engineering; Engineering and Technology; Enzymes; Food industry; Genomics; Industrial Microbiology - methods; Laboratories; Metabolic engineering; Metabolic Engineering - methods; Metabolism; Methyltransferase; Methyltransferases - genetics; Methyltransferases - metabolism; Mutants; Mutation; Optimization; Physiological aspects; Plant sciences; Plasmids; Proteins; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Tea; Theobromine - genetics; Theobromine - metabolism; Three dimensional models; Transferases; Yeast
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0105368

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.