Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production
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 i...
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Published in | PloS one Vol. 9; no. 8; p. e105368 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Public Library of Science
18.08.2014
Public Library of Science (PLoS) |
Subjects | |
Online Access | Get full text |
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Summary: | 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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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 SC0001295 USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22) Conceived and designed the experiments: ZZZ OY. Performed the experiments: LJ MWB MML XQL JXH MW. Analyzed the data: LJ MWB MML XQL JXH MW. Contributed reagents/materials/analysis tools: LJ MWB. Wrote the paper: LJ MWB WWD ZZZ OY. Competing Interests: The authors have the following interests. M. W. Bhuiya and J. X. Han are employed by Conagen Inc. This does not alter the authors’ adherence to PLOS ONE policies on sharing data and materials. |
ISSN: | 1932-6203 1932-6203 |
DOI: | 10.1371/journal.pone.0105368 |