Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine
Abstract Background 7-Methylxanthine, a derivative of caffeine noted for its lack of toxicity and ability to treat and even prevent myopia progression, is a high-value biochemical with limited natural availability. Attempts to produce 7-methylxanthine through purely chemical methods of synthesis are...
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| Format: | Article |
| Language: | English |
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BMC
2023-01-01
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| Series: | Journal of Biological Engineering |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13036-022-00316-6 |
| _version_ | 1797952497937022976 |
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| author | Meredith B. Mock Ryan M. Summers |
| author_facet | Meredith B. Mock Ryan M. Summers |
| author_sort | Meredith B. Mock |
| collection | DOAJ |
| description | Abstract Background 7-Methylxanthine, a derivative of caffeine noted for its lack of toxicity and ability to treat and even prevent myopia progression, is a high-value biochemical with limited natural availability. Attempts to produce 7-methylxanthine through purely chemical methods of synthesis are faced with complicated chemical processes and/or the requirement of a variety of hazardous chemicals, resulting in low yields and racemic mixtures of products. In recent years, we have developed engineered microbial cells to produce several methylxanthines, including 3-methylxanthine, theobromine, and paraxanthine. The purpose of this study is to establish a more efficient biosynthetic process for the production of 7-methylxanthine from caffeine. Results Here, we describe the use of a mixed-culture system composed of Escherichia coli strains engineered as caffeine and theobromine “specialist” cells. Optimal reaction conditions for the maximal conversion of caffeine to 7-methylxanthine were determined to be equal concentrations of caffeine and theobromine specialist cells at an optical density (600 nm) of 50 reacted with 2.5 mM caffeine for 5 h. When scaled-up to 560 mL, the simple biocatalytic reaction produced 183.81 mg 7-methylxanthine from 238.38 mg caffeine under ambient conditions, an 85.6% molar conversion. Following HPLC purification and solvent evaporation, 153.3 mg of dried 7-methylxanthine powder was collected, resulting in an 83.4% product recovery. Conclusion We present the first report of a biocatalytic process designed specifically for the production and purification of the high-value biochemical 7-methylxanthine from caffeine using a mixed culture of E. coli strains. This process constitutes the most efficient method for the production of 7-methylxanthine from caffeine to date. |
| first_indexed | 2024-04-10T22:47:18Z |
| format | Article |
| id | doaj.art-96e4b9e9f8fd415da5ebfb0d94a601e3 |
| institution | Directory Open Access Journal |
| issn | 1754-1611 |
| language | English |
| last_indexed | 2024-04-10T22:47:18Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biological Engineering |
| spelling | doaj.art-96e4b9e9f8fd415da5ebfb0d94a601e32023-01-15T12:14:07ZengBMCJournal of Biological Engineering1754-16112023-01-0117111110.1186/s13036-022-00316-6Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthineMeredith B. Mock0Ryan M. Summers1Department of Chemical and Biological Engineering, The University of AlabamaDepartment of Chemical and Biological Engineering, The University of AlabamaAbstract Background 7-Methylxanthine, a derivative of caffeine noted for its lack of toxicity and ability to treat and even prevent myopia progression, is a high-value biochemical with limited natural availability. Attempts to produce 7-methylxanthine through purely chemical methods of synthesis are faced with complicated chemical processes and/or the requirement of a variety of hazardous chemicals, resulting in low yields and racemic mixtures of products. In recent years, we have developed engineered microbial cells to produce several methylxanthines, including 3-methylxanthine, theobromine, and paraxanthine. The purpose of this study is to establish a more efficient biosynthetic process for the production of 7-methylxanthine from caffeine. Results Here, we describe the use of a mixed-culture system composed of Escherichia coli strains engineered as caffeine and theobromine “specialist” cells. Optimal reaction conditions for the maximal conversion of caffeine to 7-methylxanthine were determined to be equal concentrations of caffeine and theobromine specialist cells at an optical density (600 nm) of 50 reacted with 2.5 mM caffeine for 5 h. When scaled-up to 560 mL, the simple biocatalytic reaction produced 183.81 mg 7-methylxanthine from 238.38 mg caffeine under ambient conditions, an 85.6% molar conversion. Following HPLC purification and solvent evaporation, 153.3 mg of dried 7-methylxanthine powder was collected, resulting in an 83.4% product recovery. Conclusion We present the first report of a biocatalytic process designed specifically for the production and purification of the high-value biochemical 7-methylxanthine from caffeine using a mixed culture of E. coli strains. This process constitutes the most efficient method for the production of 7-methylxanthine from caffeine to date.https://doi.org/10.1186/s13036-022-00316-67-methylxanthineCaffeineBiocatalysisN-demethylaseMixed-culture |
| spellingShingle | Meredith B. Mock Ryan M. Summers Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine Journal of Biological Engineering 7-methylxanthine Caffeine Biocatalysis N-demethylase Mixed-culture |
| title | Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine |
| title_full | Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine |
| title_fullStr | Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine |
| title_full_unstemmed | Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine |
| title_short | Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine |
| title_sort | mixed culture biocatalytic production of the high value biochemical 7 methylxanthine |
| topic | 7-methylxanthine Caffeine Biocatalysis N-demethylase Mixed-culture |
| url | https://doi.org/10.1186/s13036-022-00316-6 |
| work_keys_str_mv | AT meredithbmock mixedculturebiocatalyticproductionofthehighvaluebiochemical7methylxanthine AT ryanmsummers mixedculturebiocatalyticproductionofthehighvaluebiochemical7methylxanthine |