Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine

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dc.contributor.authorMock, Meredith B.
dc.contributor.authorSummers, Ryan M.
dc.contributor.otherUniversity of Alabama Tuscaloosa
dc.date.accessioned2023-09-28T19:15:39Z
dc.date.available2023-09-28T19:15:39Z
dc.date.issued2023
dc.description.abstractBackground 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.en_US
dc.format.mediumelectronic
dc.format.mimetypeapplication/pdf
dc.identifier.citationMock, M. B., & Summers, R. M. (2023). Mixed culture biocatalytic production of the high-value biochemical 7-methylxanthine. In Journal of Biological Engineering (Vol. 17, Issue 1). Springer Science and Business Media LLC. https://doi.org/10.1186/s13036-022-00316-6
dc.identifier.doi10.1186/s13036-022-00316-6
dc.identifier.orcidhttps://orcid.org/0000-0002-5316-1575
dc.identifier.urihttps://ir.ua.edu/handle/123456789/11194
dc.languageEnglish
dc.language.isoen_US
dc.publisherBMC
dc.rights.licenseAttribution 4.0 International (CC BY 4.0)
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subject7-methylxanthine
dc.subjectCaffeine
dc.subjectBiocatalysis
dc.subjectN-demethylase
dc.subjectMixed-culture
dc.subjectFORM DEPRIVATION MYOPIA
dc.subjectCAFFEINE DEMETHYLASE
dc.subjectFERMENTATION
dc.subjectDEGRADATION
dc.subjectXANTHINES
dc.subjectBiochemical Research Methods
dc.subjectBiotechnology & Applied Microbiology
dc.titleMixed culture biocatalytic production of the high-value biochemical 7-methylxanthineen_US
dc.typeArticle
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