Engineering a microbial platform for de novo biosynthesis of diverse methylxanthines

McKeague, Maureen; Wang, Yen-Hsiang; Cravens, Aaron; Win, Maung Nyan; Smolke, Christina D.

doi:10.1016/j.ymben.2016.08.003

Cited by 34 publications

(25 citation statements)

References 55 publications

(76 reference statements)

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“…3-Methylxanthine and other xanthine derivatives have been shown various biomedical effects as adenosine antagonist and inhibitors of Primary Amine Oxidase [57,58]. Besides chemical synthesis, biotransformation and biosynthesis offered alternative way to produce 3-methylxanthine and other xanthine derivatives [22,48]. Due to theophylline degradation characteristic, A. ustus and A. tamarii would be applied in the production of 3-methylxanthine and xanthine with theophylline as feedstock through microbial conversion.…”

Section: Discussionmentioning

confidence: 99%

“…Several xanthine derivatives including 3-methylxanthine have been synthesized chemically for use in medical industry [47]. Except for engineering a microbial platform for de novo biosynthesis of diverse methylxanthins [48], bioconversion from cheaper feedstocks such as caffeine, theophylline and theobromine was an effective pathway to produce high value methylxanthines via metabolically engineered microorganisms [22]. In this study, 3-methylxanthine and xanthine were common and main products in theophylline degradation by A. ustus and A. tamarii.…”

Section: Production Of 3-methylxanthine or Xanthine Through Theophyllmentioning

confidence: 92%

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Isolation, characterization and application of theophylline-degrading Aspergillus fungi

Zhou

Xia

et al. 2020

Microb Cell Fact

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Background: Caffeine, theobromine and theophylline are main purine alkaloid in tea. Theophylline is the downstream metabolite and it remains at a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentation of pu-erh tea through N-demethylation. In this study, tea-derived fungi caused theophylline degradation in the solid-state fermentation. The purpose of this study is identify and isolate theophylline-degrading fungi and investigate their application in production of methylxanthines with theophylline as feedstock through microbial conversion.Results: Seven tea-derived fungi were collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and A. sydowii associated with solid-state fermentation of pu-erh tea have shown ability to degrade theophylline in liquid culture. Particularly, A. ustus and A. tamarii could degrade theophylline highly significantly (p < 0.01). 1,3-dimethyluric acid, 3-methylxanthine, 3-methyluric acid, xanthine and uric acid were detected consecutively by HPLC in A. ustus and A. tamarii, respectively. The data from absolute quantification analysis suggested that 3-methylxanthine and xanthine were the main degraded metabolites in A. ustus and A. tamarii, respectively. 129.48 ± 5.81 mg/L of 3-methylxanthine and 159.11 ± 10.8 mg/L of xanthine were produced by A. ustus and A. tamarii in 300 mg/L of theophylline liquid medium, respectively. Conclusions:For the first time, we confirmed that isolated A. ustus, A. tamarii degrade theophylline through N-demethylation and oxidation. We were able to biologically produce 3-methylxanthine and xanthine efficiently from theophylline through a new microbial synthesis platform with A. ustus and A. tamarii as appropriate starter strains.

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Section: Discussionmentioning

confidence: 99%

Section: Production Of 3-methylxanthine or Xanthine Through Theophyllmentioning

confidence: 92%

Isolation, characterization and application of theophylline-degrading Aspergillus fungi

Zhou

Xia

et al. 2020

Microb Cell Fact

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“…129 In another study, focusing on the synthesis of methylxanthines in S. cerevisiae, higher methylxanthine production was observed when chromosomally introducing expression of biosynthetic enzymes compared with expression from a plasmid. 146 Recently, through testing truncated, weakened promoters to regulate the expression of antibiotic resistance markers, synthetic biologists were able to engineer high-copy plasmids into tunable-copy plasmids regulated by varying antibiotic concentration. 147 This strategy, named Pathway Optimization by Tuning Antibiotic Concentrations (POTAC), was utilized to enhance the titer of lycopene and n-butanol by 10-and 100-fold, respectively (to approximately 11 mg g À1 dry weight lycopene and approximately 110 mg L À1 n-butanol).…”

Section: Strategies For Gene Expression Tuning For Optimizing Phytochmentioning

confidence: 99%

Synthetic biology strategies toward heterologous phytochemical production

Kotopka

Smolke

2018

Nat. Prod. Rep.

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Covering: 2006 to 2018 Phytochemicals are important sources for the discovery and development of agricultural and pharmaceutical compounds, such as pesticides and medicines. However, these compounds are typically present in low abundance in nature, and the biosynthetic pathways for most phytochemicals are not fully elucidated. Heterologous production of phytochemicals in plant, bacterial, and yeast hosts has been pursued as a potential approach to address sourcing issues associated with many valuable phytochemicals, and more recently has been utilized as a tool to aid in the elucidation of plant biosynthetic pathways. Due to the structural complexity of certain phytochemicals and the associated biosynthetic pathways, reconstitution of plant pathways in heterologous hosts can encounter numerous challenges. Synthetic biology approaches have been developed to address these challenges in areas such as precise control over heterologous gene expression, improving functional expression of heterologous enzymes, and modifying central metabolism to increase the supply of precursor compounds into the pathway. These strategies have been applied to advance plant pathway reconstitution and phytochemical production in a wide variety of heterologous hosts. Here, we review synthetic biology strategies that have been recently applied to advance complex phytochemical production in heterologous hosts.

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confidence: 99%

“…Algharrawi et al (2015) reconstructed an engineering Escherichia coli with ndmA and ndmD gene from Pseudomonas putida, capable of producing 3-methylxanthine from exogenously fed theophylline[37] Mckeague et al (2016). engineered the eukaryotic microbial host Saccharomyces cerevisiae for the de novo biosynthesis of methylxanthines[38]. Additionally, 3-methylxanthine was main intermediate metabolite of theobromine through the initial demethylation at position N-7 by theobromine-degrading fungi[32].Because of dominant microbe in the solid-state fermentation causing the significant reduction of caffeine content, Pu-erh tea could be used to select effective stains converting theobromine to 3methylxanthine.…”

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confidence: 99%

3-Methylxanthine production through biodegradation of theobromine by Aspergillus sydowii

Zhou

Zheng

et al. 2020

Preprint

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Background Methylxanthines including caffeine, theobromine and theophylline are natural and synthetic compounds in tea. Theobromine and other methylxanthine could be metabolized by certain kinds of bacteria and fungi. Previous studies confirmed that several isolates from Pu-erh tea could degrade and convert caffeine and theophylline through N-demethylation and related oxidation, respectively. In this study, seven tea-derived fungi were inoculated into various theobromine agar mediums and liquid mediums to assess their capacity in theobromine utilization. Related metabolites with theobromine degradation were detected by using HPLC in the liquid culture inoculated by candidate isolates to investigate potential application in the production of 3-methylxanthine. Results Based on the extent of theobromine utilization, four isolates including Aspergillus niger, Aspergillus sydowii, Aspergillus ustus and Aspergillus tamarii have demonstrated the potential for theobromine biodegradation. Particularly, A. sydowii and A. tamarii could degrade theobromine significantly (p < 0.05) in all given theobromine liquid mediums. 3,7-Dimethyluric acid, 3-methylxanthine, 7-methylxanthine, 3-methyluric acid, xanthine, and uric acid were detected in A. sydowii and A. tamarii culture, respectively, confirming the existence of N-demethylation and oxidation in theobromine catabolism. 3-Methylxanthine was common and main demethylated metabolite of theobromine in A. sydowii and A. tamarii culture. 3-Methylxanthine in A. sydowii culture showed a linear relation with the initial theobromine concentrations that 177.12 ± 14.06 mg/L of 3-methylxanthine was accumulated in TLM-S with 300 mg/L of theobromine. Additionally, pH at 5 and metal ion of Fe 2+ promoted 3-methylxanthine production significantly (p < 0.05). Conclusions This study is the first to confirm that A. sydowii and A. tamarii degrade theobromine through N-demethylation and oxidation, respectively. A. sydowii showed the potential application in 3-methylxanthine production with theobromine as feedstock through the N-demethylation at position N-7.

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Engineering a microbial platform for de novo biosynthesis of diverse methylxanthines

Cited by 34 publications

References 55 publications

Isolation, characterization and application of theophylline-degrading Aspergillus fungi

Isolation, characterization and application of theophylline-degrading Aspergillus fungi

Synthetic biology strategies toward heterologous phytochemical production

3-Methylxanthine production through biodegradation of theobromine by Aspergillus sydowii

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