Isolation, characterization and application of theophylline-degrading Aspergillus fungi

Abstract Background: Caffeine, theobromine and theophylline are main purine alkaloid in tea. Theophylline is the downstream metabolite and stays a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentation of tea through N-demethylation. In this study, tea-derived fungi caused theophylline degradation in the solid-state fermentation. The aim of this study was isolate theophylline-degrading fungi and to investigate their application in production of methylxanthines with theophylline as feedstock through microbial conversion. Results: Of seven tea-derived fungi collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and Aspergillus sydowii associated with solid-state fermentation of pu-erh tea have shown ability to degrade theophylline. A. ustus and A. tamarii could degrade theophylline highly significantly (p < 0.01) in liquid culture. 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 suggest that 3-methylxanthine and xanthine were 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 respectively in 300 mg/L of theophylline liquid medium. Conclusions: For the first time, we confirmed that isolated Aspergillus ustus, Aspergillus 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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Isolation, characterization and application of theophylline-degrading Aspergillus fungi

10.21203/rs.2.18212/v2 ◽

2020 ◽

Author(s):

Bin-Xing ZHOU ◽

Cunqiang Ma ◽

Tao Xia ◽

Xiaohong Li ◽

Chengqin Zheng ◽

...

Keyword(s):

Solid State ◽

Solid State Fermentation ◽

Liquid Culture ◽

Absolute Quantification ◽

Aspergillus Sydowii ◽

Microbial Conversion ◽

Aspergillus Tamarii ◽

Aspergillus Ustus ◽

Quantification Analysis ◽

First Time

Abstract 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:Sseven tea-derived fungi were collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and Aspergillus 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 Aspergillus ustus, Aspergillus 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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3-Methylxanthine production through biodegradation of theobromine by Aspergillus sydowii PT-2

10.21203/rs.3.rs-19407/v2 ◽

2020 ◽

Author(s):

bin-xing ZHOU ◽

Cunqiang Ma ◽

Chengqin Zheng ◽

Tao Xia ◽

Bingsong Ma ◽

...

Keyword(s):

Uric Acid ◽

Potential Application ◽

Metal Ion ◽

Liquid Culture ◽

Aspergillus Sydowii ◽

Aspergillus Tamarii ◽

Synthetic Compounds ◽

Aspergillus Ustus ◽

Bacteria And Fungi ◽

Microbial Isolates

Abstract Background: Methylxanthines, including caffeine, theobromine and theophylline, are natural and synthetic compounds in tea, which could be metabolized by certain kinds of bacteria and fungi. Previous studies confirmed that several microbial isolates from Pu-erh tea could degrade and convert caffeine and theophylline. We speculated that these candidate isolates also could degrade and convert theobromine through N-demethylation and oxidation. In this study, seven tea-derived fungal strains were inoculated into various theobromine agar medias and theobromine liquid mediums to assess their capacity in theobromine utilization. Related metabolites with theobromine degradation were detected by using HPLC in the liquid culture to investigate their potential application in the production of 3-methylxanthine. Results: Based on theobromine utilization capacity, Aspergillus niger PT-1, Aspergillus sydowii PT-2, Aspergillus ustus PT-6 and Aspergillus tamarii PT-7 have demonstrated the potential for theobromine biodegradation. Particularly, A. sydowii PT-2 and A. tamarii PT-7 could degrade theobromine significantly (p < 0.05) in all given liquid mediums. 3,7-Dimethyluric acid, 3-methylxanthine, 7-methylxanthine, 3-methyluric acid, xanthine, and uric acid were detected in A. sydowii PT-2 and A. tamarii PT-7 culture, respectively, which confirmed the existence of N-demethylation and oxidation in theobromine catabolism. 3-Methylxanthine was common and main demethylated metabolite of theobromine in the liquid culture. 3-Methylxanthine in A. sydowii PT-2 culture showed a linear relation with initial theobromine concentrations that 177.12 ± 14.06 mg/L 3-methylxanthine was accumulated in TLM-S with 300 mg/L theobromine. Additionally, pH at 5 and metal ion of Fe2+ promoted 3-methylxanthine production significantly (p < 0.05). Conclusions: This study is the first to confirm that A. sydowii PT-2 and A. tamarii PT-7 degrade theobromine through N-demethylation and oxidation, respectively. A. sydowii PT-2 showed the potential application in 3-methylxanthine production with theobromine as feedstock through the N-demethylation at N-7 position.

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3-Methylxanthine production through biodegradation of theobromine by Aspergillus sydowii

10.21203/rs.3.rs-19407/v1 ◽

2020 ◽

Author(s):

Bin-Xing ZHOU ◽

Cunqiang Ma ◽

Chengqin Zheng ◽

Tao Xia ◽

Binsong Ma ◽

...

Keyword(s):

Uric Acid ◽

Aspergillus Niger ◽

Potential Application ◽

Metal Ion ◽

Liquid Culture ◽

Aspergillus Sydowii ◽

Aspergillus Tamarii ◽

Synthetic Compounds ◽

Aspergillus Ustus ◽

Bacteria And Fungi

Abstract 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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Production of β-fructofuranosidase with transfructosylating activity by Aspergillus tamarii URM4634 Solid-State Fermentation on agroindustrial by-products

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2019.12.084 ◽

2020 ◽

Vol 144 ◽

pp. 343-350 ◽

Cited By ~ 2

Author(s):

Rodrigo Lira de Oliveira ◽

Marcos Fellipe da Silva ◽

Attilio Converti ◽

Tatiana Souza Porto

Keyword(s):

Solid State ◽

Solid State Fermentation ◽

Aspergillus Tamarii ◽

By Products

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Kinetic Study of Glucosamine Production Using Aspergillus sydowii BCRC 31742 under Solid-State Fermentation

Molecules ◽

10.3390/molecules25204832 ◽

2020 ◽

Vol 25 (20) ◽

pp. 4832

Author(s):

Jia Wei Peng ◽

Ho Shing Wu

Keyword(s):

Solid State ◽

Moisture Content ◽

Solid State Fermentation ◽

Low Cost ◽

High Yield ◽

Aspergillus Sydowii ◽

Fermentation Conditions ◽

High Productivity ◽

State Culture ◽

Solid State Culture

In the present study, we aimed to obtain a high yield and productivity for glucosamine using a low-cost solid-state culture with Aspergillus sydowii BCRC 31742. The fermentation conditions, such as inoculum biomass, moisture content, and supplemental volume and mineral salt, were chosen to achieve high productivity of glucosamine (GlcN). When the initial supplemental volume used was 3 mL/g substrate, the yield and productivity of GlcN were 48.7 mg/gds and 0.69 mg/gds·h, respectively. This result will be helpful for the industrialization of the process.

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