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 it remains at a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentati...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-03-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12934-020-01333-0 |
| _version_ | 1818067996899278848 |
|---|---|
| author | Binxing Zhou Cunqiang Ma Tao Xia Xiaohong Li Chengqin Zheng Tingting Wu Xiaohui Liu |
| author_facet | Binxing Zhou Cunqiang Ma Tao Xia Xiaohong Li Chengqin Zheng Tingting Wu Xiaohui Liu |
| author_sort | Binxing Zhou |
| collection | DOAJ |
| description | 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 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. |
| first_indexed | 2024-12-10T15:32:34Z |
| format | Article |
| id | doaj.art-fce5836a517d48bcbeb147a5e63a8a4d |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-10T15:32:34Z |
| publishDate | 2020-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-fce5836a517d48bcbeb147a5e63a8a4d2022-12-22T01:43:20ZengBMCMicrobial Cell Factories1475-28592020-03-0119111310.1186/s12934-020-01333-0Isolation, characterization and application of theophylline-degrading Aspergillus fungiBinxing Zhou0Cunqiang Ma1Tao Xia2Xiaohong Li3Chengqin Zheng4Tingting Wu5Xiaohui Liu6Long Run Pu-erh Tea College, Yunnan Agricultural UniversityLong Run Pu-erh Tea College, Yunnan Agricultural UniversityState Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural UniversityLong Run Pu-erh Tea College, Yunnan Agricultural UniversityLong Run Pu-erh Tea College, Yunnan Agricultural UniversityLong Run Pu-erh Tea College, Yunnan Agricultural UniversityLong Run Pu-erh Tea College, Yunnan Agricultural UniversityAbstract 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.http://link.springer.com/article/10.1186/s12934-020-01333-0AspergillusTheophylline3-MethylxanthineBioconversionTeaPathway |
| spellingShingle | Binxing Zhou Cunqiang Ma Tao Xia Xiaohong Li Chengqin Zheng Tingting Wu Xiaohui Liu Isolation, characterization and application of theophylline-degrading Aspergillus fungi Microbial Cell Factories Aspergillus Theophylline 3-Methylxanthine Bioconversion Tea Pathway |
| title | Isolation, characterization and application of theophylline-degrading Aspergillus fungi |
| title_full | Isolation, characterization and application of theophylline-degrading Aspergillus fungi |
| title_fullStr | Isolation, characterization and application of theophylline-degrading Aspergillus fungi |
| title_full_unstemmed | Isolation, characterization and application of theophylline-degrading Aspergillus fungi |
| title_short | Isolation, characterization and application of theophylline-degrading Aspergillus fungi |
| title_sort | isolation characterization and application of theophylline degrading aspergillus fungi |
| topic | Aspergillus Theophylline 3-Methylxanthine Bioconversion Tea Pathway |
| url | http://link.springer.com/article/10.1186/s12934-020-01333-0 |
| work_keys_str_mv | AT binxingzhou isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT cunqiangma isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT taoxia isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT xiaohongli isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT chengqinzheng isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT tingtingwu isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi AT xiaohuiliu isolationcharacterizationandapplicationoftheophyllinedegradingaspergillusfungi |