A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials
2,3,5,6-Tetramethylpyrazine (TMP) is an active pharmaceutical ingredient originally isolated from Ligusticum wallichii for curing cardiovascular and cerebrovascular diseases and is widely used as a popular flavoring additive in the food industry. Hence, there is a great interest in developing new st...
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| Format: | Article |
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2021.792023/full |
| _version_ | 1798025177864339456 |
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| author | Jing Li Jian Lu Zhilin Ma Jianxiu Li Xianrui Chen Mengxue Diao Nengzhong Xie |
| author_facet | Jing Li Jian Lu Zhilin Ma Jianxiu Li Xianrui Chen Mengxue Diao Nengzhong Xie |
| author_sort | Jing Li |
| collection | DOAJ |
| description | 2,3,5,6-Tetramethylpyrazine (TMP) is an active pharmaceutical ingredient originally isolated from Ligusticum wallichii for curing cardiovascular and cerebrovascular diseases and is widely used as a popular flavoring additive in the food industry. Hence, there is a great interest in developing new strategies to produce this high-value compound in an ecological and economical way. Herein, a cost-competitive combinational approach was proposed to accomplish green and high-efficiency production of TMP. First, microbial cell factories were constructed to produce acetoin (3-hydroxy-2-butanone, AC), an endogenous precursor of TMP, by introducing a biosynthesis pathway coupled with an intracellular NAD+ regeneration system to the wild-type Escherichia coli. To further improve the production of (R)-AC, the metabolic pathways of by-products were impaired or blocked stepwise by gene manipulation, resulting in 40.84 g/L (R)-AC with a high optical purity of 99.42% in shake flasks. Thereafter, an optimal strain designated GXASR11 was used to convert the hydrolysates of inexpensive feedstocks into (R)-AC and achieved a titer of 86.04 g/L within 48 h in a 5-L fermenter under optimized fermentation conditions. To the best of our knowledge, this is the highest (R)-AC production with high optical purity (≥98%) produced from non-food raw materials using recombinant E. coli. The supernatant of fermentation broth was mixed with diammonium phosphate (DAP) to make a total volume of 20 ml and transferred to a high-pressure microreactor. Finally, 56.72 g/L TMP was obtained in 3 h via the condensation reaction with a high conversion rate (85.30%) under optimal reaction conditions. These results demonstrated a green and sustainable approach to efficiently produce high-valued TMP, which realized value addition of low-cost renewables. |
| first_indexed | 2024-04-11T18:14:34Z |
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| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-11T18:14:34Z |
| publishDate | 2022-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-c2a230457968452fb19dda8568b541c22022-12-22T04:09:59ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-01-01910.3389/fbioe.2021.792023792023A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw MaterialsJing Li0Jian Lu1Zhilin Ma2Jianxiu Li3Xianrui Chen4Mengxue Diao5Nengzhong Xie6Life Science and Technology College, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, Guangxi, ChinaLife Science and Technology College, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, Guangxi, ChinaLife Science and Technology College, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, Guangxi, ChinaState Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Key Laboratory of Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, ChinaState Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Key Laboratory of Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, ChinaState Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Key Laboratory of Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, ChinaState Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Key Laboratory of Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, China2,3,5,6-Tetramethylpyrazine (TMP) is an active pharmaceutical ingredient originally isolated from Ligusticum wallichii for curing cardiovascular and cerebrovascular diseases and is widely used as a popular flavoring additive in the food industry. Hence, there is a great interest in developing new strategies to produce this high-value compound in an ecological and economical way. Herein, a cost-competitive combinational approach was proposed to accomplish green and high-efficiency production of TMP. First, microbial cell factories were constructed to produce acetoin (3-hydroxy-2-butanone, AC), an endogenous precursor of TMP, by introducing a biosynthesis pathway coupled with an intracellular NAD+ regeneration system to the wild-type Escherichia coli. To further improve the production of (R)-AC, the metabolic pathways of by-products were impaired or blocked stepwise by gene manipulation, resulting in 40.84 g/L (R)-AC with a high optical purity of 99.42% in shake flasks. Thereafter, an optimal strain designated GXASR11 was used to convert the hydrolysates of inexpensive feedstocks into (R)-AC and achieved a titer of 86.04 g/L within 48 h in a 5-L fermenter under optimized fermentation conditions. To the best of our knowledge, this is the highest (R)-AC production with high optical purity (≥98%) produced from non-food raw materials using recombinant E. coli. The supernatant of fermentation broth was mixed with diammonium phosphate (DAP) to make a total volume of 20 ml and transferred to a high-pressure microreactor. Finally, 56.72 g/L TMP was obtained in 3 h via the condensation reaction with a high conversion rate (85.30%) under optimal reaction conditions. These results demonstrated a green and sustainable approach to efficiently produce high-valued TMP, which realized value addition of low-cost renewables.https://www.frontiersin.org/articles/10.3389/fbioe.2021.792023/fulltetramethylpyrazinenon-food raw materialssynthetic biologyacetoinmetabolic engineeringgreen process |
| spellingShingle | Jing Li Jian Lu Zhilin Ma Jianxiu Li Xianrui Chen Mengxue Diao Nengzhong Xie A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials Frontiers in Bioengineering and Biotechnology tetramethylpyrazine non-food raw materials synthetic biology acetoin metabolic engineering green process |
| title | A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials |
| title_full | A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials |
| title_fullStr | A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials |
| title_full_unstemmed | A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials |
| title_short | A Green Route for High-Yield Production of Tetramethylpyrazine From Non-Food Raw Materials |
| title_sort | green route for high yield production of tetramethylpyrazine from non food raw materials |
| topic | tetramethylpyrazine non-food raw materials synthetic biology acetoin metabolic engineering green process |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2021.792023/full |
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