Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions
(R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine, a key chiral intermediate of selective tetrodotoxin-sensitive blockers, was efficiently synthesized by a bienzyme cascade system formed by with R-ω-transaminase (ATA117) and an alcohol dehydrogenase (ADH) co-expression system. Herein, we report that...
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MDPI AG
2022-10-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/21/7331 |
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| author | Yuan Lu Jinmei Wang Haobo Xu Chuyue Zhang Pengpeng Cheng Lihua Du Lan Tang Jinghua Li Zhimin Ou |
| author_facet | Yuan Lu Jinmei Wang Haobo Xu Chuyue Zhang Pengpeng Cheng Lihua Du Lan Tang Jinghua Li Zhimin Ou |
| author_sort | Yuan Lu |
| collection | DOAJ |
| description | (R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine, a key chiral intermediate of selective tetrodotoxin-sensitive blockers, was efficiently synthesized by a bienzyme cascade system formed by with R-ω-transaminase (ATA117) and an alcohol dehydrogenase (ADH) co-expression system. Herein, we report that the use of ATA117 as the biocatalyst for the amination of 3,5-bistrifluoromethylacetophenone led to the highest efficiency in product performance (enantiomeric excess > 99.9%). Moreover, to further improve the product yield, ADH was introduced into the reaction system to promote an equilibrium shift. Additionally, bienzyme cascade system was constructed by five different expression systems, including two tandem expression recombinant plasmids (pETDuet-ATA117-ADH and pACYCDuet-ATA117-ADH) and three co-expressed dual-plasmids (pETDuet-ATA117/pET28a-ADH, pACYCDuet-ATA117/pET28a-ADH, and pACYCDuet-ATA117/pETDuet-ADH), utilizing recombinant engineered bacteria. Subsequent studies revealed that as compared with ATA117 single enzyme, the substrate handling capacity of BL21(DE3)/pETDuet-ATA117-ADH (0.25 g wet weight) developed for bienzyme cascade system was increased by 1.50 folds under the condition of 40 °C, 180 rpm, 0.1 M pH9 Tris-HCl for 24 h. To the best of our knowledge, ours is the first report demonstrating the production of (R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine using a bienzyme cascade system, thus providing valuable insights into the biosynthesis of chiral amines. |
| first_indexed | 2024-03-09T18:47:37Z |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T18:47:37Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj.art-0b7c3d1a876e4182b5b35c21bd6674e32023-11-24T06:02:37ZengMDPI AGMolecules1420-30492022-10-012721733110.3390/molecules27217331Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase FunctionsYuan Lu0Jinmei Wang1Haobo Xu2Chuyue Zhang3Pengpeng Cheng4Lihua Du5Lan Tang6Jinghua Li7Zhimin Ou8College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, ChinaCollege of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, China(R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine, a key chiral intermediate of selective tetrodotoxin-sensitive blockers, was efficiently synthesized by a bienzyme cascade system formed by with R-ω-transaminase (ATA117) and an alcohol dehydrogenase (ADH) co-expression system. Herein, we report that the use of ATA117 as the biocatalyst for the amination of 3,5-bistrifluoromethylacetophenone led to the highest efficiency in product performance (enantiomeric excess > 99.9%). Moreover, to further improve the product yield, ADH was introduced into the reaction system to promote an equilibrium shift. Additionally, bienzyme cascade system was constructed by five different expression systems, including two tandem expression recombinant plasmids (pETDuet-ATA117-ADH and pACYCDuet-ATA117-ADH) and three co-expressed dual-plasmids (pETDuet-ATA117/pET28a-ADH, pACYCDuet-ATA117/pET28a-ADH, and pACYCDuet-ATA117/pETDuet-ADH), utilizing recombinant engineered bacteria. Subsequent studies revealed that as compared with ATA117 single enzyme, the substrate handling capacity of BL21(DE3)/pETDuet-ATA117-ADH (0.25 g wet weight) developed for bienzyme cascade system was increased by 1.50 folds under the condition of 40 °C, 180 rpm, 0.1 M pH9 Tris-HCl for 24 h. To the best of our knowledge, ours is the first report demonstrating the production of (R)-1-[3,5-bis(trifluoromethyl)phenyl]ethanamine using a bienzyme cascade system, thus providing valuable insights into the biosynthesis of chiral amines.https://www.mdpi.com/1420-3049/27/21/7331alcohol dehydrogenasebienzyme cascade systemchiral aminesR-ω-transaminaserecombinant engineered bacteria |
| spellingShingle | Yuan Lu Jinmei Wang Haobo Xu Chuyue Zhang Pengpeng Cheng Lihua Du Lan Tang Jinghua Li Zhimin Ou Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions Molecules alcohol dehydrogenase bienzyme cascade system chiral amines R-ω-transaminase recombinant engineered bacteria |
| title | Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions |
| title_full | Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions |
| title_fullStr | Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions |
| title_full_unstemmed | Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions |
| title_short | Efficient Synthesis of Key Chiral Intermediate in Painkillers (R)-1-[3,5-Bis(trifluoromethyl)phenyl]ethanamine by Bienzyme Cascade System with R-ω-Transaminase and Alcohol Dehydrogenase Functions |
| title_sort | efficient synthesis of key chiral intermediate in painkillers r 1 3 5 bis trifluoromethyl phenyl ethanamine by bienzyme cascade system with r ω transaminase and alcohol dehydrogenase functions |
| topic | alcohol dehydrogenase bienzyme cascade system chiral amines R-ω-transaminase recombinant engineered bacteria |
| url | https://www.mdpi.com/1420-3049/27/21/7331 |
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