Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid
The U.S. Department of Energy has listed levulinic acid (LA) as one of the top 12 compounds derived from biomass. LA has gained much attention owing to its conversion into enantiopure 4-aminopentanoic acid through an amination reaction. Herein, we developed a coupled-enzyme recyclable cascade employ...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-11-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1280464/full |
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| author | Taresh P. Khobragade Pritam Giri Amol D. Pagar Mahesh D. Patil Sharad Sarak Sangwoo Joo Younghwan Goh Seohee Jung Hyunseok Yoon Subin Yun Youkyoung Kwon Hyungdon Yun |
| author_facet | Taresh P. Khobragade Pritam Giri Amol D. Pagar Mahesh D. Patil Sharad Sarak Sangwoo Joo Younghwan Goh Seohee Jung Hyunseok Yoon Subin Yun Youkyoung Kwon Hyungdon Yun |
| author_sort | Taresh P. Khobragade |
| collection | DOAJ |
| description | The U.S. Department of Energy has listed levulinic acid (LA) as one of the top 12 compounds derived from biomass. LA has gained much attention owing to its conversion into enantiopure 4-aminopentanoic acid through an amination reaction. Herein, we developed a coupled-enzyme recyclable cascade employing two transaminases (TAs) for the synthesis of (S)-4-aminopentanoic acid. TAs were first utilized to convert LA into (S)-4-aminopentanoic acid using (S)-α-Methylbenzylamine [(S)-α-MBA] as an amino donor. The deaminated (S)-α-MBA i.e., acetophenone was recycled back using a second TAs while using isopropyl amine (IPA) amino donor to generate easily removable acetone. Enzymatic reactions were carried out using different systems, with conversions ranging from 30% to 80%. Furthermore, the hybrid nanoflowers (HNF) of the fusion protein were constructed which afforded complete biocatalytic conversion of LA to the desired (S)-4-aminopentanoic acid. The created HNF demonstrated storage stability for over a month and can be reused for up to 7 sequential cycles. A preparative scale reaction (100 mL) achieved the complete conversion with an isolated yield of 62%. Furthermore, the applicability of this recycling system was tested with different β-keto ester substrates, wherein 18%–48% of corresponding β-amino acids were synthesized. Finally, this recycling system was applied for the biosynthesis of pharmaceutical important drug sitagliptin intermediate ((R)-3-amino-4-(2,4,5-triflurophenyl) butanoic acid) with an excellent conversion 82%. |
| first_indexed | 2024-03-11T09:26:34Z |
| format | Article |
| id | doaj.art-d57c69284d754bcd9af24bac24b15c2a |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-03-11T09:26:34Z |
| publishDate | 2023-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-d57c69284d754bcd9af24bac24b15c2a2023-11-16T17:59:07ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-11-011110.3389/fbioe.2023.12804641280464Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acidTaresh P. Khobragade0Pritam Giri1Amol D. Pagar2Mahesh D. Patil3Sharad Sarak4Sangwoo Joo5Younghwan Goh6Seohee Jung7Hyunseok Yoon8Subin Yun9Youkyoung Kwon10Hyungdon Yun11Department of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Nanomaterials and Application Technology, Center of Innovative and Applied Bioprocessing (CIAB), Mohali, Punjab, IndiaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaDepartment of Systems Biotechnology, Konkuk University, Seoul, Republic of KoreaThe U.S. Department of Energy has listed levulinic acid (LA) as one of the top 12 compounds derived from biomass. LA has gained much attention owing to its conversion into enantiopure 4-aminopentanoic acid through an amination reaction. Herein, we developed a coupled-enzyme recyclable cascade employing two transaminases (TAs) for the synthesis of (S)-4-aminopentanoic acid. TAs were first utilized to convert LA into (S)-4-aminopentanoic acid using (S)-α-Methylbenzylamine [(S)-α-MBA] as an amino donor. The deaminated (S)-α-MBA i.e., acetophenone was recycled back using a second TAs while using isopropyl amine (IPA) amino donor to generate easily removable acetone. Enzymatic reactions were carried out using different systems, with conversions ranging from 30% to 80%. Furthermore, the hybrid nanoflowers (HNF) of the fusion protein were constructed which afforded complete biocatalytic conversion of LA to the desired (S)-4-aminopentanoic acid. The created HNF demonstrated storage stability for over a month and can be reused for up to 7 sequential cycles. A preparative scale reaction (100 mL) achieved the complete conversion with an isolated yield of 62%. Furthermore, the applicability of this recycling system was tested with different β-keto ester substrates, wherein 18%–48% of corresponding β-amino acids were synthesized. Finally, this recycling system was applied for the biosynthesis of pharmaceutical important drug sitagliptin intermediate ((R)-3-amino-4-(2,4,5-triflurophenyl) butanoic acid) with an excellent conversion 82%.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1280464/fullbiomassamino donortransaminasenanoflowerfusion protein |
| spellingShingle | Taresh P. Khobragade Pritam Giri Amol D. Pagar Mahesh D. Patil Sharad Sarak Sangwoo Joo Younghwan Goh Seohee Jung Hyunseok Yoon Subin Yun Youkyoung Kwon Hyungdon Yun Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid Frontiers in Bioengineering and Biotechnology biomass amino donor transaminase nanoflower fusion protein |
| title | Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid |
| title_full | Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid |
| title_fullStr | Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid |
| title_full_unstemmed | Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid |
| title_short | Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid |
| title_sort | dual function transaminases with hybrid nanoflower for the production of value added chemicals from biobased levulinic acid |
| topic | biomass amino donor transaminase nanoflower fusion protein |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1280464/full |
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