A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine
Abstract In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d-p-hydroxyphenylglycine (D-HPG), a valuable intermediate used to produce β-lactam antibiotics and in fine-chemical synthesis, from l-tyrosine. In this pathway, catalytic conversion of...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2021-05-01
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| Series: | Bioresources and Bioprocessing |
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| Online Access: | https://doi.org/10.1186/s40643-021-00394-2 |
| _version_ | 1819081821884776448 |
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| author | Xu Tan Sheng Zhang Wei Song Jia Liu Cong Gao Xiulai Chen Liming Liu Jing Wu |
| author_facet | Xu Tan Sheng Zhang Wei Song Jia Liu Cong Gao Xiulai Chen Liming Liu Jing Wu |
| author_sort | Xu Tan |
| collection | DOAJ |
| description | Abstract In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d-p-hydroxyphenylglycine (D-HPG), a valuable intermediate used to produce β-lactam antibiotics and in fine-chemical synthesis, from l-tyrosine. In this pathway, catalytic conversion of the intermediate 4-hydroxyphenylglyoxalate by meso-diaminopimelate dehydrogenase from Corynebacterium glutamicum (CgDAPDH) was identified as the rate-limiting step, followed by application of a mechanism-guided “conformation rotation” strategy to decrease the hydride-transfer distance d(C6HDAP−C4NNADP) and increase CgDAPDH activity. Introduction of the best variant generated by protein engineering (CgDAPDHBC621/D120S/W144S/I169P with 5.32 ± 0.85 U·mg−1 specific activity) into the designed pathway resulted in a D-HPG titer of 42.69 g/L from 50-g/L l-tyrosine in 24 h, with 92.5% conversion, 71.5% isolated yield, and > 99% enantiomeric excess in a 3-L fermenter. This four-enzyme cascade provides an efficient enzymatic approach for the industrial production of D-HPG from cheap amino acids. |
| first_indexed | 2024-12-21T20:06:52Z |
| format | Article |
| id | doaj.art-ace5a9df5f474b5985c962258d0e855e |
| institution | Directory Open Access Journal |
| issn | 2197-4365 |
| language | English |
| last_indexed | 2024-12-21T20:06:52Z |
| publishDate | 2021-05-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Bioresources and Bioprocessing |
| spelling | doaj.art-ace5a9df5f474b5985c962258d0e855e2022-12-21T18:51:49ZengSpringerOpenBioresources and Bioprocessing2197-43652021-05-018111510.1186/s40643-021-00394-2A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosineXu Tan0Sheng Zhang1Wei Song2Jia Liu3Cong Gao4Xiulai Chen5Liming Liu6Jing Wu7School of Pharmaceutical Science, Jiangnan UniversityZhejiang Tianrui Chemical Co., LtdSchool of Pharmaceutical Science, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversityState Key Laboratory of Food Science and Technology, Jiangnan UniversitySchool of Pharmaceutical Science, Jiangnan UniversityAbstract In this study, a four-enzyme cascade pathway was developed and reconstructed in vivo for the production of d-p-hydroxyphenylglycine (D-HPG), a valuable intermediate used to produce β-lactam antibiotics and in fine-chemical synthesis, from l-tyrosine. In this pathway, catalytic conversion of the intermediate 4-hydroxyphenylglyoxalate by meso-diaminopimelate dehydrogenase from Corynebacterium glutamicum (CgDAPDH) was identified as the rate-limiting step, followed by application of a mechanism-guided “conformation rotation” strategy to decrease the hydride-transfer distance d(C6HDAP−C4NNADP) and increase CgDAPDH activity. Introduction of the best variant generated by protein engineering (CgDAPDHBC621/D120S/W144S/I169P with 5.32 ± 0.85 U·mg−1 specific activity) into the designed pathway resulted in a D-HPG titer of 42.69 g/L from 50-g/L l-tyrosine in 24 h, with 92.5% conversion, 71.5% isolated yield, and > 99% enantiomeric excess in a 3-L fermenter. This four-enzyme cascade provides an efficient enzymatic approach for the industrial production of D-HPG from cheap amino acids.https://doi.org/10.1186/s40643-021-00394-2d-p-hydroxyphenylglycinemeso-diaminopimelate dehydrogenaseHydride transfer distancel-tyrosineProtein engineering |
| spellingShingle | Xu Tan Sheng Zhang Wei Song Jia Liu Cong Gao Xiulai Chen Liming Liu Jing Wu A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine Bioresources and Bioprocessing d-p-hydroxyphenylglycine meso-diaminopimelate dehydrogenase Hydride transfer distance l-tyrosine Protein engineering |
| title | A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine |
| title_full | A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine |
| title_fullStr | A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine |
| title_full_unstemmed | A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine |
| title_short | A multi-enzyme cascade for efficient production of d-p-hydroxyphenylglycine from l-tyrosine |
| title_sort | multi enzyme cascade for efficient production of d p hydroxyphenylglycine from l tyrosine |
| topic | d-p-hydroxyphenylglycine meso-diaminopimelate dehydrogenase Hydride transfer distance l-tyrosine Protein engineering |
| url | https://doi.org/10.1186/s40643-021-00394-2 |
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