High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli
Biosynthesis of the functional factor γ-aminobutyric acid (GABA) in bacteria involves two key proteins an intracellular glutamate decarboxylase (GadB) and a membrane-bound antiporter (GadC). Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity. In t...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | Engineering Microbiology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667370323000097 |
| _version_ | 1797788377863421952 |
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| author | Lili Yao Changjiang Lyu Yuting Wang Sheng Hu Weirui Zhao Hongwei Cao Jun Huang Lehe Mei |
| author_facet | Lili Yao Changjiang Lyu Yuting Wang Sheng Hu Weirui Zhao Hongwei Cao Jun Huang Lehe Mei |
| author_sort | Lili Yao |
| collection | DOAJ |
| description | Biosynthesis of the functional factor γ-aminobutyric acid (GABA) in bacteria involves two key proteins an intracellular glutamate decarboxylase (GadB) and a membrane-bound antiporter (GadC). Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity. In this study, gadBΔC11 of Lactiplantibacillus plantarum and gadCΔC41 of Escherichia coli were inserted into the designed double promoter (PT7lac and PBAD) expression system. Then, E. coli Lemo21(DE3) was chosen as the host to minimize the toxic effects of GadCΔC41 overexpression. Furthermore, a green and high-efficiency GABA synthesis system using dormant engineered Lemo21(DE3) cells as biocatalysts was developed. The total GABA yield reached 829.08 g/L with a 98.7% conversion ratio within 13 h, when engineered E. coli Lemo21(DE3) cells were concentrated to an OD600 of 20 and reused for three cycles in a 3 M L-glutamate solution at 37 °C, which represented the highest GABA productivity ever reported. Overall, expanding the active pH ranges of GadB and GadC toward physiological pH and employing a tunable expression host for membrane-bound GadC production is a promising strategy for high-level GABA biosynthesis in E. coli. |
| first_indexed | 2024-03-13T01:34:41Z |
| format | Article |
| id | doaj.art-39f575b4ed844185b53710308b3dbc7c |
| institution | Directory Open Access Journal |
| issn | 2667-3703 |
| language | English |
| last_indexed | 2024-03-13T01:34:41Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Engineering Microbiology |
| spelling | doaj.art-39f575b4ed844185b53710308b3dbc7c2023-07-04T05:11:29ZengElsevierEngineering Microbiology2667-37032023-06-0132100077High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coliLili Yao0Changjiang Lyu1Yuting Wang2Sheng Hu3Weirui Zhao4Hongwei Cao5Jun Huang6Lehe Mei7College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China; School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, ChinaSchool of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China; Corresponding authors.School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, ChinaSchool of Biological and Chemical Engineering, Ningbo Tech University, Ningbo 315100, ChinaSchool of Biological and Chemical Engineering, Ningbo Tech University, Ningbo 315100, ChinaCollege of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China; Corresponding authors.School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China; Corresponding authors.School of Biological and Chemical Engineering, Ningbo Tech University, Ningbo 315100, China; College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, ChinaBiosynthesis of the functional factor γ-aminobutyric acid (GABA) in bacteria involves two key proteins an intracellular glutamate decarboxylase (GadB) and a membrane-bound antiporter (GadC). Efficient co-expression of suitable GadB and GadC candidates is crucial for improving GABA productivity. In this study, gadBΔC11 of Lactiplantibacillus plantarum and gadCΔC41 of Escherichia coli were inserted into the designed double promoter (PT7lac and PBAD) expression system. Then, E. coli Lemo21(DE3) was chosen as the host to minimize the toxic effects of GadCΔC41 overexpression. Furthermore, a green and high-efficiency GABA synthesis system using dormant engineered Lemo21(DE3) cells as biocatalysts was developed. The total GABA yield reached 829.08 g/L with a 98.7% conversion ratio within 13 h, when engineered E. coli Lemo21(DE3) cells were concentrated to an OD600 of 20 and reused for three cycles in a 3 M L-glutamate solution at 37 °C, which represented the highest GABA productivity ever reported. Overall, expanding the active pH ranges of GadB and GadC toward physiological pH and employing a tunable expression host for membrane-bound GadC production is a promising strategy for high-level GABA biosynthesis in E. coli.http://www.sciencedirect.com/science/article/pii/S2667370323000097Glutamate decarboxylaseAntiporterγ-Aminobutyric acidE. coliWhole-cell biocatalysts |
| spellingShingle | Lili Yao Changjiang Lyu Yuting Wang Sheng Hu Weirui Zhao Hongwei Cao Jun Huang Lehe Mei High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli Engineering Microbiology Glutamate decarboxylase Antiporter γ-Aminobutyric acid E. coli Whole-cell biocatalysts |
| title | High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli |
| title_full | High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli |
| title_fullStr | High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli |
| title_full_unstemmed | High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli |
| title_short | High-level production of γ-aminobutyric acid via efficient co-expression of the key genes of glutamate decarboxylase system in Escherichia coli |
| title_sort | high level production of γ aminobutyric acid via efficient co expression of the key genes of glutamate decarboxylase system in escherichia coli |
| topic | Glutamate decarboxylase Antiporter γ-Aminobutyric acid E. coli Whole-cell biocatalysts |
| url | http://www.sciencedirect.com/science/article/pii/S2667370323000097 |
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