Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials
Acetoin is an important four-carbon platform chemical with versatile applications. Optically pure (<i>R</i>)-acetoin is more valuable than the racemate as it can be applied in the asymmetric synthesis of optically active α-hydroxy ketone derivatives, pharmaceuticals, and liquid crystal c...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/1/203 |
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| author | Mengxue Diao Xianrui Chen Jing Li Ya’nan Shi Bo Yu Zhilin Ma Jianxiu Li Nengzhong Xie |
| author_facet | Mengxue Diao Xianrui Chen Jing Li Ya’nan Shi Bo Yu Zhilin Ma Jianxiu Li Nengzhong Xie |
| author_sort | Mengxue Diao |
| collection | DOAJ |
| description | Acetoin is an important four-carbon platform chemical with versatile applications. Optically pure (<i>R</i>)-acetoin is more valuable than the racemate as it can be applied in the asymmetric synthesis of optically active α-hydroxy ketone derivatives, pharmaceuticals, and liquid crystal composites. As a cytotoxic solvent, acetoin at high concentrations severely limits culture performance and impedes the acetoin yield of cell factories. In this study, putative genes that may improve the resistance to acetoin for <i>Escherichia coli</i> were screened. To obtain a high-producing strain, the identified acetoin-resistance gene was overexpressed, and the synthetic pathway of (<i>R</i>)-acetoin was strengthened by optimizing the copy number of the key genes. The engineered <i>E</i>. <i>coli</i> strain GXASR-49RSF produced 81.62 g/L (<i>R</i>)-acetoin with an enantiomeric purity of 96.5% in the fed-batch fermentation using non-food raw materials in a 3-L fermenter. Combining the systematic approach developed in this study with the use of low-cost feedstock showed great potential for (<i>R</i>)-acetoin production via this cost-effective biotechnological process. |
| first_indexed | 2024-03-09T11:38:34Z |
| format | Article |
| id | doaj.art-c5b0946891464978a7a081402d74eca0 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T11:38:34Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-c5b0946891464978a7a081402d74eca02023-11-30T23:37:55ZengMDPI AGMicroorganisms2076-26072023-01-0111120310.3390/microorganisms11010203Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw MaterialsMengxue Diao0Xianrui Chen1Jing Li2Ya’nan Shi3Bo Yu4Zhilin Ma5Jianxiu Li6Nengzhong Xie7State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-Food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, 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 Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Life Science and Technology College, Guangxi University, Nanning 530004, ChinaCAS Key Laboratory of Microbial Physiological & Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, ChinaCAS Key Laboratory of Microbial Physiological & Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Life Science and Technology College, Guangxi University, Nanning 530004, 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 Academy of Sciences, 98 Daling Road, Nanning 530007, 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 Academy of Sciences, 98 Daling Road, Nanning 530007, ChinaAcetoin is an important four-carbon platform chemical with versatile applications. Optically pure (<i>R</i>)-acetoin is more valuable than the racemate as it can be applied in the asymmetric synthesis of optically active α-hydroxy ketone derivatives, pharmaceuticals, and liquid crystal composites. As a cytotoxic solvent, acetoin at high concentrations severely limits culture performance and impedes the acetoin yield of cell factories. In this study, putative genes that may improve the resistance to acetoin for <i>Escherichia coli</i> were screened. To obtain a high-producing strain, the identified acetoin-resistance gene was overexpressed, and the synthetic pathway of (<i>R</i>)-acetoin was strengthened by optimizing the copy number of the key genes. The engineered <i>E</i>. <i>coli</i> strain GXASR-49RSF produced 81.62 g/L (<i>R</i>)-acetoin with an enantiomeric purity of 96.5% in the fed-batch fermentation using non-food raw materials in a 3-L fermenter. Combining the systematic approach developed in this study with the use of low-cost feedstock showed great potential for (<i>R</i>)-acetoin production via this cost-effective biotechnological process.https://www.mdpi.com/2076-2607/11/1/203(<i>R</i>)-acetoinacetoin-tolerancesynthetic biologylow-cost feedstock |
| spellingShingle | Mengxue Diao Xianrui Chen Jing Li Ya’nan Shi Bo Yu Zhilin Ma Jianxiu Li Nengzhong Xie Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials Microorganisms (<i>R</i>)-acetoin acetoin-tolerance synthetic biology low-cost feedstock |
| title | Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials |
| title_full | Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials |
| title_fullStr | Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials |
| title_full_unstemmed | Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials |
| title_short | Metabolic Engineering of <i>Escherichia coli</i> for High-Level Production of (<i>R</i>)-Acetoin from Low-Cost Raw Materials |
| title_sort | metabolic engineering of i escherichia coli i for high level production of i r i acetoin from low cost raw materials |
| topic | (<i>R</i>)-acetoin acetoin-tolerance synthetic biology low-cost feedstock |
| url | https://www.mdpi.com/2076-2607/11/1/203 |
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