Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions
Abstract Background Formate converted from CO2 reduction has great potential as a sustainable feedstock for biological production of biofuels and biochemicals. Nevertheless, utilization of formate for growth and chemical production by microbial species is limited due to its toxicity or the lack of a...
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| Format: | Article |
| Language: | English |
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BMC
2019-09-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-019-1547-z |
| _version_ | 1818228709484658688 |
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| author | Seung-Jin Kim Jihee Yoon Dae-Kyun Im Yong Hwan Kim Min-Kyu Oh |
| author_facet | Seung-Jin Kim Jihee Yoon Dae-Kyun Im Yong Hwan Kim Min-Kyu Oh |
| author_sort | Seung-Jin Kim |
| collection | DOAJ |
| description | Abstract Background Formate converted from CO2 reduction has great potential as a sustainable feedstock for biological production of biofuels and biochemicals. Nevertheless, utilization of formate for growth and chemical production by microbial species is limited due to its toxicity or the lack of a metabolic pathway. Here, we constructed a formate assimilation pathway in Escherichia coli and applied adaptive laboratory evolution to improve formate utilization as a carbon source in sugar-free conditions. Results The genes related to the tetrahydrofolate and serine cycles from Methylobacterium extorquens AM1 were overexpressed for formate assimilation, which was proved by the 13C-labeling experiments. The amino acids detected by GC/MS showed significant carbon labeling due to biomass production from formate. Then, 150 serial subcultures were performed to screen for evolved strains with improved ability to utilize formate. The genomes of evolved mutants were sequenced and the mutations were associated with formate dehydrogenation, folate metabolism, and biofilm formation. Last, 90 mg/L of ethanol production from formate was achieved using fed-batch cultivation without addition of sugars. Conclusion This work demonstrates the effectiveness of the introduction of a formate assimilation pathway, combined with adaptive laboratory evolution, to achieve the utilization of formate as a carbon source. This study suggests that the constructed E. coli could serve as a strain to exploit formate and captured CO2. |
| first_indexed | 2024-12-12T10:07:01Z |
| format | Article |
| id | doaj.art-079474a76e574031bf1ce8a07016fe0f |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T10:07:01Z |
| publishDate | 2019-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-079474a76e574031bf1ce8a07016fe0f2022-12-22T00:27:53ZengBMCBiotechnology for Biofuels1754-68342019-09-0112111210.1186/s13068-019-1547-zAdaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditionsSeung-Jin Kim0Jihee Yoon1Dae-Kyun Im2Yong Hwan Kim3Min-Kyu Oh4Department of Chemical and Biological Engineering, Korea UniversityDepartment of Chemical and Biological Engineering, Korea UniversityDepartment of Chemical and Biological Engineering, Korea UniversitySchool of Energy and Chemical Engineering, UNIST, Ulju-gunDepartment of Chemical and Biological Engineering, Korea UniversityAbstract Background Formate converted from CO2 reduction has great potential as a sustainable feedstock for biological production of biofuels and biochemicals. Nevertheless, utilization of formate for growth and chemical production by microbial species is limited due to its toxicity or the lack of a metabolic pathway. Here, we constructed a formate assimilation pathway in Escherichia coli and applied adaptive laboratory evolution to improve formate utilization as a carbon source in sugar-free conditions. Results The genes related to the tetrahydrofolate and serine cycles from Methylobacterium extorquens AM1 were overexpressed for formate assimilation, which was proved by the 13C-labeling experiments. The amino acids detected by GC/MS showed significant carbon labeling due to biomass production from formate. Then, 150 serial subcultures were performed to screen for evolved strains with improved ability to utilize formate. The genomes of evolved mutants were sequenced and the mutations were associated with formate dehydrogenation, folate metabolism, and biofilm formation. Last, 90 mg/L of ethanol production from formate was achieved using fed-batch cultivation without addition of sugars. Conclusion This work demonstrates the effectiveness of the introduction of a formate assimilation pathway, combined with adaptive laboratory evolution, to achieve the utilization of formate as a carbon source. This study suggests that the constructed E. coli could serve as a strain to exploit formate and captured CO2.http://link.springer.com/article/10.1186/s13068-019-1547-zFormateEscherichia coliAdaptive laboratory evolutionCarbon-labeling experiment |
| spellingShingle | Seung-Jin Kim Jihee Yoon Dae-Kyun Im Yong Hwan Kim Min-Kyu Oh Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions Biotechnology for Biofuels Formate Escherichia coli Adaptive laboratory evolution Carbon-labeling experiment |
| title | Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions |
| title_full | Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions |
| title_fullStr | Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions |
| title_full_unstemmed | Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions |
| title_short | Adaptively evolved Escherichia coli for improved ability of formate utilization as a carbon source in sugar-free conditions |
| title_sort | adaptively evolved escherichia coli for improved ability of formate utilization as a carbon source in sugar free conditions |
| topic | Formate Escherichia coli Adaptive laboratory evolution Carbon-labeling experiment |
| url | http://link.springer.com/article/10.1186/s13068-019-1547-z |
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