Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production
Abstract Background As methane is 84 times more potent than carbon dioxide in exacerbating the greenhouse effect, there is an increasing interest in the utilization of methanotrophic bacteria that can convert harmful methane into various value-added compounds. A recently isolated methanotroph, Methy...
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| Format: | Article |
| Language: | English |
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BMC
2021-12-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-021-02077-8 |
| _version_ | 1828724145486036992 |
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| author | Hyang-Mi Lee Jun Ren Myeong-Sang Yu Hyunjoo Kim Woo Young Kim Junhao Shen Seung Min Yoo Seong-il Eyun Dokyun Na |
| author_facet | Hyang-Mi Lee Jun Ren Myeong-Sang Yu Hyunjoo Kim Woo Young Kim Junhao Shen Seung Min Yoo Seong-il Eyun Dokyun Na |
| author_sort | Hyang-Mi Lee |
| collection | DOAJ |
| description | Abstract Background As methane is 84 times more potent than carbon dioxide in exacerbating the greenhouse effect, there is an increasing interest in the utilization of methanotrophic bacteria that can convert harmful methane into various value-added compounds. A recently isolated methanotroph, Methylomonas sp. DH-1, is a promising biofactory platform because of its relatively fast growth. However, the lack of genetic engineering tools hampers its wide use in the bioindustry. Results Through three different approaches, we constructed a tunable promoter library comprising 33 promoters that can be used for the metabolic engineering of Methylomonas sp. DH-1. The library had an expression level of 0.24–410% when compared with the strength of the lac promoter. For practical application of the promoter library, we fine-tuned the expressions of cadA and cadB genes, required for cadaverine synthesis and export, respectively. The strain with PrpmB-cadA and PDnaA-cadB produced the highest cadaverine titre (18.12 ± 1.06 mg/L) in Methylomonas sp. DH-1, which was up to 2.8-fold higher than that obtained from a non-optimized strain. In addition, cell growth and lysine (a precursor of cadaverine) production assays suggested that gene expression optimization through transcription tuning can afford a balance between the growth and precursor supply. Conclusions The tunable promoter library provides standard and tunable components for gene expression, thereby facilitating the use of methanotrophs, specifically Methylomonas sp. DH-1, as a sustainable cell factory. Graphical Abstract |
| first_indexed | 2024-04-12T13:03:30Z |
| format | Article |
| id | doaj.art-483c0706133f4ef6885765b9040a9b44 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-12T13:03:30Z |
| publishDate | 2021-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-483c0706133f4ef6885765b9040a9b442022-12-22T03:32:06ZengBMCBiotechnology for Biofuels1754-68342021-12-0114111210.1186/s13068-021-02077-8Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine productionHyang-Mi Lee0Jun Ren1Myeong-Sang Yu2Hyunjoo Kim3Woo Young Kim4Junhao Shen5Seung Min Yoo6Seong-il Eyun7Dokyun Na8Department of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityDepartment of Life Science, Chung-Ang UniversityDepartment of Biomedical Engineering, Chung-Ang UniversityAbstract Background As methane is 84 times more potent than carbon dioxide in exacerbating the greenhouse effect, there is an increasing interest in the utilization of methanotrophic bacteria that can convert harmful methane into various value-added compounds. A recently isolated methanotroph, Methylomonas sp. DH-1, is a promising biofactory platform because of its relatively fast growth. However, the lack of genetic engineering tools hampers its wide use in the bioindustry. Results Through three different approaches, we constructed a tunable promoter library comprising 33 promoters that can be used for the metabolic engineering of Methylomonas sp. DH-1. The library had an expression level of 0.24–410% when compared with the strength of the lac promoter. For practical application of the promoter library, we fine-tuned the expressions of cadA and cadB genes, required for cadaverine synthesis and export, respectively. The strain with PrpmB-cadA and PDnaA-cadB produced the highest cadaverine titre (18.12 ± 1.06 mg/L) in Methylomonas sp. DH-1, which was up to 2.8-fold higher than that obtained from a non-optimized strain. In addition, cell growth and lysine (a precursor of cadaverine) production assays suggested that gene expression optimization through transcription tuning can afford a balance between the growth and precursor supply. Conclusions The tunable promoter library provides standard and tunable components for gene expression, thereby facilitating the use of methanotrophs, specifically Methylomonas sp. DH-1, as a sustainable cell factory. Graphical Abstracthttps://doi.org/10.1186/s13068-021-02077-8Methylomonas sp. DH-1Promoter libraryGene expression optimizationCadaverine |
| spellingShingle | Hyang-Mi Lee Jun Ren Myeong-Sang Yu Hyunjoo Kim Woo Young Kim Junhao Shen Seung Min Yoo Seong-il Eyun Dokyun Na Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production Biotechnology for Biofuels Methylomonas sp. DH-1 Promoter library Gene expression optimization Cadaverine |
| title | Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production |
| title_full | Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production |
| title_fullStr | Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production |
| title_full_unstemmed | Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production |
| title_short | Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production |
| title_sort | construction of a tunable promoter library to optimize gene expression in methylomonas sp dh 1 a methanotroph and its application to cadaverine production |
| topic | Methylomonas sp. DH-1 Promoter library Gene expression optimization Cadaverine |
| url | https://doi.org/10.1186/s13068-021-02077-8 |
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