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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Autores principales: Hyang-Mi Lee, Jun Ren, Myeong-Sang Yu, Hyunjoo Kim, Woo Young Kim, Junhao Shen, Seung Min Yoo, Seong-il Eyun, Dokyun Na
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Publicado: BMC 2021
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Acceso en línea:https://doaj.org/article/483c0706133f4ef6885765b9040a9b44
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spelling oai:doaj.org-article:483c0706133f4ef6885765b9040a9b442021-12-05T12:16:43ZConstruction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production10.1186/s13068-021-02077-81754-6834https://doaj.org/article/483c0706133f4ef6885765b9040a9b442021-12-01T00:00:00Zhttps://doi.org/10.1186/s13068-021-02077-8https://doaj.org/toc/1754-6834Abstract 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 AbstractHyang-Mi LeeJun RenMyeong-Sang YuHyunjoo KimWoo Young KimJunhao ShenSeung Min YooSeong-il EyunDokyun NaBMCarticleMethylomonas sp. DH-1Promoter libraryGene expression optimizationCadaverineFuelTP315-360BiotechnologyTP248.13-248.65ENBiotechnology for Biofuels, Vol 14, Iss 1, Pp 1-12 (2021)
institution DOAJ
collection DOAJ
language EN
topic Methylomonas sp. DH-1
Promoter library
Gene expression optimization
Cadaverine
Fuel
TP315-360
Biotechnology
TP248.13-248.65
spellingShingle Methylomonas sp. DH-1
Promoter library
Gene expression optimization
Cadaverine
Fuel
TP315-360
Biotechnology
TP248.13-248.65
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
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
format article
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
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_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_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_sort construction of a tunable promoter library to optimize gene expression in methylomonas sp. dh-1, a methanotroph, and its application to cadaverine production
publisher BMC
publishDate 2021
url https://doaj.org/article/483c0706133f4ef6885765b9040a9b44
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