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- Title
Construction of a tunable promoter library to optimize gene expression in Methylomonas sp. DH-1, a methanotroph, and its application to cadaverine production.
- Authors
Lee, Hyang-Mi; Ren, Jun; Yu, Myeong-Sang; Kim, Hyunjoo; Kim, Woo Young; Shen, Junhao; Yoo, Seung Min; Eyun, Seong-il; Na, Dokyun
- 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.
- Subjects
GENE libraries; METHANOTROPHS; GREENHOUSE effect; GENETIC engineering; CARBON dioxide; GENE expression
- Publication
Biotechnology for Biofuels, 2021, Vol 14, Issue 1, p1
- ISSN
1754-6834
- Publication type
Article
- DOI
10.1186/s13068-021-02077-8