Genome Variations of Evolved Escherichia coli ET8 With a Rhodopsin-Based Phototrophic Metabolism
- Authors
- Kim, Hyun Aaron; Kim, Hyun Ju; Lee, Min Ju; Park, Jihoon; Choi, Ah Reum; Jeong, Haeyoung; Jung, Kwang-Hwan; Kim, Pil; Lee, Sang Jun
- Issue Date
- Jul-2018
- Publisher
- WILEY-V C H VERLAG GMBH
- Keywords
- adaptive laboratory evolution; chemotroph; kdtA-upregulation; galU-disruption; Gloeobacter rhodopsin; phototroph
- Citation
- BIOTECHNOLOGY JOURNAL, v.13, no.7
- Journal Title
- BIOTECHNOLOGY JOURNAL
- Volume
- 13
- Number
- 7
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/2012
- DOI
- 10.1002/biot.201700497
- ISSN
- 1860-6768
1860-7314
- Abstract
- We reported that the phototrophic metabolism via plasmid-originated Gloeobacter rhodopsin(GR)-expression is improved in Escherichia coli ET5 harboring pKJ606-GR by a genomic point mutation (dgcQ(C1082A)) encoding a transmembrane cell signaling protein (Microb. Cell Fact. 16:111, 2017). Another evolved descendant is isolated from the chemostat, and the genome variation of the strain named ET8 harboring pKJ606-GR is investigated in this study. Whole genome sequencing analysis identifies a single point mutation (C3831976A) located in the non-coding upstream region of kdtA and an IS4 insertional mutation at galU(G706) without any mutations in the plasmid. ET8 strain shows enhanced kdtA transcription and no growth in the D-galactose or lactose sole carbon sourced minimal media. Size of ET8 strain are almost identical to that of the ancestor. Phototrophic growth and proton pumping in ET8 expressing GR (ET8+GR) are increased 1.5-fold and threefold, respectively, compared with those in the ancestor (W3110+GR). To verify the effects of the genomic mutations, either the kdtA-upregulation or the galU-disruption is conducted in the ancestor. Both the kdtA-upregulation and the galU-disruption result in the drastic increases of proton-pumping. The physiological properties arising from the genomic variations of the evolved host with the new phototrophic metabolism are further discussed.
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