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김필
김필 (Pil Kim) 저자 이메일 보기
가톨릭대학교
 
조회 2073  인쇄하기 주소복사 트위터 공유 페이스북 공유 
An evolutionary optimization of a rhodopsin-based phototrophic metabolism in Escherichia coli
열기 Authors and Affiliations

Abstract

Background
The expression of the Gloeobacter rhodopsin (GR) in a chemotrophic Escherichia coli enables the light-driven phototrophic energy generation. Adaptive laboratory evolution has been used for acquiring desired phenotype of microbial cells and for the elucidation of basic mechanism of molecular evolution. To develop an optimized strain for the artificially acquired phototrophic metabolism, an ancestral E. coli expressing GR was adaptively evolved in a chemostat reactor with constant illumination and limited glucose conditions. This study was emphasized at an unexpected genomic mutation contributed to the improvement of microbial performance.

Results
During the chemostat culture, increase of cell size was observed, which were distinguished from that of the typical rod-shaped ancestral cells. A descendant ET5 strain was randomly isolated from the chemostat culture at 88-days. The phototrophic growth and the light-induced proton pumping of the ET5 strain were twofold and eightfold greater, respectively, than those of the ancestral E. coli strain. Single point mutation of C1082A at dgcQ gene (encoding diguanylate cyclase, also known as the yedQ gene) in the chromosome of ET5 strain was identified from whole genome sequencing analysis. An ancestral E. coli complemented with the same dgcQ mutation from the ET5 was repeated the subsequently enhancements of light-driven phototrophic growth and proton pumping. Intracellular c-di-GMP, the product of the diguanylate cyclase (dgcQ), of the descendant ET5 strain was suddenly increased while that of the ancestral strain was negligible.

Conclusions
Newly acquired phototrophic metabolism of E. coli was further improved via adaptive laboratory evolution by the rise of a point mutation on a transmembrane cell signaling protein followed by increase of signal molecule that eventually led an increase proton pumping and phototrophic growth.

Keywords : Adaptive laboratory evolution, Strain optimization, Chemotroph, Phototroph, Rhodopsin, Proton pumping

논문정보 F1000선정
- 형식: Research article
- 게재일: 2017년 06월 (BRIC 등록일 2017-07-14)
- 연구진: 국내연구진태극기
- 분야: Biotechnology, Microbiology
- 추천: Faculty of 1000 Biology
- 추천사유: F1000Prime Recommendations, Dissents and Comments for [Kim HA et al., Microb Cell Fact 2017, 16(1):111]. In F1000Prime, 17 Jul 2017; F1000Prime.com/727739764
CRISPR-BetaBinomial(CB2) 기법을 이용한 CRISPR pooled screen data 분석[Genome Res.]
정현환
발표: 정현환 (Baylor College of Medicine, Texas C...)
일자: 2019년 6월 26일 (수) 오전 10시 (한국시간)
언어:
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Springer Nature
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김현 (서울대학교)
김현주 (중앙대학교)
이상준 (중앙대학교)
관련분야 논문보기
Biotechnology

Microbiology

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