한빛사논문
- 조회592
KAIST
So Young Choia,1, Jeonghun Ohb,1, JaeHwang Jungb,1,2, YongKeun Parkb,c,3, and Sang Yup Leea,d,3
aMetabolic and Biomolecular Engineering National Research Laboratory, Systems Metabolic Engineering and Systems Healthcare Cross-Generation Collaborative Laboratory, Department of Chemical and Biomolecular Engineering (BK21 four), Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea;
bDepartment of Physics, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea;
cTomocube Inc., Daejeon 34051, Republic of Korea;
dBioProcess Engineering Research Center and Bioinformatics Research Center, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea
↵1S.Y.C., J.O., and J.J. contributed equally to this work.
↵2Present address: Mechatronics R&D Center, Samsung Electronics, Hwasung-si 18448, Republic of Korea.
↵3To whom correspondence may be addressed.
Abstract
Polyhydroxyalkanoates (PHAs) are biodegradable polyesters that are intracellularly accumulated as distinct insoluble granules by various microorganisms. PHAs have attracted much attention as sustainable substitutes for petroleum-based plastics. However, the formation of PHA granules and their characteristics, such as localization, volume, weight, and density of granules, in an individual live bacterial cell are not well understood. Here, we report the results of three-dimensional (3D) quantitative label-free analysis of PHA granules in individual live bacterial cells through measuring the refractive index distributions by optical diffraction tomography (ODT). The formation and growth of PHA granules in the cells of Cupriavidus necator, the best-studied native PHA producer, and recombinant Escherichia coli harboring C. necator poly(3-hydroxybutyrate) (PHB) biosynthesis pathway are comparatively examined. Through the statistical ODT analyses of the bacterial cells, the distinctive characteristics for density and localization of PHB granules in vivo could be observed. The PHB granules in recombinant E. coli show higher density and localization polarity compared with those of C. necator, indicating that polymer chains are more densely packed and granules tend to be located at the cell poles, respectively. The cells were investigated in more detail through real-time 3D analyses, showing how differently PHA granules are processed in relation to the cell division process in native and nonnative PHA-producing strains. We also show that PHA granule–associated protein PhaM of C. necator plays a key role in making these differences between C. necator and recombinant E. coli strains. This study provides spatiotemporal insights into PHA accumulation inside the native and recombinant bacterial cells.
논문정보
- Research article
- 2021년 08월 (BRIC 등록일 2021-08-10)
- 국내 연구진
- 생명과학 > 응용생물화학
관련 웨비나
![홀로토모그래피를 이용한 살아있는 미생물 내 바이오 플라스틱 생성 정량 [Proc. Natl. Acad. Sci. USA]](/upload/board/files/onseminar/thumb/thumb_0000347.png)
학술웨비나
홀로토모그래피를 이용한 살아있는 미생물 내 바이오 플라스틱 생성 정량 [Proc. Natl. Acad. Sci. USA] 플라스틱 폐기물로 인한 환경오염 및 생태계 파괴는 전세계적인 문제가 되었고, 다양한 대안 기술들이 연구되고 있다. 그 중 미생물로부터 만들어지는 생분해성 폴리에스테르인 polyhydroxyalkanoate (PHA)가 기존 합성 플라스틱을 대체할 친환경 바이오 플라스틱으로 많은 관심을 받고 있다. 미생물을 이용하여 생산하는 PHA의 정량화는 연구 뿐만 아니라 바이오 플라스틱 생산 수율 관리에서도 중요한 주제이나, 크로마토그래피 등을 이용하는 기존 방식은 높은 농도의 미생물 시편에서 미생물을 모두 파괴한 후 측정이 가능했다. 본 연구에서는 홀로토모그래피 기술을 이용하여, 살아있는 미생물 내 PHA를 3차원 영상화와 정량화하고 또한 실시간 모니터링을 통해 약 8시간 동안 세포와 PHA과립 성장과정을 정밀하게 측정할 수 있음을 소개한다.- 박용근(KAIST)
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