[논문]Three-dimensional label-free visualization and quantification of polyhydroxyalkanoates in individual bacterial cell in its native state

Choi, So Young; Oh, Jeonghun; Jung, JaeHwang; Park, YongKeun; Lee, Sang Yup

doi:10.1073/pnas.2103956118

[해외논문] Three-dimensional label-free visualization and quantification of polyhydroxyalkanoates in individual bacterial cell in its native state 원문보기

Proceedings of the National Academy of Sciences of the United States of America, v.118 no.31, 2021년, pp.e2103956118 - e2103956118

Choi, So Young (Metabolic 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) , Oh, Jeonghun , Jung, JaeHwang (Department of Physics, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea) , Park, YongKeun , Lee, Sang Yup (Department of Physics, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea)

Abstract ▼ AI-Helper

SignificancePolyhydroxyalkanoates (PHAs) are bacterial polyesters considered as sustainable and environmentally friendly substitutes for petroleum-based plastics. Despite great advances in PHA research, spatiotemporal characteristics of PHA granule formation and distribution in a live bacterial cell are not well understood. Here, we report the results of a three-dimensional (3D) analysis of live bacterial cells accumulating poly(3-hydroxybutyrate) (PHB), the best-studied member of PHAs. Optical diffraction tomography was applied for visualizing individual cells and in vivo PHB granules and quantifying their physical properties such as weight, volume, density, and localization by measuring 3D refractive index distributions. Comparative analyses of native and recombinant PHB-producing strains revealed distinctive characteristics of PHB accumulation, providing insights into spatiotemporal localization of PHA granules in vivo.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.

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참고문헌 (26)

Anderson, A J, Dawes, E A. Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiological reviews, vol.54, no.4, 450-472.

상세보기
10.1002/(SICI)1097-0290(19960105)49:1<1::AID-BIT1>3.0.CO;2-P

상세보기
Chen, Guo-Qiang. A microbial polyhydroxyalkanoates (PHA) based bio- and materials industry. Chemical Society reviews, vol.38, no.8, 2434-2446.

상세보기
Choi, So Young, Rhie, Mi Na, Kim, Hee Taek, Joo, Jeong Chan, Cho, In Jin, Son, Jina, Jo, Seo Young, Sohn, Yu Jung, Baritugo, Kei-Anne, Pyo, Jiwon, Lee, Youngjoon, Lee, Sang Yup, Park, Si Jae. Metabolic engineering for the synthesis of polyesters: A 100-year journey from polyhydroxyalkanoates to non-natural microbial polyesters. Metabolic engineering, vol.58, 47-81.

상세보기
Jendrossek, Dieter. Polyhydroxyalkanoate Granules Are Complex Subcellular Organelles (Carbonosomes). Journal of bacteriology, vol.191, no.10, 3195-3202.

상세보기
Jendrossek, Dieter, Pfeiffer, Daniel. New insights in the formation of polyhydroxyalkanoate granules (carbonosomes) and novel functions of poly(3‐hydroxybutyrate). Environmental microbiology, vol.16, no.8, 2357-2373.

상세보기
Peters, Verena, Rehm, Bernd H.A.. In vivo monitoring of PHA granule formation using GFP-labeled PHA synthases. FEMS microbiology letters, vol.248, no.1, 93-100.

상세보기
Mravec, Filip, Obruca, Stanislav, Krzyzanek, Vladislav, Sedlacek, Petr, Hrubanova, Kamila, Samek, Ota, Kucera, Dan, Benesova, Pavla, Nebesarova, Jana, Steinbüchel, Alexander. Accumulation of PHA granules inCupriavidus necatoras seen by confocal fluorescence microscopy. FEMS microbiology letters, vol.363, no.10, fnw094-.

상세보기
Jendrossek, D.. Fluorescence Microscopical Investigation of Poly(3-hydroxybutyrate) Granule Formation in Bacteria. Biomacromolecules, vol.6, no.2, 598-603.

상세보기
Tian, Jiamin, Sinskey, Anthony J., Stubbe, JoAnne. Kinetic Studies of Polyhydroxybutyrate Granule Formation in Wautersia eutropha H16 by Transmission Electron Microscopy. Journal of bacteriology, vol.187, no.11, 3814-3824.

상세보기
Wahl, Andreas, Schuth, Nora, Pfeiffer, Daniel, Nussberger, Stephan, Jendrossek, Dieter. PHB granules are attached to the nucleoid via PhaM in Ralstonia eutropha. BMC microbiology, vol.12, 262-262.

상세보기
Beeby, M., Cho, M., Stubbe, J., Jensen, G. J.. Growth and Localization of Polyhydroxybutyrate Granules in Ralstonia eutropha. Journal of bacteriology, vol.194, no.5, 1092-1099.

상세보기
Gerngross, T U, Reilly, P, Stubbe, J, Sinskey, A J, Peoples, O P. Immunocytochemical analysis of poly-beta-hydroxybutyrate (PHB) synthase in Alcaligenes eutrophus H16: localization of the synthase enzyme at the surface of PHB granules. Journal of bacteriology, vol.175, no.16, 5289-5293.

상세보기
10.1002/(SICI)1521-3927(20000201)21:2<77::AID-MARC77>3.0.CO;2-6

상세보기
Obruca, Stanislav, Sedlacek, Petr, Slaninova, Eva, Fritz, Ines, Daffert, Christina, Meixner, Katharina, Sedrlova, Zuzana, Koller, Martin. Novel unexpected functions of PHA granules. Applied microbiology and biotechnology, vol.104, no.11, 4795-4810.

상세보기
Stubbe, JoAnne, Tian, Jiamin. Polyhydroxyalkanoate (PHA) homeostasis: the role of the PHA synthase. Natural product reports : a journal of current developments in bio-organic chemistry, vol.20, no.5, 445-457.

상세보기
Jendrossek, Dieter, Selchow, Olaf, Hoppert, Michael. Poly(3-Hydroxybutyrate) Granules at the Early Stages of Formation Are Localized Close to the Cytoplasmic Membrane in Caryophanon latum. Applied and environmental microbiology, vol.73, no.2, 586-593.

상세보기
Bresan, Stephanie, Jendrossek, Dieter. New Insights into PhaM-PhaC-Mediated Localization of Polyhydroxybutyrate Granules in Ralstonia eutropha H16. Applied and environmental microbiology, vol.83, no.12, e00505-17-e00505-17.

상세보기
Pötter, Markus, Müller, Helena, Steinbüchel, Alexander. Influence of homologous phasins (PhaP) on PHA accumulation and regulation of their expression by the transcriptional repressor PhaR in Ralstonia eutropha H16. Microbiology, vol.151, no.3, 825-833.

상세보기
Neumann, Liv, Spinozzi, Francesco, Sinibaldi, Raffaele, Rustichelli, Franco, Pötter, Markus, Steinbüchel, Alexander. Binding of the Major Phasin, PhaP1, from Ralstonia eutropha H16 to Poly(3-Hydroxybutyrate) Granules. Journal of bacteriology, vol.190, no.8, 2911-2919.

상세보기
Coquel, Anne-Sophie, Jacob, Jean-Pascal, Primet, Mael, Demarez, Alice, Dimiccoli, Mariella, Julou, Thomas, Moisan, Lionel, Lindner, Ariel B., Berry, Hugues. Localization of Protein Aggregation in Escherichia coli Is Governed by Diffusion and Nucleoid Macromolecular Crowding Effect. PLoS computational biology, vol.9, no.4, e1003038-.

상세보기
Neeli-Venkata, Ramakanth, Martikainen, Antti, Gupta, Abhishekh, Goncalves, Nadia, Fonseca, Jose, Ribeiro, Andre S.. Robustness of the Process of Nucleoid Exclusion of Protein Aggregates in Escherichia coli. Journal of bacteriology, vol.198, no.6, 898-906.

상세보기
Lindner, Ariel B., Madden, Richard, Demarez, Alice, Stewart, Eric J., Taddei, François. Asymmetric segregation of protein aggregates is associated with cellular aging and rejuvenation. Proceedings of the National Academy of Sciences of the United States of America, vol.105, no.8, 3076-3081.

상세보기
Proenca, Audrey M., Rang, Camilla Ulla, Buetz, Christen, Shi, Chao, Chao, Lin. Age structure landscapes emerge from the equilibrium between aging and rejuvenation in bacterial populations. Nature communications, vol.9, no.1, 3722-.

상세보기
Biotechnol. Lett. Lee S. Y. 1247 16 1994 10.1007/BF00149626 Suppression of filamentation in recombinant Escherichia coli by amplified FtsZ activity S. Y. Lee, Suppression of filamentation in recombinant Escherichia coli by amplified FtsZ activity. Biotechnol. Lett. 16, 1247-1252 (1994).

상세보기
Jiang, X.R., Chen, G.Q.. Morphology engineering of bacteria for bio-production. Biotechnology advances, vol.34, no.4, 435-440.

상세보기

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