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

Cited 0 time in webofscience Cited 0 time in scopus
  • Hit : 56
  • Download : 0
<jats:p>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 <jats:italic>Cupriavidus necator</jats:italic>, the best-studied native PHA producer, and recombinant <jats:italic>Escherichia coli</jats:italic> harboring <jats:italic>C. necator</jats:italic> 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 <jats:italic>E. coli</jats:italic> show higher density and localization polarity compared with those of <jats:italic>C. necator</jats:italic>, 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 <jats:italic>C. necator</jats:italic> plays a key role in making these differences between <jats:italic>C. necator</jats:italic> and recombinant <jats:italic>E. coli</jats:italic> strains. This study provides spatiotemporal insights into PHA accumulation inside the native and recombinant bacterial cells.</jats:p>
Publisher
NATL ACAD SCIENCES
Issue Date
2021-08
Language
English
Article Type
Article
Citation

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.118, no.31, pp.e2103956118

ISSN
0027-8424
DOI
10.1073/pnas.2103956118
URI
http://hdl.handle.net/10203/287016
Appears in Collection
PH-Journal Papers(저널논문)CBE-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0