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Mechanism of lipid-body formation in prokaryotes: how bacteria fatten up (2005)

Wältermann, Marc ; Hinz, Andreas ; Robenek, Horst ; [et al.]

Oxford, UK : Blackwell Science Ltd

Molecular microbiology 55 (2005), S. 0

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ISSN: 1365-2958

Source: Blackwell Publishing Journal Backfiles 1879-2005

Topics: Biology , Medicine

Notes: Neutral lipid accumulation is frequently observed in some Gram-negative prokaryotes like Acinetobacter sp. and most actinomycetes, including the pathogenic Mycobacterium tuberculosis and antibiotic producing streptomycetes. We examined the formation of wax ester- and triacylglycerol (TAG)-bodies in Acinetobacter calcoaceticus and Rhodococcus opacus using microscopic, immunological and biophysical methods. A general model for prokaryotic lipid-body formation is proposed, clearly differing from the current models for the formation of lipid inclusions in eukaryotes and of poly(hydroxyalkanoic acid) (PHA) inclusions in prokaryotes. Formation of lipid-bodies starts with the docking of wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) to the cytoplasm membrane. Both, analyses of in vivo and in vitro lipid-body synthesis, demonstrated the formation of small lipid droplets (SLDs), which remain bound to the membrane-associated enzyme. SLDs conglomerated subsequently to membrane-bound lipid-prebodies which are then released into the cytoplasm. The formation of matured lipid-bodies in the cytoplasm occurred by means of coalescence of SLDs inside the lipid prebodies, which are surrounded by a half-unit membrane of phospholipids.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1111/j.1365-2958.2004.04441.x

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A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds (1999)

Spiekermann, Patricia ; Rehm, Bernd H. A. ; Kalscheuer, Rainer ; [et al.]

Springer

Archives of microbiology 171 (1999), S. 73-80

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ISSN: 1432-072X

Keywords: Key wordsAcinetobacter calcoaceticus ; Azotobacter vinelandii ; Pseudomonas putida ; Ralstonia eutropha ; Rhodococcus opacus ; Nile red ; Nile blue A ; Viable colony staining ; Polyhydroxyalkanoic acid ; Triacylglycerols ; Lipid inclusions ; Poly(3-hydroxybutyric acid) ; Poly(4-hydroxybutyric acid) ; Poly(3-hydroxyoctanoic acid)

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract The oxazine dye Nile blue A and its fluorescent oxazone form, Nile red, were used to develop a simple and highly sensitive staining method to detect poly(3-hydroxybutyric acid) and other polyhydroxyalkanoic acids (PHAs) directly in growing bacterial colonies. In contrast to previously described methods, these dyes were directly included in the medium at concentrations of only 0.5 μg/ml, and growth of the cells occurred in the presence of the dyes. This allowed an estimation of the presence of PHAs in viable colonies at any time during the growth experiment and a powerful discrimination between PHA-negative and PHA-positive strains. The presence of Nile red or Nile blue A did not affect growth of the bacteria. This viable-colony staining method was in particular applicable to gram-negative bacteria such as Azotobacter vinelandii, Escherichia coli, Pseudomonas putida, and Ralstonia eutropha. It was less suitable for discriminating between PHA-negative and PHA-positive strains of gram-positive bacteria such as Bacillus megaterium or Rhodococcus ruber, but it could also be used to discriminate between wax-ester- and triacylglycerol-negative and -positive strains of Acinetobacter calcoaceticus or Rhodococcus opacus. The potential of this new method and its application to further investigations of PHA synthases and PHA biosynthesis pathways are discussed.