Rapid flow cytometry : Nile red assessment of PHA cellular content and heterogeneity in cultures of Pseudomonas aeruginosa 47T2 (NCIB 40044) grown in waste frying oil

The accumulation of cytoplasmic polyhydroxyalkanoates (PHAs) and the heterogeneity ofbacterial populations were analysed by flow cytometry and SYTO-13 and Nile red staining in rhamnolipid-producing Pseudomonas aeruginosa cultures grown in waste frying oil as carbon source. A combination of SYTO-13 a...

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Bibliographic Details
Published in:Antonie van Leeuwenhoek 2001-10, Vol.80 (1), p.57-63
Main Authors: VIDAL-MAS, J, RESINA-PELFORT, O, HABA, E, COMAS, J, MANRESA, A, VIVES-REGO, J
Format: Article
Language:English
Subjects:
Bacteria
Biological and medical sciences
Biotechnology
Carbon sources
Cooking
Culture Media
Energy
Flow cytometry
Flow Cytometry - methods
Fluorescent Dyes - metabolism
Fundamental and applied biological sciences. Psychology
Glycolipids - metabolism
Heterogeneity
Industrial applications and implications. Economical aspects
Microbiology
Microscopy, Electron
Nile red
Oxazines - metabolism
Plant Oils
Polyesters - metabolism
Polyhydroxyalkanoates
Pseudomonas aeruginosa
Pseudomonas aeruginosa - growth & development
Pseudomonas aeruginosa - metabolism
Rapid flow
rhamnolipids
Time Factors
Use of microorganisms or biopolymers in fossil fuel recovery
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Summary:The accumulation of cytoplasmic polyhydroxyalkanoates (PHAs) and the heterogeneity ofbacterial populations were analysed by flow cytometry and SYTO-13 and Nile red staining in rhamnolipid-producing Pseudomonas aeruginosa cultures grown in waste frying oil as carbon source. A combination of SYTO-13 and Nile red fluorescence with cytometric forward and side scatter values may allow increases in the final production of polyhydroxyalkanoates (PHA) by two basic mechanisms: (i) rapid assessment of polyhydroxyalkanoate content and (ii) definition of flow cytometric cell sorting protocols to select high polyhydroxyalkanoate (PHA)-producing strains. We report a rapid (less than 30 min) flow cytometric assessment of PHAs in Pseudomonas aeruginosa 47T2 following Nile red staining: (i) to estimate cellular PHAs content; (ii) to study heterogeneity of the batch cultures producing PHAs and (iii) to establish the basis for sorting sub-populations with a high capacity to accumulate PHAs.
ISSN:0003-6072
1572-9699
DOI:10.1023/a:1012208225286