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 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.
Vidal-Mas
J
J
Department de Microbiologia i Parasitologia Sanitàries, Facultat de Farmacia, Universitat de Barcelona, Spain.
Resina-Pelfort
Haba
E
E
Comas
J
J
Manresa
A
A
Vives-Rego
J
J
D003296:Cooking; D003470:Culture Media; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism*; D006017:Glycolipids / Q000378:metabolism; D008854:Microscopy, Electron; D010078:Oxazines / Q000378:metabolism*; D010938:Plant Oils; D011091:Polyesters / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000254:growth & development* / Q000378:metabolism; D013997:Time Factors
D003470:Culture Media; D005456:Fluorescent Dyes; D006017:Glycolipids; D010078:Oxazines; D010938:Plant Oils; D011091:Polyesters; C418382:rhamnolipid; C044808:nile red