title: Sorting of Streptomyces cell pellets using a complex object parametric analyzer and sorter.
abstract: Streptomycetes are filamentous soil bacteria that are used in industry for the production of enzymes and antibiotics. When grown in bioreactors, these organisms form networks of interconnected hyphae, known as pellets, which are heterogeneous in size. Here we describe a method to analyze and sort mycelial pellets using a Complex Object Parametric Analyzer and Sorter (COPAS). Detailed instructions are given for the use of the instrument and the basic statistical analysis of the data. We furthermore describe how pellets can be sorted according to user-defined settings, which enables downstream processing such as the analysis of the RNA or protein content. Using this methodology the mechanism underlying heterogeneous growth can be tackled. This will be instrumental for improving streptomycetes as a cell factory, considering the fact that productivity correlates with pellet size.
authors.affiliation: Microbial Biotechnology, Institute Biology Leiden, Leiden University.
authors.forename: Marloes L C
authors.initials: ML
authors.lastname: Petrus
authors.affiliation: Microbiology, Kluyver Centre for Genomics of Industrial Fermentation, Utrecht University.
authors.forename: G Jerre
authors.initials: GJ
authors.lastname: van Veluw
authors.affiliation: Microbiology, Kluyver Centre for Genomics of Industrial Fermentation, Utrecht University.
authors.forename: Han A B
authors.initials: HA
authors.lastname: Wösten
authors.affiliation: Microbial Biotechnology, Institute Biology Leiden, Leiden University; d.claessen@biology.leidenuniv.nl.
authors.forename: Dennis
authors.initials: D
authors.lastname: Claessen
mesh_terms: D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods; D048372:Streptomyces coelicolor / Q000166:cytology* / Q000254:growth & development