Adaptation of Mycobacterium smegmatis to an industrial scale medium and isolation of the mycobacterial porinMspA

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Show simple item record Wendel, Sebastian O. Perera, Ayomi S. Pfromm, Peter H. Czermak, Peter Bossmann, Stefan H. 2013-07-10T19:59:17Z 2013-07-10T19:59:17Z 2013-07-10
dc.description.abstract The adaptation of the organism to a simple and cost-effective growth medium is mandatory in developing a process for large scale production of the octamericporinMspA, which is isolated from Mycobacterium smegmatis. A fermentation optimization with the minimal nutrients required for growth has been performed. During the fermentation, the iron- and ammonium chloride concentrations in the medium were varied to determine their impact on the observed growth rates and cell mass yields. Common antibiotics to control contamination were eliminated in favor of copper sulfate to reduce costs. MspA has been successfully isolated from the harvested M. smegmatisusing aqueous nOPOE (noctyloligooxyethylene) at 65°C. Because of the extraordinary stability of MspA, it is possible to denature and precipitate virtually all other proteins and contaminants by following this approach. To further purify the product, acetone is used for precipitation. Gel electrophoresis confirmed the presence and purity of MspA. A maximum of 840µg (via Bradford assay) of pure MspA per liter of the optimized simple growth medium has been obtained. This is a 40% increase with respect to the previously reported culture medium for MspA. en_US
dc.language.iso en_US en_US
dc.relation.uri en_US
dc.subject Medium en_US
dc.subject Optimization en_US
dc.subject Mycobacterium en_US
dc.subject Smegmatis en_US
dc.subject MspA en_US
dc.subject Protein extraction en_US
dc.title Adaptation of Mycobacterium smegmatis to an industrial scale medium and isolation of the mycobacterial porinMspA en_US
dc.type Article (publisher version) en_US 2013 en_US
dc.citation.doi doi:10.2174/1874285801307010092 en_US
dc.citation.epage 98 en_US
dc.citation.jtitle The Open Microbiology Journal en_US
dc.citation.spage 92 en_US
dc.citation.volume 7 en_US
dc.contributor.authoreid pfromm en_US
dc.contributor.authoreid pczermak en_US
dc.contributor.authoreid sbossman en_US

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