NDLI: Scale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according to substrate heterogeneities

Content Provider	Springer Nature Link
Author	Brognaux, Alison Francis, Frédéric Twizere, Jean Claude Thonart, Philippe Delvigne, Frank
Copyright Year	2014
Abstract	Protein leakage is induced in well-mixed fed-batch bioreactor by comparison with cultures carried out in scale-down conditions. This effect is attributed to a progressive increase of cell membrane permeability and the synthesis of several outer-membrane components allowing to cope with substrate limitation commonly found in high cell density culture. A comparative analysis of protein leakage has thus been performed in well-mixed bioreactors and in scale-down devices. The extracellular proteome of E.coli has been investigated by 2D-gel electrophoresis and identified by subsequent MALDI-TOF analysis. On 110 picked spots, 67 proteins have been identified and the sub-localisation and the molecular function of these proteins have been determined. A majority of the extracellular proteome was composed of outer-membrane and periplasmic proteins (64 %) confirming the fact that leakage is involved in high cell density cultures. About 50 % of this extracellular proteome was composed of transport and binding proteins. Furthermore, the more abundant spots on the gel corresponded to porin proteins and periplasmic transporters. In particular, the OmpC porin was found to be very abundant. Moreover, the scale-down effect on this extracellular proteome has been investigated by two-dimensional differential in-gel electrophoresis analysis (2D-DIGE), and significant differences have been observed by comparison with culture carried out in well-mixed systems. Indeed, since substrate limitation signal is alleviated in this kind of apparatus, cell permeability was lowered as shown by flow cytometry. In scale-down conditions, protein leakage was thus less abundant.
Starting Page	1469
Ending Page	1485
Page Count	17
File Format	PDF
ISSN	16157591
Journal	Bioprocess Engineering
Volume Number	37
Issue Number	8
e-ISSN	16157605
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-02-14
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Escherichia coli Extracellular proteome Membrane permeability Scale-up Substrate heterogeneities Scale-down Biotechnology Industrial and Production Engineering Environmental Engineering/Biotechnology Industrial Chemistry/Chemical Engineering Food Science
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Biotechnology

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