Article (Scientific journals)
Scale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Brognaux, Alison; Francis, Frédéric; Twizere, Jean-Claude et al.
2014In Bioprocess and Biosystems Engineering
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Keywords :
Escherichia coli; extracellular proteome; membrane permeability; scale-up; substrate heterogeneities; scale-down
Abstract :
[en] 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 2D-DIGE analysis (2-Dimensional Differential in-Gel Electrophoresis) 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.
Disciplines :
Biotechnology
Author, co-author :
Brognaux, Alison ;  Université de Liège - ULiège > Gembloux Agro Bio-Tech / Département de chimie et bio-industries > Unité de bio-industries
Francis, Frédéric  ;  Université de Liège - ULiège > Sciences agronomiques > Entomologie fonctionnelle et évolutive
Twizere, Jean-Claude  ;  Université de Liège - ULiège > Chimie et bio-industries > Biologie cell. et moléc.
Thonart, Philippe ;  Université de Liège - ULiège > Département des sciences de la vie > Biochimie et microbiologie industrielles
Delvigne, Frank  ;  Université de Liège - ULiège > Chimie et bio-industries > Bio-industries
Language :
English
Title :
Scale-down effect on the extracellular proteome of Escherichia coli: correlation with membrane permeability and modulation according substrate heterogeneities
Publication date :
14 February 2014
Journal title :
Bioprocess and Biosystems Engineering
ISSN :
1615-7591
eISSN :
1615-7605
Publisher :
Springer, Germany
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]
Available on ORBi :
since 20 December 2013

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