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Exploitation of a novel sec-independent secretion pathway for protein production

Objective

Bacteria export numerous proteins across the plasma membrane and this process has been heavily exploited for protein production on an industrial scale. However, the standard 'general secretory pathway' transports proteins in an unfolded state and is inapplicable for the export of the majority of cytosolic and heterologous proteins because of their tendency to fold prematurely. Recent studies, primarily by this partnership, have revealed the operation of a novel bacterial protein export system that recognises signal pepetides bearing twin-arginine motifs (RR-SIGNAL PEPTIDES) and which has the remarkable ability to export large, fully-folded proteins. Critically, RR-signal pepetides direct the export of several heterologous proteins and this application is aimed at the exploitation of this twin-arginine translocation (Tat) system to export active, high value-added proteins from Escherichia coli and Bacillus subtilis. We propose to engineer RR-signal peptides capable of directing highly specific and efficient export in these organisms, and to produce super-exporting stains capable of exporting heterolohous proteins on a commercial scale.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

NETHERLANDS RESEARCH SCHOOL FOR ASTRONOMY - LEGALLY REPRESENTED BY UNIVERSITY OF GRONINGEN
Address
5,Broerstraat 5
9712 CP Groningen
Netherlands

Participants (7)

ALBERT-LUDWIGS-UNIVERSITAET FREIBURG
Germany
Address
Fahnenbergplatz
79085 Freiburg (In Breisgau)
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
France
Address
Cen-saclay
91191 Gif Sur Yvette
GENENCOR INTERNATIONAL BV
Netherlands
Address
139,Lange Kleiweg 139
2288 GJ Rijswijk Zh
JOHN INNES CENTRE
United Kingdom
Address
Norwich Research Park, Colney
NR4 7UH Norwich
THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
United Kingdom
Address
University Offices, Wellington Square
Oxford
UNIVERSITY OF WARWICK
United Kingdom
Address
Gibbet Hill Road
Coventry
University of East Anglia
United Kingdom
Address
University Plain
NR4 7TJ Norwich