Engineering of the bacillus protein secretion apparatus for the effficient production of proteins

Objective

Bacillus species have several advantages as a host for the industrial production of proteins, including high secretion capacity, excellent fermentation characteristics and lack of toxicity. The aims of this project are to construct engineered strains and to design production strategies for the efficient secretion of native enzymes and heterologous proteins from the Bacillus subtilis microbial cell factory. These pre-competitive objectives will be achieved using an inclusive approach in which the efficiency of all stages of the secretion process (early, middle and late) will be evaluated. Bacillus species have a proven track record for the production of native industrial proteins, and extending their product range to heterologous proteins will have highly significant commercial benefits.
Having successfully identified and characterised many of the key components of the secretory pathway, the European Bacillus Secretion Group now proposes to engineer these components for enhanced secretion activity. In some cases this will involve the over-expression of components that form bottlenecks to highlevel secretion. In other cases, individual components will need to be modified so that they can form more productive interactions (or alternatively so that they do not form unproductive interactions) with heterologous proteins. A detailed biochemical study of the interactions between components of the translocase complex, and between components of this complex and secretory proteins, will provide much of the basis for the rational design of engineering strategies.
Another important component of the proposal is a systematic investigation of the expression of the genes for secretory components during exponential and post-exponential growth phases. The identification of links between gene expression, growth, cell physiology and secretion will form the basis for the most important deliverable to be obtained from the proposed research, namely the development of commercially-relevant production strategies for high-level secretion of a wide-range of proteins from engineered Bacillus species.

Programme(s)

• FP4-BIOTECH 2 - Specific research, technological development and demonstration programme in the field of biotechnology, 1994-1998

Topic(s)

• 010101 - Microbial factories

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

UNIVERSITY OF GRONINGEN

Address

30,Kerklaan 30
9750 AA Haren Gn
Netherlands

Participants (7)

Centre National de la Recherche Scientifique

France

Address

2,Place Jussieu
75251 Paris

Forschungszentrum Jülich GmbH

Germany

Address

Wihelm-johnen-straße
52405 Jülich

GENENCOR INTERNATIONAL BV

Netherlands

Address

139,Lange Kleiweg 139
2288 GJ Rijswijk Zh

National Public Health Institute

Finland

Address

166,Mannerheimintie
00300 Helsinki

Novo-Nordisk A/S

Denmark

Address

Novo Alle
2880 Bagsværd

UNIVERSITY OF NEWCASTLE UPON TYNE

United Kingdom

Address

Framlington Place
NE2 4HH Newcastle - Upon Tyne

Universität Bayreuth

Germany

Address

30,Universitätsstrasse
95440 Bayreuth