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DEVELOPMENT OF HOST/VECTOR SYSTEMS IN CLOSTRIDIA OF INDUSTRIAL AND AGRICULTURAL IMPORTANCE

Objective

DEVELOPMENT OF METHODS FOR TRANSFERRING NEW GENETIC INFORMATION INTO ANAEROBIC MESOPHILIC AND THERMOPHILIC CLOSTRIDIUM SPECIES OF INDUSTRIAL INTEREST. THE INITIAL AIM IS TO EXTEND THE RANGE OF SUBSTRATES UTILIZED AND INCREASE/MANIPULATE THE AMOUNTS OF PRODUCTS (BUTANOL/ACETONE/ETHANOL) FORMED.
One group of microbes, called clostridia, are remarkably adept at producing a wide variety of useful chemicals. Research concentrated on 2 organisms which convert biomass into the chemical biofuels, ethanol and butanol, and acetone and the development of techniques needed to breed improved strains of these microorganisms, which will produce higher yields of biofuels and efficiently use many different sources of biomass.

Selective breeding requires the isolation of favourable genes, procedures for introducing them into the organisms, and vehicles (plasmids) on which they may be carried. 2 systems for introducing genes into Clostridium were developed: an electrical discharge was used to 'punch' holes in the exterior wall of the cell, through which genetic information may enter; use was made of a naturally occurring phenomenon called mating, whereby bacteria exchange genes with each other. Several genes endowing favourable characteristics on microorganisms were also isolated and numerous plasmids constructed for transporting them into clostridia. Using the techniques developed, it was shown that the organism under study can be altered such that it can grow on certain components of plant material (beta-glucan and lichenan) that were hitherto undigestible. The work establishes the feasibility of breeding improved microorganisms for the production of biofuels and industrially important chemicals.
TRANSFER BY CONJUGATION OF GENES CONCERNED WITH SUBSTRATE UTILIZATION AND SOLVENT PRODUCTION IN SACCHAROLYTIC CLOSTRIDIA. TRANSPOSONS WILL BE USED TO IDENTIFY AND ISOLATE GENES OF INTEREST.

IN PARTICULAR :

1- THE EFFICIENT TRANSFER OF TN916 AND TN1545 INTO CLOSTRIDIUM ACETOBUTYLICUM WILL BE ASSURED FOR INSERTIONAL MUTAGENESIS.
2- GENES OF INTEREST, INTO WHICH TN916 OR TN1545 HAS INSERTED, WILL BE ISOLATED BY CLONING IN ESCHERICMIA COLI.
3- INTRODUCTION OF FOREIGN GENES INTO CLOSTRIDIUM ACETOBUTYLICUM BY CONJUGAL TRANSFER OF MOBILIZED PLASMIDS.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

University of Wales, Aberystwyth
Address
Old College King Street
SY23 2AX Aberystwyth
United Kingdom

Participants (2)

Centre for Applied Microbiology and Research (CAMR)
United Kingdom
Address
Porton Down
SP4 0JG Salisbury
Technische Universitaet Muenchen
Germany
Address
Lichtenbergstrasse 4
85748 Garching