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.
TRANSFORMATION OF THE THERMOPHILIC BACTERIUM CLOSTRIDIUM THERMOHYDROSULFURICUM FOR ALLOWING GROWTH ON CELLULOSE AND FOR INCREASING THE TOLERANCE TO ETHANOL.

IN PARTICULAR :

1. EFFICIENT PROCEDURES FOR TRANSFORMATION OF CLOSTRIDIUM THERMOHYDROSULFURICUM WILL BE DEVELOPED.
2. THERMOSTABLE CLONING VECTORS DERIVED FROM CLOSTRIDIAL PLASMIDS WILL BE CONSTRUCTED.
3. GENES FOR POLYSACCHARIDE DEGRADATION FROM THERMOPHILIC CLOSTRIDIA WILL BE TRANSFERED TO CLOSTRIDIUM THERMOHYDROSULFURICUM.
4. THE ETHANOL TOLERANCE OF CLOSTRIDIUM THERMOHYDROSULFURICUM WILL BE INCREASED AND COMPETING PATHWAYS WILL BE ELIMINATED.