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Contenuto archiviato il 2024-04-15

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

Obiettivo

POTENTIALLY, SACCHAROLYTIC CLOSTRIDIA MAY BE EMPLOYED BOTH IN THE LARGE SCALE CONVERSION OF AGRICULTURAL WASTES AND SURPLUSES INTO FUELS AND SOLVENTS, AND IN THE PRODUCTION OF CHIRAL COMPOUNDS BY STEREOSPECIFIC REDUCTIONS WHICH ARE DIFFICULT TO ACCOMPLISH CHEMICALLY. BESIDES FINDING MAJOR NEW USES FOR THE CONSIDERABLE AGRICULTURAL WASTES AND SURPLUSES OF THE EUROPEAN COMMUNITY, PRODUCTS FORMED RELY ON BIOMASS WHICH, IN CONTRAST TO PRESENTLY USED FOSSIL FUELS, IS BOTH RENEWABLE AND ENVIRONMENTALLY ACCEPTABLE.
THE BIOTECHNOLOGICAL POTENTIAL OF THE GENUS CANNOT BE FULLY EXPLOITED UNTIL RELIABLE METHODS HAVE BEEN DEVELOPED FOR THE TRANSFER OF GENETIC INFORMATION INTO AND BETWEEN THEM. THE RESEARCH OF THIS CONTRACT IS AIMED AT DEVELOPING SUCH ENABLING TECHNOLOGY FOR CLOSTRIDIUM ACETOBUTYLICUM, THE ORGANISM OF CHOICE IN THE PRODUCTION OF ACETONE/BUTANOL. THE TECHNOLOGY DEVELOPED SHOULD FACILITATE A RATIONAL APPROACH TO THE IMPROVEMENT OF INDUSTRIAL FERMENTATIONS BY THE GENETIC MANIPULATION OF GENES INVOLVED IN SUBSTRATE UTILISATION AND PRODUCT FORMATION.
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.
THE CONSTRUCTION OF PLASMID VECTORS TO FACILITATE THE TRANSFER AND SUBSEQUENT EXPRESSION OF HETEROLOGOUS GENES IN CLOSTRIDIUM ACETOBUTYLICUM, WITH A VIEW TO IMPROVING SOLVENTOGENESIS.

IN PARTICULAR:
1. A STUDY OF THE REGIONS NECESSARY FOR THE REPLICATION OF CLOSTRIDIAL PLASMIDS IN CLOSTRIDIAL HOSTS AND BACILLUS SUBTILIS.
2. THE CONSTRUCTION OF SHUTTLE VECTORS FOR USE IN E.COLI B.SUBTILIS CLOSTRIDIA AND S.LACTIS.
3. IMPROVEMENT IN THE EFFICIENCY OF C.ACETOBUTYLICUM TRANSFORMATION.
4. CONSTRUCTION OF AN EXPRESSION CARTRIDGE BASED ON THE CLOSTRIDIUM PASTEURIANUM FERREDOXIN GENE PROMOTER.
5. TRANSFER OF HETEROLOGOUS GENES TO C.ACETOBUTYLICUM.

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Coordinatore

Centre for Applied Microbiology and Research (CAMR)
Contributo UE
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Indirizzo
Porton Down
SP4 0JG Salisbury
Regno Unito

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