DEVELOPMENT OF GENETIC METHODS FOR TRANSFERING NEW GENETIC INFORMATION INTO THE ANAEROBIC MESOPHILE AND THERMOPHILIC CLOSTRIDIUM SPECIES OF INDUSTRIAL INTEREST. IT IS AIMED AT EXTENDING THE RANGE OF SUBSTRATE UTILIZATION AND AT INCREASING FERMENTATIVE ETHANOL PRODUCTION.
Genetic technologies have been developed for use in lactic acid bacteria and genes have been analysed for key properties that have potential for application in industry. Perfection of genetic engineering technology focussed on vectors, food compatible selection systems, chromosomal integration, gene expression and product secretion. Natural gene transfer processes including conjugation and transposition have also been studied; lactic acid bacteria proteinases are central to flavour generation in food fermentation and, accordingly, were characterized in detail.
Bacteriophage attack is a major industrial problem and the research included extensive in depth analysis of bacteriophages and their mechanisms of resistance.
Heterologous genes were expressed and secreted in Lactococcus. Integration and amplification of genes in the bacterial chromosome were achieved. Lactose catabolism, proteolysis and naturally occurring conjugation were analysed in detail. Hybrid proteinases and protein engineered proteinases were constructed and genes for a lactococcal peptidase and bacteriocin production were characterized. A molecular taxonomy for bacteriophages of lactic acid bacteria was developed and the gene for a bacteriophage lysin was characterized and exploited for a novel flavour generation technology. Bacteriophage receptors on lactococci were characterized and genes for resistance to bacteriophage attack were analysed and are being exploited in improved starter strains.
THE RESEARCH WILL BE CARRIED OUT AS PART OF AN INTEGRATED EFFORT BY A GROUP OF FIVE LABORATORIES (DEPT. DAIRY AND FOOD MICROBIOLOGY, CORK, IRL; NAT. INST. RES. DAIRYING, UK; BUNDESANST. F. MILCHFORSCH., KIEL, FRS; NED. INST. RES. DAIRYING, EDE, NL, GENET. INST. UNIV. GRONIGEN, HAREN, NL).
THE MAIN FEATURES ARE THE FOLLOWING :
1. DEVELOPMENT OF CLONING VECTORS ON THE BASIS OF STREPTOCOCCAL PLASMIDS.
2. EXPRESSION OF HOMOLOGOUS AND HETEROLOGOUS GENES IN STREPTOCOCCI.
3. IMPROVEMENT OF GENE TRANSFER METHODS (E.G. TRANSFORMATION).
4. STABILIZATION OF FERMENTATION PROPERTIES BY TRANSFER OF GENES FROM PLASMIDS INTO THE CHROMOSOME.
5. CHARACTERIZATION AND CONSTRUCTION OF BACTERIOPHAGE RESISTANT BACTERIA.
Funding SchemeCSC - Cost-sharing contracts