Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1995-02-24
1997-12-02
Jones, W. Gary
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536 2432, C07H 2104
Patent
active
056937741
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to the area of recombinant DNA technology and in particular to the application thereof to myxobacteria, but preferably to myxobacteria of the Sorangium/Polyangium group, for the localization, identification and cloning of individual genes and even of entire gene clusters within the bacterial genome.
BACKGROUND OF THE INVENTION
The myxobacteria of the Sorangium/Polyangium group are highly specialized organisms which are frequently detectable in soil samples, dead plant material or in animal dung. Characteristic of this group of microorganisms is, inter alia, their ability to utilize cellulose or cellulose-containing degradation products as sole C source. Another characteristic feature of this group is their ability to produce highly active secondary metabolites.
To date, numerous strains of this group of organisms which are able, for example, to synthesize plant-microbicidal compounds have been described. Of particular importance in this connection are the so-called soraphens, whose structural particulars are described in detail, for example, in EP 0 358 606. The soraphens are macrocyclic compounds which have a cytostatic activity and a favorable biocidal spectrum against pathogenic microorganisms, but especially against phytopathogenic fungi. These compounds have very advantageous curative, systemic and, in particular, preventive properties and are therefore outstandingly suitable for use for the therapeutic treatment of mammalians and protecting numerous crop plants.
It is also known of other representatives of the group of myxobacteria that they are able to synthesize highly active compounds with antibiotic and pharmaceutical potential of these compounds and the importance, resulting therefrom, for plant protection, there is great interest in understanding the genetic bases of their synthesis in order thus to provide the possibility of being able to influence them specifically where appropriate.
This is desirable in particular because the natural producer strains, as also in the case of soraphen biosynthesis, very often provide the interesting secondary metabolites only in inadequate concentrations which are far from sufficient to satisfy the demand for substance for wider-ranging activity assays, to say nothing of commercial production.
Furthermore, these strains prove in many cases to be problematic and difficult to handle and therefore unsuitable for use on an industrial scale, for example in a fermentation process.
It would therefore be extremely desirable to decipher the genetic bases of secondary metabolite production in individual interesting representatives of the myxobacteria, because this opens up a large number of possibilities for intervening directly or indirectly in the synthetic process. Thus, for example, it would be conceivable to attempt to increase the productivity of the producer strains by targeted mutations or insertion of further gene copies. Another conceivable alternative relates to a possible transfer of the complete gene cluster which is responsible for secondary metabolite biosynthesis, or else selected parts thereof, into an organism strain which is optimized for production purposes and which can then be employed for metabolite production on the industrial scale.
However, the precondition for this is the provision of a method which makes it possible for these organisms to be subjected to direct and preferably targeted genetic manipulation using recombinant DNA techniques, for example by targeted incorporation of new genes or gene fragments or other DNA sequences, including whole plasmids, into the genome of the myxobacteria.
Individual representatives from the group of myxobacteria have already been the subject of investigations in this direction. Particular interest was directed in this connection primarily at Myxococcus xanthus, a myxobacterium which has to date already been rather well researched and for which various gene transfer methods have also already been described. Thus, for example, the coli phage P1 was ini
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Ligon James M.
Neff Snezana
Schupp Thomas
Atzel Amy
Jones W. Gary
Meigs J. Timothy
Novartis Finance Corporation
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