engleski

Generation of novel molecules by targeted and random recombination of biosynthetic pathways

In spite of the introduction of novel technologies in pharmaceutical research (e.g. genomics, high throughput screening, combinatorial and parallel synthesis) the success of these programs is lagging behind the high expectations. This may be partially due to the lack of novel compound classes to be screened for interaction with various targets. Therefore, the interest in natural compound pools has recently increased again. The significant advances in our understanding of the biosynthetic machinery of complex compound classes such as polyketides has led to the introduction of combinatorial genetics, allowing the modification of natural pathways. Targeted manipulation, e.g. of the polyketide synthetases type I (reduced complex polyketides, such as macrolides) and type II (polyaromatic polyketides, such as tetracyclines and anthracyclines), has led to the generation of novel compounds with significant structural diversity as compared to the natural parental compound. In principal the approaches can be divided into targeted and random. The targeted approach allows precise tailoring of known scaffold. However, it requires substantial workload and most often the compounds turn out to be weakly active or inactive for the desired purpose. On the other hand, random approaches such as gene shuffling technology, although aimed for broad derivatisation, are so far limited for the use on a gene level. The present technologies can't be applied on gene clusters or complex pathways. To overcome this bottleneck we are currently evaluating the random approach, which enables the broadest recombination based on the inhibition of mismatch repair system (MRS). MRS consists of various proteins that correct mismatches occurring during transcription and functions as a guardian of genomic integrity. Besides, it prevents recombination of diverged sequences. Inactivation of MRS enables recombination of genes, gene clusters or even the whole genomes that diverge more than 30-50% (isn't it too much?). In this way, it is possible to screen in vivo the generation of novel compounds by combining various pathways. We are now adapting this system for broad modification of secondary metabolite biosynthesis to identify new chemical entities for their application in life sciences rapidly and efficiently.

polyketides; novel compound classes; targeted and random recombination; combinatorial genetics

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