Funding tool: Basic research projects
Project-ID: LS11-014
Project start: 01. November 2012
Project end: will follow
Runtime: 36 months / finished
Funding amount: € 299.000,00

Brief summary:

Tapping Bacterial Resources – Accessing Secondary Metabolites of the Uncultivated

A large fraction of microbes cannot be cultivated with the current laboratory methods. These microbes most likely produce an enormous diversity of so far undescribed compounds. Moreover, bacteria often posses biosynthesis gene clusters which are not expressed under standard laboratory conditions. This promises a huge potential for the discovery of novel leads for drug research from these largely untapped resources, which can be made accessible by functional and bioinformatic analysis of genome and metagenome libraries. Antibiotics and antitumor substances are to a large extent either natural products or semisynthetic derivative of natural products; it can therefore be anticipated that the potential for novel lead structure discovery for these classes of pharmaceuticals is high. A major problem in the identification of novel antibiotics is the fact that common active substances have long been described. To discover novel structures and classes of compounds untapped sources as the DNA of uncultivable bacteria are needed. Moreover, the work intensive rediscovery of already well known structures and genes must be prevented.
In the current proposal we plan to establish a platform allowing a high throughput screening (HTS) of (meta)genome libraries, which allows an innovative combination of micropatterning tools, cellular biosensors for detection of active clones and pyrosequencing of (meta)genomic libraries. Capillary force patterning together with antibiotic- and stress- sensitive biosensors will be used to permit a high density screen with high throughput and the detection of active principals in sublethal concentrations. To establish a higher diversity of metabolites different host strains and shuttle vectors will be used. For the verification of first hit results independent assays with microfluid systems will be established. To prevent the re-discovery of already well known metabolites and to predict potential metabolite classes, (meta)genome libraries will be submitted to pyrosequencing. Clones resulting in positive hits in the functional screen will be also sequenced, which will allow a fast classification of the metabolite classes involved. This reduces the further work in the scale-up process and the following identification and isolation of the natural products. The project will allow the establishment of a (meta)genomic compound library (a metachemical library) as a base not only for subsequent antibiotics screening but as a platform for the screening of novel lead structures and genes in general.

Keywords:
metagenomic libraries, high-throughput-screening, capillary force patterning, microfluid systems, antibiotics, secondary metabolites, biosensors, pyrosequencing, biosynthetic cluster in silico prediction