BioMatNet Item: QLK3-2002-01940 - Eukaryotic polyketides in surrogate hosts (EUKETIDES)

[BioMatNet Database - FP5 Quality of Life Programme]

QLK3-2002-01940
Eukaryotic polyketides in surrogate hosts (EUKETIDES)

Contacts

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Quality of Life - 3. The Cell Factory

Type of Project	Shared Cost Research
Contract No	QLK3-2002-01940
Total Cost
EC Contribution	2,000,000 EUR
Start Date	01-11-2002
Duration	48 Months

Abstract

Eukaryotic organisms contain genes encoding the biosynthesis of complex compounds often with useful biological broperties. EUKETIDES will innovate by obtaining genes directly from a variety of unculturable eukaryotes using new genetic tools. A range of polyketide synthase genes will be isolated and used for production of biologically active molecules by expression in well characterised bacteria and yeast. These new biosynthetic genes will be combined with known modifying genes for the directed biosynthesis of small libraries of compounds, resulting in new technology for producing drugs. Overall we will exploit advances in genetics, chemistry and fermentation technology to capitalise on leading EU science for the production of high-value new compounds.

Objectives

The objectives are:

to harness the biodiversity of biosynthetic activities in hitherto unexploited resources of uncultured and microbial eukaryotes.
to develop systems with combinatorial capacity for biosynthesis of natural small molecules, polyketides in particular.
a significant contribution to developing environmentally friendly production of compounds of high value to the chemical and pharmaceutical industries.

EUKETIDES will yield new polyketides synthesised by diverse and novel fungal type I polyketide synthases widely present in fungi, lichens and other microbial and uncultured eukaryotes. The general approach and the technology developed will allow synthesis of libraries of new compounds, and it will establish combinatorial methodology to increase product diversity. The products and technology platform will be offered to the European pharmaceutical and chemical industry and it will find immediate application in the participating SME.

Activities

Scientific and industrial skills in seven separate European locations with state-of-the art know-how and facilities will join forces in EUKETIDES. Twenty-four or more selected species of uncultured and microbial eukaryotes will be screened for type and diversity of polyketide synthase (PKS) genes and polyketides produced. Each species may contain six or more different PKS genes of approximately 8 kb each and from these a total of six representative types will be selected for expression and production of polyketide backbones in bacteria and yeast. Due to difficulties in recovering large-size nucleic acids from the sources to be exploited the genes will in most cases be recovered in parts bounded by conserved sequences and assembled. Use of trimmed genes in suitable expression vectors circumvents the often stringent and complex activation circuits needed for native gene expression.

Enzymes modifying polyketides will also be exploited for their ability to create structural diversity. Strains of the yeast Saccharomyces cerevisae will be engineered and optimised as hosts expressing heterologous eukaryotic PKS activities. Combinations of PKS genes and modifying activities will be expressed both in strains of Streptomyces and in yeast resulting in hundreds of clones which will be screened for production of new polyketides and biological activities. The technology developed will be used by the participating SME and a specific effort will be made for technology transfer to European industries.

Expected Results

EUKETIDES material deliveries consist of a series of at least twenty-four source species nucleic acids, twenty cDNA libraries, twenty partial PKS sequences, six complete eukaryotic PKS genes and three modifying activities functionally cloned in Streptomyces and in yeast and the pylogeny of all genes and partial sequences concerned. Also to be delivered are yeast strains and vectors for expressing eukaryotic PKS genes, optimisation of media and kinetics for polyketide production in yeast, 12 new polyketide producers, structures and biological activities of eight new compounds. Technological platform and intellectual property deliverables will be provided in a series of six reports, a consortium agreement, a project website, a technology implementation plan, patent applications, presentations at public, scientific and business venues and publications in scientific journals. Student and post-doc training is also a large component of EUKETIDES.

Contacts

Coordinator

Olafur ANDRÉSSON / University of Iceland Institute for Experimental Pathology / ICELAND

Participant