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Metabolic engineering of glycopeptide antibiotics: technology, optimization and production

Objective

Biosynthetic pathways for selected glycopeptides will be analysed and optimized to mobilize the full potential of the actinomycete Cell Factory in the development and production of improved antibiotics. Glycopeptides are structurally complex molecules so their accessibility to chemistry is severely limited, but room for improvement has been demonstrated. In order to have an impact on the health of EU citizens, improved antibiotics need to be discovered and effectively developed. The objectives of this proposal are the development of effective tools for the engineering of bacterial producers of glycopeptides, the discovery of improved glycopeptide antibiotics and the rapid development of promising antibiotics through optimization of the production process. To achieve these objectives, this proposal gathers an inter-disciplinary team of academic and industrial partners, recognized leaders in the application of tools such as combinatorial biochemistry, biotransformation, protein engineering, artificial chromosomes, synthetic chemistry, molecular genetics, metabolic flux analysis, high throughput screening and medical microbiology.
Most of the expected objectives and deliverables were achieved. We have
a) completed the DNA sequences of three glycopeptide clusters;
b) produced glycopeptide intermediates by genetic manipulation of producer strains;
c) overexpressed active oxidases and glycosyltransferases;
d) developed a gene transfer system for Nonomuraea and A. mediterranei;
e) established a defined medium for Nonomuraea growth and mapped metabolic fluxes;
f) implemented a HTS program for hybrid glycosyltransferases;
g) generated novel glycopeptides by mutasynthesis;
h) generated detailed information on parameters affecting A40926 production.

Through these achievements, we can now effectively manipulate two glycopeptide producer strains; we have established the feasibility and limitations to produce novel glycopeptides through chemical, enzymatic, biotransformation and genetic approaches; and we have used the information achieved through the project to develop an efficient process for A40926 production.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

BIOSEARCH ITALIA SPA.
Address
Via Regina Giovanna 17
20129 Milano
Italy

Participants (5)

TECHNICAL UNIVERSITY OF DENMARK
Denmark
Address
Dtu, Blgd. 221
2800 Lyngby
THE UNIVERSITY OF TUEBINGEN
Germany
Address
Wilhelmstrasse 5
72074 Tuebingen
UNIVERSITA DEGLI STUDI DI PALERMO
Italy
Address
Piazza Marina, 61
90133 Palermo
UNIVERSITY OF GRONINGEN
Netherlands
Address
4,Nijenborgh 4
9747 AG Groningen
UNIVERSITY OF ZURICH
Switzerland
Address
71,Raemistrasse 71
CH-8091 Zurich