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.

Fields of science

• engineering and technology › industrial biotechnology › metabolic engineering
• natural sciences › biological sciences › molecular biology › molecular genetics
• medical and health sciences › medical biotechnology › genetic engineering › gene therapy
• medical and health sciences › basic medicine › pharmacology and pharmacy › pharmaceutical drugs › antibiotics
• natural sciences › biological sciences › microbiology

Programme(s)

• FP5-LIFE QUALITY - Specific Programme for research, technological development and demonstration on "Quality of life and management of living resources", 1998-2002

Topic(s)

• 1.1.1.-3. - Key action The "Cell factory"

Call for proposal

Data not available

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