MEGA-TOP: Metabolic engineering of glycopeptide antibiotics - technology, optimization and production

• Research unit: EU RFP
• Project number: 99.0241

Key words (English)

Vancomycin; biosynthesis; enzyme; cytochrome-P450; teicoplanin

Alternative project number (English)

EU project number: QLK3-1999-00650

Research programs (English)

EU-programme: 5. Frame Research Programme - 1.1.3 The 'cell factory'

Short description (English)

See abstract

Partners and International Organizations (English)

Coordinator: Biosearch Italia SPA, Gerenzano (I)

Abstract (English)

Glycopeptides are an important class of antibiotics, which interfer with cross-linking during the biosynthesis of the bacterial cell wall peptidoglycan. Vancomycin and teicoplanin are currently in clinical use. The aims of this project are to provide new knowledge and technologies to allow the production of improved glycopeptide antibiotics by engineering of naturally occurring biosynthetic pathways to vancomycin and teicoplanin.
The contributions of the Zurich group have been in the following areas:
1) chemical synthesis of putative biosynthetic intermediates, in particular, linear oligopeptides that should be used to study phenol coupling enzymes of the P450 class. A synthetic procedure for preparaing these oligopeptides has now been established using solid-phase methods.
2) overproduction of P450 enzymes that are involved in the coupling of aromatic side-chains during the biosynthesis of the aglycone core structure of the glycopeptide antibiotics. Three P450 enzymes have been identified, that catalyze these three key phenol coupling reactions. These three enzymes have been overproduced in E. coli, and two have been crystallized and their 3D structures have been determined to 1.7 and 1.9Å resolution. Biochemical assays have been peformed to characterize these P450-like proteins. Currently, efforts are underway to determine the correct substrates of these P450-like enzymes, and to study their mechanisms of action. These enzymes may be useful later to transform modified oligopeptides, which in turn may lead to novel glycopeptide antibiotics.

References in databases (English)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 99.0241