Knowledge Based Bio-Economy

LAPTOP

Production optimisation of a new antibiotic

Project Acronym: LAPTOP

Title of project: Lantibiotic production: technology, optimization and improved process

Research area: Biotechnologies (Novel industrial microorganisms with optimised metabolic pathways)

Contract No: 245066

EU Contribution: 2775 000 EURO

Start date: July 2010

Duration: 36 Months

Objectives

The focus of this proposal is to develop an economically viable production process for the lantibiotic NAI-107, a new antibiotic with the potential to treat life-threatening infections caused by multidrug-resistant Gram-positive pathogens. NAI-107 is produced by fermentation using the actinomycete Microbispora sp. It is undergoing formal toxicology studies through NAICONS, an SME coordinating the project that is developing NAI-107 in collaboration with a pharmaceutical company. A challenge in advancing a new antibiotic into clinical development is to devise a production process that will deliver a high quality compound at reasonable yields. This is particularly relevant for NAI-107 since no lantibiotics are industrially produced as drugs for human use and there are no examples of industrial use of Microbispora. The development of a robust and economically feasible production process for NAI-107 requires the integration of basic knowledge of the physiology of the strain which can be best obtained by a combination of classical and post-genomic approaches (proteome/transcriptome), with a detailed knowledge of the production process and its scalability to industrial level.

Expected Impact

This work could lead to a significant advance in the commercial launch of a new type of clinical antibiotics that are effective against multi-drug resistant Gram-positive bacteria that cause particular problems in hospitals resulting in the need to apply extensive hygiene related procedures. Their effective use may be anticipated since one such biologically active peptide, nisin, is already used in food products. This is obtained from cultures of Lactoccus lactis grown on natural substrates, such as milk or glucose and may be used in foods such as cheese, processed meats and beverages during production to extend shelf life by suppressing Gram-positive spoilage and pathogenic bacteria. In the EU where nisin is recognised as E234, it has yet to encounter significant bacterial resistance. However, in order for NAI-107 to reach the market place and have this impact will require considerable research as described here, in addition to further product development. This includes evaluation of it potency and safety through clinical trials leading to selection by a pharmaceutical company as an economically viable product to invest in. At present a wide range of lantibiotics are under investigation with new members of the class being discovered at an increasing rate while some resistance mechanisms, such as immune mimicry, have been identified. There are also questions relating to the use (bacterial targets) and mechanisms of application in therapeutic use. Studies to date show some are poorly absorbed via oral, transdermal or pulmonary routes suggesting they may be preferable for topical use. These factors may be taken into account by pharmaceutical companies when considering the compound of choice for such investment.

Expected Results

Lantibiotics, an abbreviation for lanthionine-containing peptide antibiotics, are a class of peptide antibiotics one of which (designated NAI-107) has been selected for further development by this project. The partners will produce information leading to elucidation of an optimized metabolic pathway for its production. They will also isolate or engineer an enhanced strain of the bacteria producing high levels of the required product as well as designing an improved production medium, fermentation system and an efficient process to recover the protein. Taking these results together the project should produce an efficient production process. The bacteria will be improved through flux analyses and 2D-maps in order to discover proteins involved in primary metabolism up-regulated during antibiotic production. This will be combined with a study of other limiting steps, such as precursor uptake, product excretion and the intrinsic resistance of the producing strain. It will also utilise the results of analysis of the transcriptional regulation of the NAI-107 biosynthetic genes enabling bottlenecks in production to be identified and bypassed by metabolic engineering.

Website of project: www.jic.ac.uk/laptop/

Contacts:

Coordinator: Margherita Sosio, msosio@naicons.com

Organisation: New Anti-Infective Consortium, www.naicons.com

Partners:

Technical University of Denmark, Denmark, www.dtu.dk

University of Palermo, Italy, www.unipa.it

Gnosis SPA, Italy, www.gnosis-bio.com

John Innes Centre, www.jic.bbsrc.ac.uk

University Bonn, Germany, www3.uni-bonn.de

Eberhard Karls University Tuebingen, Germany, www.uni-tuebingen.de