QLK3-2001-00435 Solvent tolerant bacteria allowing a broader performance of biotransformations of organic compounds in two phases (BARTOLO)

Contacts




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Biological Conversion : Fine Chemicals : Quality of Life - 3. The Cell Factory

Type of Project	Research Project
Contract No	QLK3-2001-00435
Total Cost
EC Contribution	536,748 EUR
Start Date	01-01-2002
Duration	36 Months

Abstract

BARTOLO is a European solving initiative directed to the acquaintance of knowledge about the unusual tolerance of a Pseudomonas putida strain to solvents with log POW around 2.5, and the exploitation of this property, together with the easy genetic manipulation of the micro-organism, to synthesise a range of fine chemicals using the two-liquid-phase biotransformation technology.

BARTOLO will also consider the scale-up of the processes together with downstream processing and safety. BARTOLO will cover the complete development of a biotechnological process using two-phase biotransformations from molecular biology to product recovery.

Objectives

BARTOLO will focus on the application of a solvent tolerant strain of Pseudomonas putida for an efficient production of a series of fine chemicals using the two-liquid-phase biotransformation technology. The main targets are:

• to study the molecular biology and physiology of the mechanisms allowing the Pseudomonas putida DOT-T1E strain to adapt to solvents with log POW around 2.5.
• to use this micro-organism to synthesise, in optimal conditions, a series of added-value products, i.e. catechols, nitrobenzaldehydes, nitrobenzoates and derivatives of picolinic acid and pyridines.
• to develop methods for the efficient recovery of products in safe downstream processes.

Activities

The research activities in this project correspond to the three following sets of activities:

• molecular bases of solvent tolerance in Pseudomonas putida DOT-T1E. We will unequivocally establish function/dysfunction links in solvent tolerance through the isolation of solvent-sensitive mutants, cloning of the mutations, rescue of the wild-type genes and complementation of mutants. In addition, we will examine conditions that promote the best efflux of substrates/products to avoid accumulation of substrates and products in cell membranes.
• exploiting the catabolic segments of the TOL plasmid for the biosynthesis of catechols, nitrobenzoates, nitrobenzaldehydes, picolinic acid and pyridine. This will involve the construction of suitable mutants and/or the cloning of the suitable catabolic segments on the host chromosome to improve genetic stability and performance predictability.
• optimisation of the performance of the producing strains in reactors, as well as methods for the efficient and safe recovery of products of interest.

Deliverables

This project will provide useful strains and methods for the production of added-value products in two-liquid-phase reactors. The products to be synthesised (catechols, nitrobenzaldehydes, picolinic acid and pyridine derivatives) are useful for the synthesis of vasodilators, vitamins and fungicides. The project will also involve the development of new emulsion reactors. This project will contribute toward the social acceptance of biotechnology showing how molecular biology tailoring can result in an efficient and safe production of chemicals of interest.

Contacts

Coordinator

• Juan L RAMOS / Estación Experimental del Zaidín Dept. of Biochemistry and Molecular / SPAIN

Participant

• Andreas SCHMID / Institute of Biotechnology SWITZERLAND
• MedPlant Genetics, S.L. Environmental Biotechnology / SPAIN
• Hermann J HEIPIEPER / UFZ-Centre for Environmental Research Dept. of Remediation Research / GERMANY