Hydroxylations with Novel P450 Enzymes
Functionalization of non-activated C-H bonds is one of the major challenges in chemistry yet this reaction-type is crucially important for the initial activation of simple starting molecules. Hydroxylation of C-H bonds can lead directly to the formation of high value chiral compounds in demand as specialty chemicals and pharmaceutical synthons. Cytochrome P450 enzymes remain unsurpassed in their targeted specificity and scope. Consequently, the application of P450 enzymes in synthetic organic chemistry is considered as “potentially the most useful of all biotransformations”. Despite this potential, the application of P450 reactions in industry has been hampered by several technical bottlenecks widely recognized as:
- Access to P450 enzymes with regio- and stereoselective activity on diverse target compounds
- Biocatalysts with high activity and sufficient operational stability
- Efficient redox partner interaction
- Biotransformation processes operating at high substrate concentrations
- Biotransformation processes allowing sufficient oxygen supply.
In order to overcome these limitations a high level of multidisciplinary collaboration is required. This project addresses these bottlenecks by integrating expertise in novel enzyme recruitment, enzyme engineering, whole cell biocatalyst optimization, process development and scale-up. Each partner brings in leading techniques and develops them further to specifically address P450 processes. Examples include transcriptome analysis to identify active P450 genes, bioinformatics guided enzyme engineering to boost enzyme optimization, and high-throughput process development to optimize reactions and facilitate scale-up. The project will initially characterize a wild-type strain collection and then establish a platform for identification and optimization of new recombinant P450 expression systems. A particular focus will be set on the integration of high throughput microscale process development methods with biocatalyst development. This particular strength of the consortium is considered to be a necessary requirement to bring P450 systems into preparative scale. The project will illustrate the applicability of the developed systems at lab-scale for several pharmaceutical targets and at industrial scale with targets from fine chemistry and pharma. The suite of novel enzymes, host and process development platforms created will have broad impact across the fine chemistry, flavors, fragrances and pharma industry sectors. The overall aim of the project is thus to provide platforms of P450 biocatalysts for a range of target substrates with proven applications at industrial scale.
Bundesministerium für Bildung und Forschung (BMBF)
- Prof. Vlada B. Urlacher (coordinator) - Heinrich-Heine University Düsseldorf – Germany
- PD Dr. Stephan Lütz, Novartis Pharma AG, Basel – Switzerland
- Reidun Aukrust, Managing Director - Synthetica AS, Oslo – Norway
- Dr. Martina Micheletti - University College London – UK
- Dr. Dominik Gront - University of Warsaw – Poland
- Dr. Steffen Rupp - Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V., München – Germany
- Dr. Andreas Vogel - c-LEcta GmbH, Leipzig – Germany