Biosynthesis of five-membered heterocyclic rings
Lead Research Organisation:
University of Aberdeen
Department Name: Chemistry
Abstract
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Technical Summary
We propose to investigate the mechanism of the formation of heterocycles by enzyme. These five membered rings are critical components of many important biologically active molecules. We have made significant progress in determining the chemical mechanism, which is the first step to harnessing the enzyme. We intend to pursue the mechanism by novel labeling strategies including stable isotope labeling of substrates. We will also probe the basis of recognition using NMR approaches to identify the crucial residues involved in binding substrate to the enzyme. This is important because in the long term we would wish to replace amino acids by non peptide like groups. We have shown that NMR appears to detect intermediates that form during the reaction. The oxidation state of the rings is important, since even this subtle modification controls activity and stability of the compounds. We have shown that air can be sued to replace in part the enzyme mechanism. We intend to develop further the chemical route but also to study the enzymatic route. There are important and puzzling differences between enzymes that catalyse this reaction. The ability to control the stereocentres of amino acids is also central the activity of these compounds. The mechanism by which residues adjacent to thiazolines are epimerised is unknown. We will determine whether it is spontaneous or enzyme catalysed. We have developed an approach using a combination of peptide synthesis and protein ligation that will allow us incorporate non natural amino acids. This will not only give us exquisite control of the biochemical experiments but lead to more interesting chemical scaffolds in the future.
Planned Impact
Who might benefit from this research? In terms of economic impact the main beneficiaries will be the UK Industrial
Biotechnology sector. We see the technology as enabling new biologically active molecules. The adoption of new 'greener'
biotransformation processes will enable the difficult transformations required for the production of fine chemicals and
pharmaceuticals. The pharmaceutical industry in the UK generates a positive trade balance, and innovation in these
sectors is critical for the success of the UK as a whole. Society will benefit by the production of new pharmaceutical lead
molecules.
How might they benefit from this research? The Industrial Biotechnology sector will benefit from adopting new but de-risked
technology. Understanding this very useful flexible enzyme will give rise to whole new materials we test for bioactivity in a
number of disease targeted screens. These compounds can then be developed for commercial application and can be
licenced or co-developed with industry. There is an urgent need for a diverse arrays of complex molecules to refill the
pharmaceutical drug discovery pipelines. Our approach will produce materials that can be modified easily and thus tuned to
a particular application. New materials with unusual properties will also be produced as part of this work and these may
provide new products or ideas for new products. The production of heterocycles is difficult to achieve synthetically and
often gives low yields despite the use of large quantities of reagents. The use of efficient biotransformation enzymes will
reduce the use of chemical reagents, solvents, energy and waste products.
Biotechnology sector. We see the technology as enabling new biologically active molecules. The adoption of new 'greener'
biotransformation processes will enable the difficult transformations required for the production of fine chemicals and
pharmaceuticals. The pharmaceutical industry in the UK generates a positive trade balance, and innovation in these
sectors is critical for the success of the UK as a whole. Society will benefit by the production of new pharmaceutical lead
molecules.
How might they benefit from this research? The Industrial Biotechnology sector will benefit from adopting new but de-risked
technology. Understanding this very useful flexible enzyme will give rise to whole new materials we test for bioactivity in a
number of disease targeted screens. These compounds can then be developed for commercial application and can be
licenced or co-developed with industry. There is an urgent need for a diverse arrays of complex molecules to refill the
pharmaceutical drug discovery pipelines. Our approach will produce materials that can be modified easily and thus tuned to
a particular application. New materials with unusual properties will also be produced as part of this work and these may
provide new products or ideas for new products. The production of heterocycles is difficult to achieve synthetically and
often gives low yields despite the use of large quantities of reagents. The use of efficient biotransformation enzymes will
reduce the use of chemical reagents, solvents, energy and waste products.
Organisations
Publications
BrĂ¡s NF
(2016)
The Catalytic Mechanism of the Marine-Derived Macrocyclase PatGmac.
in Chemistry (Weinheim an der Bergstrasse, Germany)
Houssen WE
(2014)
An efficient method for the in vitro production of azol(in)e-based cyclic peptides.
in Angewandte Chemie (International ed. in English)
Jaspars M
(2014)
The origins of cyanobactin chemistry and biology.
in Chemical communications (Cambridge, England)
Koehnke J
(2013)
The cyanobactin heterocyclase enzyme: a processive adenylase that operates with a defined order of reaction.
in Angewandte Chemie (International ed. in English)
Koehnke J
(2015)
Structural analysis of leader peptide binding enables leader-free cyanobactin processing.
in Nature chemical biology
Koehnke J
(2014)
The structural biology of patellamide biosynthesis.
in Current opinion in structural biology
Oueis E
(2016)
Enzymatic Macrocyclization of 1,2,3-Triazole Peptide Mimetics.
in Angewandte Chemie (International ed. in English)
Oueis E
(2017)
Bypassing the proline/thiazoline requirement of the macrocyclase PatG.
in Chemical communications (Cambridge, England)
Description | We have engineered enzymes that can make entirely new molecules. The molecules are hydrids of peptides and organic molecules. These molecules have value as drugs and diagnostics. We have formuated an entirely new process for making complex macrocycles. Along the way there are many key enabling findings. These include the characterisation of the kinetics of the enzymes that control modification of peptides including macrocyclisation. We have also presented findings on how the enzymes in the pathway control the timing of the formation of modifications. These findings were crucial in moving to reach the key finding, the making of a new molecule. |
Exploitation Route | We are going to try form a company. The company is called GyreOx. |
Sectors | Healthcare Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
Description | We have been able to solve the structure of a novel protein that may be involved in the epimerisation of amino acids. This is important as controlled epimerisation is chemically very useful in the pharmaceutical industry. Our work has inspired new efforts in peptide drug discovery. We have shown that it is possible to combine organic chemistry to make highly diverse and modifiable macrocycles. This has changed how people think about peptides in medicine. |
First Year Of Impact | 2014 |
Sector | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
Impact Types | Economic |
Title | An engineered heterocyclase |
Description | The engineering of an enzyme that can generate azoline heterocycles from dipeptides containing XC, XS or XT by attaching the precursor peptide leader to the N terminus of the native enzyme thus activating it permanently. |
IP Reference | GB1419650.5 |
Protection | Patent application published |
Year Protection Granted | 2014 |
Licensed | No |
Impact | Start-up company Ripptide formed in Sept 2015, which will formally in-license the IP in July 2016. |
Company Name | GyreOx |
Description | GyreOx develops in vitro technology that enables macrocycle production to be achieved on multiple scales. |
Year Established | 2019 |
Impact | Secured ca £1.1 M investment |
Website | http://www.gyreox.com |
Company Name | Ripptide Pharma |
Description | Ripptide Pharma develops drugs for autoimmune and inflammatory diseases, as well as cancer. |
Year Established | 2015 |
Impact | None yet, we will in-license IP formally in July 2016 |
Website | http://www.ripptide-pharma.com/ |
Description | Build a medicine |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Led to further invitations to have a stand at European Researcher's night and to present to schools liaison officers NA |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.facebook.com/marinebiodiscoverycentre |