Development of amine-dehydrogenase and lyase biocatalysts for the sustainable manufacturing of unnatural chiral amino acids and amino alcohols

This project comes as the result of an established collaboration between Dr Castagnolo, UCL and Almac and it aims to develop new biocatalysts, amine dehydrogenase (AD) and lyase (LY) enzymes, for the sustainable industrial production of chiral unnatural amino acids (AAc) and amino alcohols (AAl) as essential building blocks in the manufacturing of drugs and chemicals of industrial relevance.
This project is in line with BBSRC's strategic priority in "new strategic approaches to industrial biotechnology".
The AAc motif is of crucial importance in biochemistry, being the key building block of peptides and proteins and being involved in almost every biological process. Unnatural analogues of AAc find broad application in industry, as fundamental components of peptide-based pharmaceuticals, antibody drug conjugates and in the synthesis of small molecules, due to their ability to confer to such drugs increased potency, atypical conformations and metabolic resistance. Similarly, chiral AAl's are key motifs in many pharmaceutical ingredients and can be also used as precursors in the production of unnatural AAc. The chemical synthesis of chiral unnatural AAc/AAl is not trivial, it requires harsh reaction conditions and still represents a major challenge for both academia and industry. Given the biological and pharmaceutical importance of these molecules, the UK industry has high interest to identify new strategic research to underpin the development of innovative, green and mores sustainable manufacturing routes to prepare high-value AAc/AAl.
In collaboration with Almac, that has world leading experience in the manufacturing of drugs and chemicals through biocatalytic solutions, this project aims to develop new enzymes which can be exploited in the industrial production of unnatural AAc/AAl.
The objectives of the project are:
1) To develop new biocatalysts (amine dehydrogenases and ammonia/aldehyde lyases) for the the synthesis of unnatural AAc and AAl.
2) To exploit the new enzymes in the development of new manufacturing methodologies to produce unnatural AAc and AAl in a sustainable and green manner.
3) To translate these methodologies developed in academia into industrial manufacturing processes
4) To offer the PhD student appointed with this project a multidisciplinary training in sustainable chemistry and industrial biotechnology, combining molecular biology, metagenomics, computational bio-engineering and biochemistry with chemical synthesis, automated screening and industrial manufacturing processes.
New amine dehydrogenase and lyase enzymes will be developed and engineered taking advantage of the facilities and industrial platforms available at Almac. The AD and ammonia LY enzymes will be used to convert keto- and unsaturated carboxylic acids, in turn obtained from nitrile precursors, into the desired unnatural AAc. Nitrilase enzymes will also be used in the project to produce in situ ammonia as substrate for AD and LY biocatalysts making the process even more sustainable. The aldehyde LY enzymes will be used to produce, new keto-alcohol precursors of AAl through the formation of new carbon-carbon bonds. This project will allow to obtain unnatural AAc and AAl from commercially available and cheap substrates through entirely biocatalytic transformations. The produced AAc/AAl will be made commercially available by Almac as well as the new biocatalysts generated in the project will be added to the Almac biocatalysis toolbox and commercialised.