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

该项目是Castagnolo博士，UCL和Almac之间建立合作的结果，旨在开发新的生物催化剂，胺脱氢酶（AD）和裂解酶（LY）酶，用于手性非天然氨基酸（AAc）和氨基醇（AAl）的可持续工业生产，作为制造与工业相关的药物和化学品的基本组成部分。该项目符合BBSRC“工业生物技术的新战略方法”的战略重点。AAc基序在生物化学中具有至关重要的意义，它是多肽和蛋白质的关键组成部分，几乎参与了每一个生物过程。AAc的非天然类似物在工业上有广泛的应用，作为肽类药物、抗体药物偶联物和小分子合成的基本成分，因为它们能够赋予此类药物更高的效力、非典型构象和代谢抗性。同样，手性AAl是许多药物成分中的关键基序，也可以用作生产非天然AAc的前体。手性非天然AAc/AAl的化学合成并不简单，它需要苛刻的反应条件，仍然是学术界和工业界面临的重大挑战。鉴于这些分子在生物和制药方面的重要性，英国工业界对确定新的战略研究有很高的兴趣，以支持创新、绿色和更可持续的制造路线的发展，以制备高价值的AAc/AAl。该项目与Almac公司合作，该公司在通过生物催化溶液生产药物和化学品方面拥有世界领先的经验，该项目旨在开发可用于非天然AAc/AAl工业生产的新酶。该项目的目标是：1)开发新的生物催化剂（胺脱氢酶和氨/醛裂解酶）用于合成非天然AAc和AAl。2)利用新酶开发新的生产方法，以可持续和绿色的方式生产非天然的AAc和AAl。3)将学术界发展的方法应用于工业制造过程；4)为本项目指定的博士生提供可持续化学和工业生物技术的多学科培训，将分子生物学、宏基因组学、计算生物工程和生物化学与化学合成、自动化筛选和工业制造过程相结合。利用Almac现有的设施和工业平台，将开发和设计新的胺脱氢酶和裂解酶。AD和氨LY酶将用于将从腈前体中获得的酮酸和不饱和羧酸转化为所需的非天然AAc。该项目还将使用硝化酶原位生产氨，作为AD和LY生物催化剂的底物，使该过程更具可持续性。醛LY酶将通过形成新的碳-碳键来产生新的AAl酮醇前体。该项目将允许通过完全的生物催化转化，从市售和廉价的底物中获得非天然的AAc和AAl。生产的AAc/AAl将由Almac商业化，项目中产生的新生物催化剂将被添加到Almac生物催化工具箱中并商业化。

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 processes4) 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.