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），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药物和工业相关化学品的重要组成部分。该项目符合BBSRC的战略优先事项，AAc基序在生物化学中至关重要，是肽和蛋白质的关键构建块，并且参与几乎每一个生物过程。AAc的非天然类似物在工业中具有广泛的应用，作为基于肽的药物、抗体药物缀合物的基本组分，以及在小分子的合成中，这是由于它们赋予这些药物增加的效力、非典型构象和代谢抗性的能力。同样，手性AAl是许多药物成分中的关键基序，也可用作生产非天然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，并且仍然是学术界和工业界的主要挑战。鉴于这些分子在生物学和制药学上的重要性，英国工业界对确定新的战略研究有很高的兴趣，以支持开发创新的、绿色的和更可持续的生产路线，以制备高价值的AAc/AAl。与Almac合作，Almac在通过生物催化解决方案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工业生产非天然AAc/AAl的新酶。该项目的目标是：1）开发用于合成非天然AAc和AAl的新生物催化剂（胺解酶和氨/醛裂解酶）。2）开发新的酶用于开发新的生产方法，以可持续和绿色的方式生产非天然AAc和AAl。3)将学术界开发的这些方法转化为工业制造过程4）为该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相结合，自动筛选和工业生产过程。新的胺脱氢酶和裂解酶将利用这些设施进行开发和工程设计和工业平台。AD和氨LY酶将用于将从腈前体获得的酮和不饱和羧酸转化为所需的非天然AAc。腈水解酶还将用于该项目，以原位生产氨作为AD和LY生物催化剂的底物，使该过程更具可持续性。醛LY酶将用于通过形成新的碳-碳键来生产AAl的新酮-醇前体。该项目将允许通过完全生物催化转化从商业上可获得的廉价底物获得非天然的AAc和AAl。生产的AAc/AAl将由Almac商业化，该项目中产生的新生物催化剂将添加到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.