Engineering amide bond ligases (ABLs) for industrial pharmaceutical production'

用于工业药品生产的酰胺键连接酶 (ABL) 工程”

• 批准号: 1792702
• 金额: --
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1792702
• 关键词: Engineering; amide; bond; ligases; ABLs

The amide bond is one of the most significant in pharmaceutical chemistry, featuring in a host of important everyday pharmaceuticals for the treatment of ulcers, high cholesterol and pathogenic infections by bacteria and viruses. It is vital therefore that there exist atom efficient and sustainable green chemical methods for the synthesis of pharmaceutical amides. However, industrial synthetic methods for the preparation of amides suffer from the use of complex or hazardous reagents to accomplish their chemistry and generate a large amount of waste. Because of this lack of efficiency, industrial synthetic chemists are increasingly turning towards 'biocatalysis' or 'Industrial Biotechnology' as the preferred method of synthesising molecules for pharmaceutical production. Biocatalysts, such as enzyme or microbes, typically achieve the synthesis of chemical bonds with excellent atom efficiency and selectivity, and Nature is also expert at synthesising amide bonds, which are the major bonds that hold the structure of proteins together. Until now however, biocatalysts for the formation of amide bonds have received little attention for industrial application, even though such enzyme reactions feature at the top of the list for many chemists looking for biocatalytic solutions to synthetic problems. This is because biocatalytic methods for amide bond formation in Nature, while efficient, are often complex, and difficult to apply out of their natural context. A recently discovered group of enzymes, which we have called amide bond ligases (ABLs), offers new and unexplored promise for biocatalytic amide bond formation, as their reaction chemistry is comparatively simple, and also because the kind of amide bonds that they form, are much more closely related to molecules of real pharmaceutical interest than has previously been the case. In this project, which is a collaboration between biochemists and synthetic chemists at York, and in association with GSK, we propose to thoroughly investigate the synthetic potential of the new ABL enzymes. First we will define the potential and limitations of the natural enzymes using a mixture of synthetic chemistry and biocatalysis. We will then solve the as-yet-undetermined structures of the ABL enzymes using X-ray crystallography and use rational enzyme redesign to expand the potential of the enzymes to catalyse the synthesis of a much wider range of pharmaceutically relevant molecules. We will also use contemporary protein evolution techniques to adapt the enzymes to act on alternative substrates that are of interest to industrial collaborators. This will involve the design and application of new high-throughput screens for amide bond formation to allow us to perform rapid improvement of the enzymes, and also apply new techniques in enzyme cofactor recycling to all

酰胺键是药物化学中最重要的键之一，存在于许多重要的日常药物中，用于治疗溃疡、高胆固醇以及细菌和病毒引起的致病性感染。因此，存在原子效率高且可持续的绿色化学方法来合成药用酰胺至关重要。然而，用于制备酰胺的工业合成方法存在使用复杂或危险试剂来完成其化学反应并产生大量废物的问题。由于缺乏效率，工业合成化学家越来越多地转向“生物催化”或“工业生物技术”作为合成药物生产分子的首选方法。酶或微生物等生物催化剂通常能够以优异的原子效率和选择性实现化学键的合成，而 Nature 也是合成酰胺键的专家，酰胺键是将蛋白质结构保持在一起的主要键。然而，到目前为止，用于形成酰胺键的生物催化剂在工业应用中很少受到关注，尽管这种酶反应对于许多寻求合成问题的生物催化解决方案的化学家来说是首选。这是因为自然界中酰胺键形成的生物催化方法虽然有效，但通常很复杂，并且难以在自然环境之外应用。最近发现的一组酶，我们称之为酰胺键连接酶（ABL），为生物催化酰胺键形成提供了新的、未经探索的前景，因为它们的反应化学相对简单，而且它们形成的酰胺键类型与真正具有药物意义的分子比以前的情况更加密切相关。在这个由约克大学生物化学家和合成化学家合作并与葛兰素史克合作的项目中，我们建议彻底研究新 ABL 酶的合成潜力。首先，我们将结合合成化学和生物催化来定义天然酶的潜力和局限性。然后，我们将使用 X 射线晶体学解决 ABL 酶尚未确定的结构，并使用合理的酶重新设计来扩展酶的潜力，以催化更广泛的药物相关分子的合成。我们还将使用当代蛋白质进化技术来调整酶以作用于工业合作者感兴趣的替代底物。这将涉及设计和应用新的酰胺键形成高通量筛选，使我们能够快速改进酶，并将酶辅因子回收的新技术应用于所有

项目成果

The Broad Aryl Acid Specificity of the Amide Bond Synthetase McbA Suggests Potential for the Biocatalytic Synthesis of Amides.

• DOI: 10.1002/anie.201804592
• 发表时间: 2018-09-03
• 期刊: Angewandte Chemie (International ed. in English)
• 作者: Petchey M;Cuetos A;Rowlinson B;Dannevald S;Frese A;Sutton PW;Lovelock S;Lloyd RC;Fairlamb IJS;Grogan G
• 通讯作者: Grogan G