Synthetic Biology Pathways to Isoquinoline Alkaloids

异喹啉生物碱的合成生物学途径

• 批准号: BB/G014426/1
• 负责人: John Ward
• 金额: $ 90.88万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG014426%2F1
• 关键词: Synthetic; Biology; Pathways; Isoquinoline; Alkaloids

For thousands of years mankind has been using plants and medicinal compounds from plants for treating pain, diseases, infections and also for recreational purposes. Many of these plant derived compounds have also found their way into modern drug use for example morphine and codeine for pain relief, atropine to increase heart rate and for irritable bowel syndrome, vinblastine to treat cancers and quinine for malaria. There are over 14,000 alkaloid chemical structures known and these represent a wealth of potential biological activity. However many of these alkaloids occur in small amounts or in plants that are rare or only grow in parts of the world that are difficult to access. The pathways that plants use to synthesise alkaloids are often long with many biosynthetic steps. There are some key intermediate compounds that are common to many plant alkaloid pathways and the complexity of alkaloid structures comes from the different later steps in the pathways that different plants have. It would be of great benefit to be able to make some of these key intermediate alkaloid compounds in a more controlled way and to be able to modify these alkaloid structures using enzymes or chemistry. In this way novel biologically active drugs could be found. The pathways for the synthesis of several alkaloids have begun to be determined but many of the steps for some of the more complex alkaloids are still unknown. In the last few years researchers have begun to isolate the enzymes (and their genes) for some of the key steps in the synthesis of some alkaloids. Some of these enzymes carry out very important steps such as the joining together of rather simple compounds derived from simple amino acids. We intend to use the techniques of whole gene synthesis, synthetic biology and pathway design to make some of these key alkaloid synthesising enzymes and put them into a laboratory bacterium. In this way we can combine different plant based and bacterial enzymes and build synthetic pathways for the creation of new chemical compounds that could be screened for useful medicinal activities. By having the pathways in bacteria we would be able to generate them in a reproducible way using a renewable resource and not be dependant on the growth of difficult or rare plants. In addition, extensive chemical transformations using methods with high environmental impact would not be required which are currently used to synthesise such isoquinolines.

几千年来，人类一直在使用植物和植物中的药用化合物来治疗疼痛、疾病、感染，也用于娱乐目的。许多源自植物的化合物也已进入现代药物用途，例如用于缓解疼痛的吗啡和可待因、用于提高心率和治疗肠易激综合症的阿托品、用于治疗癌症的长春花碱以及用于治疗疟疾的奎宁。已知的生物碱化学结构超过 14,000 种，它们代表着丰富的潜在生物活性。然而，许多这些生物碱含量很少，或者存在于稀有植物中，或者仅生长在世界上难以获取的地区。植物合成生物碱的途径通常很长，有许多生物合成步骤。许多植物生物碱途径共有一些关键的中间化合物，而生物碱结构的复杂性来自于不同植物途径中不同的后续步骤。如果能够以更受控的方式制造一些关键的中间体生物碱化合物，并能够使用酶或化学来修饰这些生物碱结构，将是非常有益的。通过这种方式可以发现新的生物活性药物。几种生物碱的合成途径已经开始确定，但一些更复杂的生物碱的许多步骤仍然未知。在过去的几年中，研究人员已经开始分离一些生物碱合成中一些关键步骤的酶（及其基因）。其中一些酶执行非常重要的步骤，例如将源自简单氨基酸的相当简单的化合物连接在一起。我们打算利用全基因合成、合成生物学和途径设计技术来制造其中一些关键的生物碱合成酶，并将其放入实验室细菌中。通过这种方式，我们可以结合不同的植物酶和细菌酶，并建立合成途径来创建新的化合物，这些化合物可以筛选出有用的药物活性。通过细菌中的途径，我们将能够利用可再生资源以可重复的方式产生它们，而不依赖于困难或稀有植物的生长。此外，不需要使用目前用于合成此类异喹啉的方法进行广泛的化学转化，这些方法具有高环境影响。

项目成果

One-pot triangular chemoenzymatic cascades for the syntheses of chiral alkaloids from dopamine

• DOI: 10.1039/c4gc02325k
• 发表时间: 2015-01-01
• 期刊: GREEN CHEMISTRY
• 影响因子: 9.8
• 作者: Lichman, B. R.;Lamming, E. D.;Ward, J. M.
• 通讯作者: Ward, J. M.

The Catalytic Potential of Coptis japonica NCS2 Revealed - Development and Utilisation of a Fluorescamine-Based Assay

• DOI: 10.1002/adsc.201200641
• 发表时间: 2012-11-01
• 期刊: ADVANCED SYNTHESIS & CATALYSIS
• 影响因子: 5.4
• 作者: Pesnot, Thomas;Gershater, Markus C.;Hailes, Helen C.
• 通讯作者: Hailes, Helen C.

Library of Norcoclaurine Synthases and Their Immobilization for Biocatalytic Transformations.

去甲乌云碱合成酶文库及其用于生物催化转化的固定化。

• DOI: 10.1002/biot.201700542
• 发表时间: 2018
• 期刊: Biotechnology journal
• 影响因子: 4.7
• 作者: Lechner H

Tetrahydroisoquinolines affect the whole-cell phenotype of Mycobacterium tuberculosis by inhibiting the ATP-dependent MurE ligase.

• DOI: 10.1093/jac/dkv010
• 发表时间: 2015
• 期刊: The Journal of antimicrobial chemotherapy
• 作者: Guzman JD;Pesnot T;Barrera DA;Davies HM;McMahon E;Evangelopoulos D;Mortazavi PN;Munshi T;Maitra A;Lamming ED;Angell R;Gershater MC;Redmond JM;Needham D;Ward JM;Cuca LE;Hailes HC;Bhakta S
• 通讯作者: Bhakta S

'Dopamine-first' mechanism enables the rational engineering of the norcoclaurine synthase aldehyde activity profile.

• DOI: 10.1111/febs.13208
• 发表时间: 2015-03
• 期刊: The FEBS journal
• 作者: Lichman BR;Gershater MC;Lamming ED;Pesnot T;Sula A;Keep NH;Hailes HC;Ward JM
• 通讯作者: Ward JM