An integrated approach to camptothecin biosynthesis discovery and engineering

植物生物活性化合物研究实验室（PlantBioCore）旨在解码有助于植物专业代谢的巨大多样性的基因组和生物化学特征，并将这些知识转化为创新的生物技术，以满足高价值化学品的社会需求。 植物产生超过200，000种不同的特殊代谢物，其中许多是宝贵的营养素，商品和治疗药物。它们的结构复杂性表明它们的生物合成途径是合成生物学应用的新型生物催化剂的杰出来源。不幸的是，在植物专门代谢的研究进展一直滞后，在很大程度上是由于植物系统的复杂性。喜树碱（抗癌）、紫杉醇（抗癌）、奎宁（抗疟疾）只是尚未表征的重要植物代谢途径的几个例子，因此开发这些丰富的代谢资源的方法尚未实现。 PlantBioCore的直接重点是抗癌化合物喜树碱，由喜树树产生。我们将整合一套创新的现代生物信息学，分析，生物化学和分子遗传学方法，以解开喜树碱和相关有价值的植物化学物质的生物合成中前所未有的生物化学。短期目标是：（i）发现新的生物合成酶;（ii）在功能和结构上表征这些生物催化剂;（iii）产生生物碱结构和功能二聚体;和（iv）重建喜树碱生物合成途径的部分。对生物合成中酶促步骤的初步了解将为我们对这些酶和代谢产物的时空组织的长期研究提供信息，为在5-10年内成功改造和重建喜树碱以及其他生物碱途径铺平道路。对喜树碱生产的全面了解将使我们能够获得植物特有的化学转化，指导新的生物代谢途径的发现，阐明现有途径的演变，并为代谢工程建立一个有利的酶工具箱。 除了生物碱生物合成的更广泛和详细的图片，我们的研究计划为高素质的人才提供了独特的机会，以发展生物信息学，酶学，生物化学，项目管理和科学交流的先进技术技能。这些经验将使他们在以研究为导向的就业市场上具有很强的竞争力，并有助于加拿大知识驱动型经济的增长。我们的研究计划将阐明大自然如何从几种简单的起始材料中创造出广泛的结构多样性，并为从植物中提取化学物质或从石化产品中合成提供可持续的替代方案，从而使加拿大学术界和公共部门受益。

The Plant Bioactive Compounds Research (PlantBioCoRe) Laboratory aims to decode the genomic and biochemical features that contribute to the vast diversity of plants' specialized metabolism, and to translate this knowledge into innovative biotechnologies to meet the societal needs of high-value chemicals. Plants produce upwards of 200,000 distinct specialized metabolites, many of which are invaluable nutrients, commodity products, and therapeutics. Their structural complexity signifies that their biosynthetic pathways are outstanding sources of new biocatalysts for synthetic biology applications. Unfortunately, research progress in plant specialized metabolism has lagged, in large part due to the complexities of plant systems. Camptothecin (anticancer), Taxol (anticancer), quinine (antimalarial) are just a few examples of the important plant metabolic pathways that remain uncharacterized, and approaches to exploit these rich metabolism resources have thus been unrealized. PlantBioCoRe's immediate focus is on the anticancer compound campothecin, produced by the tree Camptotheca acuminata. We will integrate a suite of innovative and modern bioinformatic, analytical, biochemical, and molecular genetic approaches to unravel the unprecedented biochemistry harboured within the biosynthesis of camptothecin and related valuable phytochemicals. The short-term objectives are: (i) to discover new biosynthetic enzymes; (ii) to functionally and structurally characterize these biocatalysts; (iii) to generate alkaloid structural and functional diversities; and (iv) to reconstitute parts of the camptothecin biosynthetic pathway. Initial understanding of enzymatic steps in the biosynthesis will inform our long-term investigation on the temporal and spatial organization of these enzymes and metabolites, paving the way for successful engineering and reconstitution of not only camptothecin but also other alkaloid pathways in 5-10 years. A comprehensive understanding of camptothecin production will allow us to provide access to chemical transformations unique to plants, guide the discovery of new biosynthtic pathways, illuminate the evolution of existing pathways, and build an enabling enzymatic toolbox for metabolic engineering. In addition to a more expanded and detailed picture of alkaloid biosynthesis, our research program provides unique opportunities for highly-qualified personnel to develop advanced technical skills in bioinformatics, enzymology, biochemistry, project management and scientific communication. These experiences would allow them to be highly competitive in the research-oriented job market and contribute to the growth of Canada's knowledge-driven economy. Our research program will illuminate how Nature creates extensive structural diversity from a few simple starting materials, and offer sustainable alternatives to chemical extraction from plants or synthesis from petrochemicals, therefore, benefiting both Canadian academic and public sectors.