Characterization and engineering of the biosynthesis of alternative painkilling natural products

植物界仍然是复杂的和生物活性的专门的代谢产物的药物的一个伟大的来源。开花植物提供了世界卫生组织列出的11%的基本药物。数百万年来，这些专门的代谢物一直在进化，同时它们参与植物-植物/微生物/动物通信和植物防御各种攻击。这种进化导致了它们巨大的结构多样性，从而导致了它们不同的生物活性。研究它们的生物化学组装以了解这种化学多样性的起源并通过新兴的合成生物学提供获得稀有生物活性化合物的解决方案是非常有趣的。作为新玩法大学的NBHRF/Tetra Bio-Pharma大麻健康研究主席，我研究大麻植物中专门代谢物大麻素生产的生物化学和分子调控。我的大麻研究计划是不可或缺的我的长期目标：i）了解如何从不同的植物物种的酶催化剂的演变有助于自然的化学多样性，阐明所涉及的生化途径，和ii）识别和利用这些酶制造重要的天然产品。NSERC的支持将有助于实现这一目标。在资助期间，我将专注于替代止痛天然产品的生物合成的表征和工程，这些产品可能取代有效但有问题的阿片类药物。短期目标包括阐明1）conolidine和2）perrottetinene的完整生物合成，以及3）工程化其微生物生产用于工业制造。与阿片类药物不同的是，康诺利定是一种强效但非成瘾性的镇痛药。然而，其在来源植物中的含量极低以及化学合成中存在的挑战阻碍了其作为药物的发展。研究conolidine的生物合成也有助于了解一个重要的代谢物类的化学进化：单萜吲哚生物碱，具有令人印象深刻的结构复杂性和多样性，使他们迷人的生物活性。 分离自齿舌属（Radula spp.）在地钱门中，perrottetinene显示出与来自维管植物大麻（Cannabissativa）的四氢大麻酚（THC）惊人的结构相似性，大麻被认为是大麻素的唯一物种，并且在4.8亿年前从地钱中分化出来。由于内源性大麻素系统已成为疼痛管理的靶点，作为人类大麻素受体配体，perrottetinene可能被开发成一种有前途的镇痛药。通过将这种物质与THC的生物合成进行比较，我们的目标是回答这些结构相似的化合物是如何在两个远亲物种中进化的，并开发用于生产大麻素镇痛剂的新酶催化剂。

The plant kingdom remains a great source for complex and bioactive specialized metabolites for medicine. Flowering plants provide 11% of the essential medicines listed by the World Health Organization. For millions of years, these specialized metabolites have been evolving while they participate in plant-plant/microbe/animal communications and plant defense against various attacks. This evolution has resulted in their great structural diversity responsible for their diverse bioactivities. It is of great interest to study their biochemical assembly to understand the origin of such chemo-diversity and to provide solutions to access the rare bioactive compounds by emerging synthetic biology. As the NBHRF/Tetra Bio-Pharma Cannabis Health Research Chair at University of New Brunswick, I study the biochemistry and molecular regulations of the production of specialized metabolites, cannabinoids, in the cannabis plant. My cannabis research program is integral to my long-term objectives: i) to understand how the evolution of enzyme catalysts from different plant species contributes to Nature's chemo-diversity by elucidating the biochemical pathways involved, and ii) to identify and harness these enzymes for manufacturing important natural products. Support from NSERC will contribute to achieving this ambition. During the grant period, I will focus on the characterization and engineering of the biosynthesis of alternative painkilling natural products that may replace effective but problematic opioids. Short-term objectives include elucidating the complete biosynthesis of 1) conolidine, and 2) perrottetinene, and 3) engineering their microbial production for industrial manufacturing. Operating in a mode different from opioids, conolidine is a potent but non-addictive analgesic. Its extremely low content in source plants and the challenges remaining in chemical synthesis, however, impede its development as a medicine. Studying the biosynthesis of conolidine also contributes to understanding the chemical evolution of an important metabolite class: monoterpenoid indole alkaloids, which have impressive structural complexity and diversity that gives them fascinating bioactivities. Isolated from Radula spp. in the phylum of liverwort, perrottetinene shows astonishing structural similarity to tetrahydrocannabinol (THC) from the vascular plant Cannabis sativa, which was known as the only species for cannabinoids and diverged from liverworts 480 million years ago. As a human cannabinoid receptor ligand, perrottetinene may be developed into a promising analgesic, since the endocannabinoid system has become a target in pain management. By comparing this substance to the biosynthesis of THC, we aim to answer how these structurally similar compounds have evolved in two distantly related species, and also develop new enzyme catalysts for producing cannabinoid analgesics.