Investigating biosynthesis of the newly discovered natural product euglenatide and distribution across the breadth of Euglenoid algae

所有药物中有三分之一是基于天然产物，其中大多数目前是从细菌、真菌和植物中提取的。藻类是这些药物样分子的主要未开发来源，该研究计划将重点放在这些藻类上，以了解高价值化合物的生产。我们发现绿藻产生一种具有抗真菌和抗癌活性的天然产物，我们称之为绿藻内酯。我们想知道这些分子是如何形成的，它们的分布范围有多广，以及这些藻类还能制造出哪些潜在的新药。这个项目是由加拿大特伦特大学的一个研究小组和诺丁汉大学的一个研究小组合作完成的，前者是分离和基因工程类绿藻的专家，后者是绿藻天然产物鉴定的专家。微生物相互作用，无论是竞争还是合作，通过释放化学物质和许多涉及的天然产物已经被我们用作药物。这些药物包括一些最关键的抗生素，如青霉素和红霉素，以及抗癌药物，如阿霉素。藻类是一种多样化的生物，尽管有诱人的证据表明它们具有巨大的天然产物合成能力，但它们在很大程度上仍未得到研究。少数几种藻类会产生剧毒化合物，只有在有害藻类大量繁殖（如赤潮）导致人类和动物中毒时才会被发现。如果我们调整这种毒性，只对细菌或癌细胞起作用，我们就可以把毒素变成药物。有证据表明，许多其他类型的藻类可以产生多种生物活性化合物，这些化合物有可能被我们用作抗生素或抗癌剂。在这个研究项目中，我们要研究的藻类是绿藻，这是一组单细胞藻类，与绿藻有远亲关系。它们在世界各地的淡水池塘和沟渠中都有发现，其中一些品种被广泛用于研究和应用。绿藻能产生非常广泛的化合物，包括抗氧化剂、维生素和滋养大脑的多不饱和脂肪酸，甚至在废水中也能生长。这导致了Euglena作为一种食品补充剂的发展，以及正在进行的以商业规模生产碳中性生物燃料的工作。它们有能力在不适合农业的土地上种植，投入最少，产量明显高于传统作物，这给人们带来了巨大的希望。人们越来越有兴趣利用它们来生产高价值的蛋白质和药物。我们已经发现，Euglena基因组编码的基因可以产生广泛的天然产物，如果我们能够实现这一潜力，这将是非常有价值的。对Euglena的广泛学术和工业研究将有助于将该项目的成果迅速转化为社会效益。我们提出的研究计划将首先在一个经过充分研究的实验室菌株中使用基因工程来删除我们认为可能参与合成裸藻内酯的特定基因。然后，我们将看到哪些变化会阻止任何生产，哪些变化会产生其他产品。然后，我们将研究来自安大略省淡水湖的新的类绿藻菌株，看看这些藻类的野生菌株是否能产生类绿藻。我们还将研究在这些新培养的Euglena中其他分子的产生。我们的初步研究表明，这种类型的藻类具有很大的生物合成能力，可以制造有趣的分子，这项研究计划将使我们能够广泛地探索它们。通过在这些被研究的生物体中工作，我们最大限度地增加了发现以前从未见过的化合物的机会，然后将其作为生物活性探针和候选药物推进。独特的基因组编辑工具配对在真芽胞苷生产者给我们一个前所未有的机会来研究天然产物的生物合成途径。

One third of all pharmaceuticals are based on natural products, most of which are currently derived from bacteria, fungi and plants. Algae are a largely untapped source of these drug-like molecules and this research program will focus on these to understand the production of highly valuable compounds. We have found that the alga Euglena produces a natural product with antifungal and anticancer activity, which we called euglenatides. We want to find out how these molecules are made, how widespread they are and what other potential new drugs are made by these algae. This project is a collaboration between a group at Trent University in Canada who are experts in isolating and genome engineering Euglenoid algae, and a group at the University of Nottingham with expertise in Euglena natural product identification. Microbes interact, whether in competition or collaboration, through released chemicals and many of the natural products involved have been co-opted by us as pharmaceuticals. These include some of the most critical antibiotics, including penicillin and erythromycin, and cancer medicines, such as doxorubicin. Algae are a diverse group of organisms that, despite tantalising evidence for a vast capacity for natural product synthesis, have remained largely unstudied. A few species of algae produce highly toxic compounds, only discovered when harmful algal blooms, such as red tides, cause poisonings in humans and animals. If we tune this toxicity to only act against bacteria or cancer cells, we can turn toxins into medicines. There is evidence that many other types of algae make a wide range of bioactive compounds, which have the potential for us to use as antibiotics or anti-cancer agents. The algae we are going to work on in this research program are Euglena, a group of single-celled algae, which are distantly related to green algae. They are found in freshwater ponds and ditches, all over the world, and some species are used in a wide range of research and applications. Euglena produce an extraordinarily broad range of compounds, including antioxidants, vitamins and brain-nourishing polyunsaturated fatty acids, even grown in wastewater. This has led to the development of Euglena as a food supplement and ongoing work to produce carbon neutral biofuels at a commercial scale. There is great promise from their ability to grow on land unsuitable for agriculture, with minimal inputs, achieving significantly higher yields than from conventional crops, and there is a growing interest in engineering them to produce high value proteins and pharmaceuticals. We have found that the Euglena genome encodes genes for producing a wide range of natural products that will be highly valuable if we can realise this potential. The wide range of academic and industrial research into Euglena will help to rapidly translate the results of this project into societal benefits.The research plan we are proposing will initially use genetic engineering in a well-studied lab strain to delete specific genes we propose may be involved in the synthesis of the euglenatides. We will then see which changes prevent any production and which make other products. We will then look at new strains of Euglenoid algae from the freshwater lakes in Ontario to see if wild strain of these algae can make euglenatides. We will also investigate production of other molecules in these new cultures of Euglena. Our preliminary research suggests this type of algae has a large biosynthetic capacity to make interesting molecules and this research program will allow us to explore them extensively. By working in these under studied organisms we maximise our opportunities to find compounds that have never been seen before, which will then be advanced as bioactive probes and drug candidates.The unique pairing of genome editing tools in a euglenatide producer gives us an unprecedent chance to study the natural product biosynthetic pathway..