Improving the binding specificity and affinity of prenyltransferase enzymes using computational protein design********

四氢大麻酚(THC)和大麻二酚(CBD)等大麻素正成为治疗各种人类疾病的重要药物，如慢性疼痛、癫痫和帕金森氏病。目前，这些化合物是从大麻植物中提取的，大麻除了THC和CBD外，还会产生数百种活性生物化合物。这些额外化合物的存在使得将紫花苜蓿提取物标准化为药物变得困难。为了解决这个问题，Hyasynth Bio正在开发一种基于酵母发酵的高纯度大麻类化合物的替代生产方法。这种方法依赖于包括非植物戊烯基转移酶在内的异源生物合成途径，该途径催化从相应的异戊二烯基二磷酸代谢物衍生的C5(二甲基烯丙基)、C10(香叶基)或C15(法尼基)戊烯基转移到各种芳香受体上。然而，这些戊烯基转移酶表现出广泛的底物和区域特异性，使它们能够与许多不需要的芳香酵母代谢物反应，或者在错误的位置上合成所需的芳香酵母代谢物，这降低了该途径的整体效率。为了提高发酵过程的效率，从而提高这些酶对制药和娱乐大麻市场的价值，我们将使用计算蛋白质设计来设计具有窄特异性和增强与其目标底物结合亲和力的异丙苯基转移酶变体。这些经过改造的异丙烯基转移酶将直接整合到Hyasynth Bio的代谢工程和发酵工艺管道中，从而为该公司在大麻行业提供竞争优势，并直接为加拿大生物经济的发展做出贡献。**

Cannabinoids such as tetrahydrocannabinol (THC) and cannabidiol (CBD) are becoming increasingly important pharmaceuticals for the treatment of a variety of human diseases such as chronic pain, epilepsy, and Parkinson's disease. Currently, these compounds are extracted from the plant Cannabis sativa, which produces hundreds of active biological compounds in addition to THC and CBD. The presence of these additional compounds makes it difficult to standardize C. sativa extracts to be used as medicine. To address this issue, Hyasynth Bio is developing an alternative production method for high purity cannabinoids that is based on yeast fermentation. This method relies on a heterologous biosynthetic pathway including non-plant prenyltransferases, which catalyze the transfer of a C5 (dimethylallyl), C10 (geranyl) or C15 (farnesyl) prenyl group derived from the corresponding isoprenyl diphosphate metabolites onto a variety of aromatic acceptors. However, these prenyltransferases display broad substrate and regio-specificity, making them able to react with many undesired aromatic yeast metabolites or prenylate desired ones at incorrect positions, which lowers the overall efficiency of the pathway. To enhance the efficiency of the fermentation process and thereby the value of these enzymes to the pharmaceutical and recreational cannabis markets, we will use computational protein design to engineer prenyltransferase variants displaying narrow specificity and enhanced binding affinity to their target substrates. These engineered prenyltransferases will directly integrate into Hyasynth Bio's metabolic engineering and fermentation process pipelines, thereby providing the company with a competitive edge in the cannabis industry and directly contributing to the development of the Canadian bio-economy.**