Engineering yeast membranes with mimetic and custom architecture for improved performance in biotechnological applications

酵母是现代生物技术的主力，因其健壮性和多功能性而受到工业的青睐。如今，从药物到小分子、酶和疫苗、人母乳成分、肉类替代品血红素、生物塑料、生物材料甚至生物燃料等各种产品都是在酵母中生产的。尽管人们在了解和设计高效的酵母细胞工厂方面做了大量的努力，但对酵母膜特性及其对生物技术应用的贡献的研究却很少。几个过程，无论是生理的还是工程的，都与膜有很大的关系，并且可能限制酵母细胞工厂的性能。目前的研究方法主要是将生物膜作为异源宿主产生的化合物毒性作用的主要靶点。相反，对于宿主微生物与天然膜的膜组成差异如何影响不同来源的生物合成酶的功能能力，以及如何干扰代谢物的细胞内运输及其向细胞外的输出，我们知之甚少，也无法预测。拟议研究的长期目标是设计下一代生物技术应用的生产平台，考虑到生物膜的作用。在短期内，我们将直接解决与膜在异源酶的功能和途径中的作用有关的方面，通过修改酵母膜的组成来模仿生产生物或通过工程改造非天然酵母膜。因此，我们将通过设计仿生植物膜或定制酵母来开发植物酵母，并将脂滴重新用于目标化合物的储存室。利用这些平台，我们将评估模型萜类生物合成途径的性能和代谢复合物的形成。这个项目将有利于科研团体和加拿大的工业。我们致力于开发先进的酵母底盘，以获得具有有趣生物活性的高价值分子，并为化学生产提供可持续的解决方案。因此，这些化合物可以更广泛地用于医药、农业和其他工业应用。在这里开发的知识将提供有效的方法来研究和工程生物膜和相关过程的领域。它还将促进工业创新，并将解决加拿大商业研发和制造活动的优先事项。能够加快上市时间和增加经济收益的改进生产平台将极大地有利于加拿大经济。因此，这一研究项目在未来几年的相关性跨越了多个层面，包括推进科学发现，解决当前和未来的工业挑战，以及为新战略和创新建立强大的基础。

Yeast is a workhorse of modern biotechnology, preferred by industry due to its robustness and versatility. A broad range of products from medicines to small molecules, enzymes and vaccines, components of human breast milk, heme for meat substitutes, bioplastics, biomaterials and even biofuels are nowadays produced in yeast. Despite extensive efforts to understand and engineer efficient yeast cell factories, less attention has been placed on studying yeast membrane properties and their contribution to biotechnological applications. Several processes, both physiological or engineered, are greatly related to membranes, and could be limiting the performance of yeast cell factories. Current approaches mainly address the role of biological membranes as the primary target of the toxic effects of compounds produced in heterologous hosts. In reverse, there is little understanding of, or ability to predict, how differences in membrane composition of the host microorganism versus native membranes affect functional capacity of biosynthetic enzymes of different origin and interfere with intracellular transport of metabolites and their export outside of the cell. The long-term goal of the proposed research is to engineer the next generation of production platforms for biotechnological applications taking in consideration the role of biological membranes. In the short-term, we will directly address aspects related to membrane role in functionality of heterologous enzymes and pathways by modifying the yeast membrane composition to mimic that of producer organisms or by engineering non-natural yeast membranes. Thus, we will develop Phyto-yeast by biomimicking plant membrane or tailored yeast by design, and repurpose lipid droplets into storage compartments of target compounds. Using these platforms, we will evaluate the performance of model terpenoid biosynthetic pathways and formation of metabolic complexes. This program will benefit both the research scientific community and Canadian industry. We endeavor to develop advanced yeast chasses that will enable accessing high-value molecules with interesting biological activities and offer sustainable solutions to chemical production. Consequently, these compounds can be more broadly exploited for medicinal, agricultural, and other industrial applications. The knowledge developed here will provide the field with effective approaches to study and engineer biological membranes and the related processes. It will also foster innovations in industry and will address Canadian priorities for commercial R&D and manufacturing activities. Improved production platforms capable of accelerating time to market and increasing economic gain will highly benefit the Canadian economy. Thus, the relevance of this research program for the years ahead span multiple levels, including advancing scientific discoveries, tackling current and future industrial challenges, and building strong underpinning for novel strategies and innovations.