Collaborative Research: Engineering morphology and protein secretion to enhance productivity in filamentous fungal fermentations

marten /Harris丝状真菌能够以惊人的数量（即100 g/L）生产和分泌蛋白质。然而，许多重组蛋白的生产效率要低得多，通常是由于蛋白质分泌不足。该项目的总体目标是对丝状真菌的蛋白质分泌有更深入的了解，以便制定策略来提高真菌发酵过程中的生产力。据推测，真菌蛋白分泌和菌丝形态密切相关，因此形态突变的鉴定也将选择低分泌和高分泌。为了验证这一假设，将开发具有异常蛋白质分泌的温度敏感（Ts）形态突变体。然后将使用现代工具（即基因组测序和蛋白质组学分析）对这些突变体进行表征，以确定导致其表型的基因和分子机制。这种经典基因筛选与现代“组学”技术的结合将使基因功能的快速鉴定成为可能。丝状真菌在生物加工工业中被广泛使用，生产出一系列价值数十亿美元的化合物。相比之下，真菌病原体是一个重大问题，造成人类大量发病率和死亡率，以及每年数十亿美元的作物损失。从这项工作中获得的见解不仅将提高真菌生物过程的生产率，而且有可能确定改进抗真菌治疗的新靶点。由于真菌形态和蛋白质分泌在几乎所有丝状真菌中都起着重要作用，因此所产生的Ts突变文库将提供给更广泛的真菌研究界，并具有潜在的变革性影响。这项研究也将对人力资本产生广泛的影响，因为它具有重要的教育成分。许多突变体筛选工作将由本科生完成，提供许多第一次接触研究。在研究生阶段，学生将在分子生物学和工程学的界面上接受训练，从而获得宝贵的跨学科经验。他们还将获得管理经验，因为他们帮助协调参与该项目的本科生的研究。

1159973/1159933Marten/Harris Filamentous fungi are capable of producing and secreting proteins in stunning (i.e., 100 g/L) quantities. Yet for many recombinant proteins productivity is much lower, often due to inadequate protein secretion. The overarching goal in this project is to develop a deeper understanding of protein secretion in filamentous fungi, so that strategies can be developed to improve productivity during fungal fermentation. It is hypothesized that fungal protein secretion and hyphal morphology are intimately linked, such that identification of morphological mutants will also select for both hypo- and hyper-secretors. To test this hypothesis, temperature sensitive (Ts) morphological mutants with aberrant protein secretion will be developed. These mutants will then be characterized using modern tools (i.e., genome sequencing and proteomic analysis) to identify the genes and molecular mechanisms responsible for their phenotypes. This combination of a classical genetic screen with modern "-omics" technology will allow rapid identification of gene function.Filamentous fungi are used extensively in the bioprocess industry to produce a vast array of compounds worth billions of dollars. In contrast, fungal pathogens are a significant problem, responsible for significant morbidity and mortality in humans as well as billions of dollars in crop losses annually. Insight gained from this work, will not only improve productivity in fungal bioprocesses but has the potential to identify novel targets for improved antifungal therapies. The Ts mutant library generated will be made available to the broader fungal research community and has the potential for transformational impact because of the important role fungal morphology and protein secretion play in nearly all filamentous fungi. This research will also have broad impact on human capital, as it has a significant educational component. Much of the mutant screening work will be accomplished by undergraduate students, providing many a first exposure to research. At the graduate level, students will be trained at the interface of molecular biology and engineering and thus will gain valuable interdisciplinary experience. They will also gain supervisory experience as they help coordinate the research of undergraduates participating in the project.