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

合作研究：工程形态和蛋白质分泌以提高丝状真菌发酵的生产力

• 批准号: 1159933
• 负责人: Steven Harris
• 金额: $ 27.49万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1159933&HistoricalAwards=false
• 关键词: Collaborative; Research; Engineering; morphology; protein

1159973/1159933 Marten/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.

1159973/1159933 Marten/Harris丝状真菌能够在致晕中产生和分泌蛋白质（即， 100 g/L）。然而，对于许多重组蛋白，生产率低得多，通常是由于蛋白分泌不足。该项目的总体目标是更深入地了解丝状真菌中的蛋白质分泌，以便制定策略来提高真菌发酵过程中的生产力。假设真菌蛋白质分泌和菌丝形态密切相关，因此形态突变体的鉴定也将选择低分泌型和高分泌型。为了检验这一假设，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表征这些突变体（即，基因组测序和蛋白质组学分析）以鉴定负责其表型的基因和分子机制。这种经典遗传筛选与现代“组学”技术的结合将允许快速鉴定基因功能。丝状真菌广泛用于生物加工工业，生产价值数十亿美元的大量化合物。相比之下，真菌病原体是一个重要的问题，导致人类的显著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可以提高真菌生物过程的生产力，而且有可能确定改进抗真菌疗法的新靶点。所产生的Ts突变体库将提供给更广泛的真菌研究界，并具有潜在的转化影响，因为真菌形态和蛋白质分泌在几乎所有丝状真菌中起着重要作用。这项研究还将对人力资本产生广泛影响，因为它具有重要的教育组成部分。大部分突变体筛选工作将由本科生完成，这为许多人提供了第一次接触研究的机会。在研究生阶段，学生将在分子生物学和工程的接口进行培训，从而获得宝贵的跨学科经验。他们还将获得监督经验，因为他们帮助协调参与该项目的本科生的研究。