PFI:AIR - TT: Using A. niger as a host to express recombinant endolysin

这个PFI：空气技术翻译项目关注的是一个与农业和人类健康相关的重要问题。50多年来，在动物饲料中添加低剂量的传统(即“医学上重要的”)抗生素，以提高食用动物的生产效率，一直是农业行业的普遍做法。虽然这些“抗生素生长促进剂”提供了巨大的经济效益，但这种做法也导致了具有抗生素耐药性的细菌菌株，有可能感染人类。因此，美国食品和药物管理局已经采取措施阻止这种做法。虽然已经提出了许多具有重要医学意义的抗生素的替代品，但大多数都没有在商业上成功，因为它们不够有效。作为医学上重要抗生素的替代品，抗菌蛋白的使用前景广阔，但其商业化生产的一个重大限制是成本。因此，经济地制造抗菌蛋白的能力将对农业产生重大好处，因为这将使农民能够保持高水平的生产力，同时不会产生对人类构成危险的抗药性菌株。这项建议探索了用抗菌蛋白(AMP)取代具有重要医学意义的抗生素饲料添加剂的可能性。这里描述的工作试图使用重组丝状真菌产生概念验证数据，以商业上可行的滴度表达模型AMP。该项目使用商业上相关的真菌物种(黑曲霉)来表达具有杀菌活性的模型内溶蛋白。为了增加两种不同的内溶素酶的表达和细胞外分泌，将使用几种方法，探索基因组位置、转录启动子和分泌信号序列的影响。采用几种方法将增加成功的可能性，并将有助于确定导致高产重组真菌菌株的最相关因素。这一理解将允许有效地为不同的蛋白质或多肽产生额外的生产菌株。此外，参与该项目的人员，包括一名博士后科学家和几名本科生，将在跨学科环境中接受培训，以提高他们的研究能力。人员还将通过开展与该项目相关的客户发现活动，参与创业教育体验。这将主要涉及在整个价值链上联系潜在的利益相关者，以测试与未来商业化相关的假设。

This PFI:AIR Technology Translation project focuses on an important agricultural and human-health related problem. For over 50 years, it is has been common practice in the agricultural industry to add low doses of traditional (i.e., "medically important") antibiotics to animal feed to improve production efficiency of food animals. While these "antibiotic growth promoters" provide tremendous economic benefit, this practice also leads to antibiotic-resistant bacterial strains which have the potential to infect humans. As a result, the US Food and Drug Administration has implemented measures to stop this practice. While numerous alternatives to medically important antibiotics have been proposed, most have not been commercially successful as they are not adequately effective. As an alternative to medically important antibiotics, the use of antimicrobial proteins has great promise, but a significant limitation to their commercial production has been cost. Thus the ability to economically manufacture antimicrobial proteins would represent a significant benefit to the agricultural industry as it would allow farmers to maintain high levels of productivity while not generating antibiotic-resistant strains that pose a danger to humans. This proposal explores the potential of replacing medically-important antibiotic feed-additives with antimicrobial protein (AMP). Work described here seeks to generate proof-of-concept data, expressing model AMPs, at commercially viable titers, using recombinant filamentous fungi.This project employs a commercially-relevant fungal species (Aspergillus niger), to express model endolysin proteins with demonstrated bactericidal activity. To increase both expression and extracellular secretion of two different endolysin enzymes several approaches will be used, exploring the impact of genomic location, transcriptional promoter, and secretion signal sequence. Employing several approaches will increase the likelihood for success and will also allow identification of the most relevant factors leading to a high-producing recombinant fungal strain. This understanding will allow efficient generation of additional production strains for different proteins or peptides. In addition, personnel involved in this project, both a postdoctoral scientist and several undergraduate students, will be trained in a cross-disciplinary environment that will increase their research capacity. Personnel will also participate in entrepreneurial education experiences, through conducting customer discovery activities related to this project. This will primarily involve contacting potential stakeholders, along the entire value chain, to test hypotheses related to future commercialization.