Protein engineering to enhance the commercial potential of novel fusion protein based biopesticides.

由于化学农药从市场上撤出以及新活性物质的短缺，伴随着对目前允许使用的少数产品的抗性的广泛发展，作物保护已经变得越来越成问题。申请人已经为一种方法申请了专利，该方法将缺乏口服毒性的天然来源的肽（例如蜘蛛捕获猎物时产生的麻痹毒素）转化为口服有效的生物杀虫剂。无脊椎动物特异性毒素与能够穿过肠壁的“载体”蛋白融合，并且融合蛋白的摄取使得毒素能够经由循环系统转运至目标害虫的中枢神经系统。在酵母中产生的重组原型融合蛋白对一系列作物害虫有效，包括蛞蝓、甲虫和蚜虫，但对蜜蜂没有有害影响。目前正在与工业伙伴合作，通过外源应用（如毒饵和喷雾剂）对这一技术进行商业开发，以控制作物害虫。随着表征蜘蛛毒液毒素的研究的进展，靶向不同电压门控离子通道以用于产生融合蛋白的可用无脊椎动物特异性肽毒素的库正在增加。现有的、完善的原型融合蛋白含有与载体融合的单一毒素，但申请人的新的可专利研究已经证明，可以通过蛋白质工程进一步增强功效。该项目旨在探索利用最新成果提高疗效的机会，从而提高这项新技术的商业化前景。宿主实验室在重组融合蛋白的设计和生产方面拥有丰富的经验，这些融合蛋白将构成学生进行研究计划的基础。无脊椎动物生理学方面的专业知识将进一步支持研究，以评估新型融合蛋白对作物害虫的功效和作用方式。该项目得益于与工业合作伙伴建立的合作关系。CASE合作伙伴的投入对于正在进行的合作研发至关重要，以开发生物农药融合蛋白作为平台技术。来自非学术合作伙伴的投入将确保拟议的研究项目重点和产出有最好的机会被成功翻译，以在商业环境中采用。

Crop protection has become increasingly problematic due to the withdrawal of chemical pesticides from the marketplace and a shortage of new actives concomitant with the widespread development of resistance to the few products currently permitted for use. The applicants have patented an approach that transforms naturally derived peptides that lack oral toxicity, such as paralytic toxins produced by spiders to capture prey, into orally effective biopesticides. Invertebrate specific toxins are fused to a "carrier" protein able to cross the gut wall and ingestion of the fusion protein enables transport of the toxin to the central nervous system of the target pest via the circulatory system. Recombinant prototype fusion proteins produced in yeast are effective against a range of crop pests, including slugs, beetles and aphids, but have no deleterious effects on bees. Commercial exploitation of this technology through exogenous applications (e.g. baits and sprays) for the control of crop pests is being developed in collaboration with the industrial partner. The pool of available invertebrate specific peptide toxins that target different voltage gated ion channels for use in the generation of fusion proteins is increasing as research to characterize spider venom toxins advances. The existing, well established prototype fusion proteins contain a single toxin fused to the carrier but new patentable research by the applicants has demonstrated that efficacy can be further enhanced through protein engineering. The project seeks to explore opportunities to build upon recent results to enhance efficacy and thereby commercialisation prospects for this novel technology. The host laboratory has extensive experience in the design and production of recombinant fusion proteins that will form the basis of the research programme to be conducted by the student. Expertise in invertebrate physiology will further support research to evaluate the efficacy and mode of action of novel fusion proteins against crop pests. This project benefits from an established collaborative relationship with the industrial partner. The input of the CASE partner is essential for ongoing collaborative R & D to develop biopesticidal fusion proteins as a platform technology. Input from the non-academic partner will ensure that the proposed research project focus and outputs have the best chance of being successfully translated for adoption in a commercial setting.