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Fusion protein-based biopesticides for sustainable crop protection

用于可持续作物保护的融合蛋白生物农药

基本信息

批准号：
TS/I000690/1
负责人：
John Gatehouse
金额：
$ 7.33万
依托单位：
Durham University
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2011
资助国家：
英国
起止时间：
2011 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=TS%2FI000690%2F1
关键词：
Fusion protein based biopesticides sustainable

项目摘要

This research and development programme is intended to result in the introduction of a novel, environmentally beneficial biopesticide to market, as a replacement for older pesticides being removed from use under legislation on pesticide safety introduced by the EC. The biopesticide is based on fusion protein technology invented and developed collaboratively by the Food and Environment Research Agency (Fera), York and Durham University. This allows selected toxins from arthropods, which have no toxicity towards higher animals, to be combined with a carrier protein that makes them orally toxic to invertebrates, whereas they would normally only be effective when injected into a prey organism by a predator. The fusion protein, containing both the toxin and the carrier is produced as a recombinant protein in a microbial expression system, which can be scaled up for industrial production.Research previously funded under LINK programmes has resulted in a best candidate fusion protein ( FP4 ) which has been evaluated for commercial production. However, in the present programme it is intended to use a 2010 best candidate fusion protein with improved insecticidal activity ( FP5 ), which will be more likely to result in a commercially viable product. In order to bring this technology to market, the laboratory scale process for protein production must be optimised, to decrease production costs. Yield of recombinant protein per volume of microbial culture is a major factor in the final cost of product, and therefore the initial stage of the programme will be to improve the existing expression clones. Experience with an earlier fusion protein has shown that improvements in yield of product can be made by the introduction of extra copies of gene sequence into the yeast expression host, and this will be carried out for the 2010 best candidate . The resulting yeast clones will be screened for protein production, and the best clones will be selected and taken forward in the programme. The aim will be to produce recombinant protein on a 1g per litre of microbial culture scale.If a fusion protein-based biopesticide is to be introduced as a commercial product, testing will be necessary to assess its effects on non-target organisms. The toxin selected for this programme has been shown to be non-toxic to higher animals. However, further testing of the fusion protein will need to be carried out by other partners in the programme. A more significant potential problem is effects on beneficial invertebrates, such as the predators and parasites that attack crop pests, pollinators such as bees, and earthworms. Earlier experiments with the best candidate fusion protein, carried out under a previously funded LINK programme, have suggested that fusion proteins do not cause adverse effects on beneficial organisms. However, more extensive testing of the 2010 best candidate fusion protein will be required to confirm absence of any adverse environmental impact. The academic partners will carry out preliminary work in this area, using a service provided by Newcastle University, prior to more extensive testing required to meet regulatory requirements, which will be addressed by industrial partners.Finally, further development of fusion proteins will be carried out by Durham and Fera, separately from the present programme. If fusion proteins with higher insecticidal activity or other desirable properties are developed, these will be made available for development by industrial partners in the TSB programme.

这项研究和开发计划的目的是向市场推出一种新的、对环境有益的生物农药，以取代根据欧洲共同体提出的农药安全立法而被淘汰的旧农药。这种生物杀虫剂是基于融合蛋白技术，由食品和环境研究局（Fera）、约克和达勒姆大学合作发明和开发。这使得来自节肢动物的选定毒素（对高等动物没有毒性）与载体蛋白结合，使其对无脊椎动物具有口服毒性，而它们通常只有在被捕食者注射到猎物生物体时才有效。这种融合蛋白含有毒素和载体，在微生物表达系统中以重组蛋白的形式生产，可以扩大规模用于工业生产。先前在LINK项目下资助的研究已经产生了一种最佳候选融合蛋白（FP4），该蛋白已被评估用于商业生产。然而，在本方案中，打算使用具有改进的杀虫活性的2010年最佳候选融合蛋白（FP5），这将更有可能产生商业上可行的产品。为了将这项技术推向市场，必须优化蛋白质生产的实验室规模工艺，以降低生产成本。每单位体积微生物培养物的重组蛋白产量是产品最终成本的主要因素，因此该计划的初始阶段将是改进现有的表达克隆。早期融合蛋白的经验表明，可以通过将额外的基因序列拷贝引入酵母表达宿主中来提高产物的产量，这将在2010年最佳候选者中进行。将对产生的酵母克隆进行蛋白质生产筛选，并选择最佳克隆并在计划中继续进行。目标是以每升微生物培养物1克的规模生产重组蛋白，如果以融合蛋白为基础的生物农药作为商业产品推出，将需要进行测试，以评估其对非目标生物的影响。为该方案选择的毒素已被证明对高等动物无毒。然而，融合蛋白的进一步测试将需要由该方案的其他合作伙伴进行。一个更重要的潜在问题是对有益的无脊椎动物的影响，例如攻击作物害虫的捕食者和寄生虫，蜜蜂和蚯蚓等授粉者。在先前资助的LINK计划下进行的最佳候选融合蛋白的早期实验表明，融合蛋白不会对有益生物产生不利影响。然而，需要对2010年最佳候选融合蛋白进行更广泛的测试，以确认没有任何不利的环境影响。学术合作伙伴将利用纽卡斯尔大学提供的服务在这一领域开展初步工作，然后进行更广泛的测试，以满足监管要求，这将由工业合作伙伴解决。最后，融合蛋白的进一步开发将由达勒姆和Fera独立于目前的计划进行。如果开发出具有更高杀虫活性或其他所需特性的融合蛋白，TSB计划的工业伙伴将提供这些蛋白供开发。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Novel biopesticide based on a spider venom peptide shows no adverse effects on honeybees.

DOI：
10.1098/rspb.2014.0619
发表时间：
2014-07-22
期刊：
Proceedings. Biological sciences
影响因子：
0
作者：
Nakasu EY;Williamson SM;Edwards MG;Fitches EC;Gatehouse JA;Wright GA;Gatehouse AM
通讯作者：
Gatehouse AM

A recombinant fusion protein containing a spider toxin specific for the insect voltage-gated sodium ion channel shows oral toxicity towards insects of different orders.