SBIR Phase I: Using enzymes secreted by soil amoebae to fight bacterial biofilms in agriculture

SBIR 第一阶段：利用土壤阿米巴原虫分泌的酶来对抗农业中的细菌生物膜

• 批准号: 2052039
• 负责人: Noah Stern
• 金额: $ 25.4万
• 依托单位: AGRAFILMS INC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2052039&HistoricalAwards=false
• 关键词: SBIR; Phase; Using; enzymes; secreted

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is development of a novel technology platform for the treatment of bacterial pathogens that pose a persistent threat to plant health both in the field and during storage of the harvest. Many bacteria kill their host by forming biofilms on the plant surface. Central to this proposal are findings that biofilms confer tolerance of bacteria to antimicrobial agents, whether these antimicrobial agents are antibiotics, disinfectants, or germicides. Dispersing these biofilms is necessary to remove contaminating bacteria from the plant surface. This proposal is addressing these challenges by developing safe and natural biofilm degrading enzymes from the collection of over 3,000 unique microorganisms called Dictyostelids isolated from five continents and several island nations. The focus of this proof-of-principle project is discovery of enzymes for treatment of bacterial potato soft rot. This disease is socially and commercially impacting our nation as potatoes are the most consumed vegetable in the U.S. with a total market value exceeding $3.5 B. The proposed project advances the use of Dictyostelid strains capable of dispersing biofilms formed by Dickeya and Pectobacterium and feeding on planktonic bacteria liberated from the corresponding bacterial biofilms. To be able to destroy those biofilms they must be able to break down biofilm integrity. This project uses proprietary technology licensed to the firm for a world-wide use in the agricultural markets. Suspected members of Glycoside Hydrolase Family of enzymes (GHF) have been already identified by bioinformatics and in crude secreted products. This project has two objectives: (1) Using polynucleotide chain reaction it will amplify Dictyostelids DNA encoding GHF and clone it into an Escherichia coli compatible vector with an in-frame Histidine-Tag sequence for purification using a specialized affinity column, and (2) Affinity purified enzymes will be used to disperse Dickeya and Pectobacterium biofilms established on 96-well polystyrene plates. The best performing enzymes will be further examined for their ability to disperse biofilms on a potato surface and prevent soft rot.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该小企业创新研究 (SBIR) 第一阶段项目的更广泛影响/商业潜力是开发一种新颖的技术平台，用于治疗细菌病原体，这些细菌病原体对田间和收成储存期间的植物健康构成持续威胁。许多细菌通过在植物表面形成生物膜来杀死宿主。该提案的核心是发现生物膜赋予细菌对抗菌剂的耐受性，无论这些抗菌剂是抗生素、消毒剂还是杀菌剂。分散这些生物膜对于去除植物表面的污染细菌是必要的。该提案通过从五大洲和几个岛国分离出的 3,000 多种独特微生物（称为盘基网柄菌）中开发安全、天然的生物膜降解酶来应对这些挑战。该原理验证项目的重点是发现用于治疗细菌性马铃薯软腐病的酶。 这种疾病对我们的国家产生了社会和商业影响，因为土豆是美国消费最多的蔬菜，总市场价值超过 3.5 美元。 B. 拟议项目推进使用能够分散由 Dickeya 和果杆菌形成的生物膜并以浮游生物为食的 Dictyostelid 菌株细菌从相应的细菌生物膜中释放出来。为了能够破坏这些生物膜，它们必须能够破坏生物膜的完整性。该项目使用了该公司授权的专有技术，可在全球农业市场上使用。生物信息学和粗分泌产物中已鉴定出糖苷水解酶家族 (GHF) 的可疑成员。该项目有两个目标：(1) 使用多核苷酸链式反应，扩增编码 GHF 的盘基网柄菌 DNA，并将其克隆到具有框内组氨酸标签序列的大肠杆菌兼容载体中，以便使用专门的亲和柱进行纯化，以及 (2)亲和纯化的酶将用于分散在 96 孔聚苯乙烯板上建立的 Dickeya 和果杆菌生物膜。将进一步检查性能最佳的酶在马铃薯表面分散生物膜和防止软腐病的能力。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。