Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China.
J Nanobiotechnology. 2022 Jul 26;20(1):347. doi: 10.1186/s12951-022-01557-9.
The disease caused by plant pathogenic bacteria in the production, transportation, and storage of many crops has brought huge losses to agricultural production. N-acylhomoserine lactonases (AHLases) can quench quorum-sensing (QS) by hydrolyzing acylhomoserine lactones (AHLs), which makes them the promising candidates for controlling infections of QS-dependent pathogenic bacteria. Although many AHLases have been isolated and considered as a potentially effective preventive and therapeutic agents for bacterial diseases, the intrinsically poor ambient stability has seriously restricted its application.
Herein, we showed that a spheroid enzyme-based hybrid nanoflower (EHNF), AhlX@Ni(PO), can be easily synthesized, and it exhibited 10 times AHL (3OC8-HSL) degradation activity than that with free AhlX (a thermostable AHL lactonase). In addition, it showed intriguing stability even at the working concentration, and retained ~ 100% activity after incubation at room temperature (25 °C) for 40 days and approximately 80% activity after incubation at 60 °C for 48 h. Furthermore, it exhibited better organic solvent tolerance and long-term stability in a complicated ecological environment than that of AhlX. To reduce the cost and streamline production processes, CSA@Ni(PO), which was assembled from the crude supernatants of AhlX and Ni(PO), was synthesized. Both AhlX@Ni(PO) and CSA@Ni(PO) efficiently attenuated pathogenic bacterial infection.
In this study, we have developed N-acylhomoserine lactonase-based hybrid nanoflowers as a novel and efficient biocontrol reagent with significant control effect, outstanding environmental adaptability and tolerance. It was expected to overcome the bottlenecks of poor stability and limited environmental tolerance that have existed for over two decades and pioneered the practical application of EHNFs in the field of biological control.
许多作物在生产、运输和储存过程中,植物病原菌引发的疾病给农业生产带来了巨大损失。N-酰基高丝氨酸内酯酶(AHLases)可以通过水解酰基高丝氨酸内酯(AHLs)来淬灭群体感应(QS),这使它们成为控制依赖 QS 的病原菌感染的有前途的候选物。尽管已经分离出许多 AHLases,并被认为是预防和治疗细菌疾病的潜在有效药物,但内在的环境稳定性差严重限制了其应用。
本文展示了一种基于球型酶的杂化纳米花(EHNF),AhlX@Ni(PO),可以很容易地合成,其 AHL(3OC8-HSL)降解活性比游离 AhlX 高 10 倍(一种耐热的 AHL 内酯酶)。此外,它在工作浓度下表现出令人惊讶的稳定性,在室温(25°C)下孵育 40 天后保留了约 100%的活性,在 60°C 下孵育 48 小时后保留了约 80%的活性。此外,与 AhlX 相比,它在复杂生态环境中表现出更好的有机溶剂耐受性和长期稳定性。为了降低成本并简化生产工艺,从 AhlX 和 Ni(PO)的粗上清液中组装合成了 CSA@Ni(PO)。AhlX@Ni(PO)和 CSA@Ni(PO)都能有效减轻病原菌感染。
在这项研究中,我们开发了基于 N-酰基高丝氨酸内酯酶的杂化纳米花作为一种新型高效的生物防治试剂,具有显著的控制效果、出色的环境适应性和耐受性。有望克服二十多年来存在的稳定性差和环境耐受能力有限的瓶颈问题,开创 EHNF 在生物防治领域的实际应用。