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抗菌肽 MaltAtt-1 在 GS115 中用于防治松材线虫的优化和稳定性评估。

Optimization and Stability Assessment of Antimicrobial Peptide MaltAtt-1 in GS115 for the Control of Pine Wood Nematode.

机构信息

Forestry College, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

出版信息

Int J Mol Sci. 2024 Aug 6;25(16):8555. doi: 10.3390/ijms25168555.

Abstract

MaltAtt-1 is an antimicrobial peptide isolated from with nematocidal activity against pine wood nematode. In this study, a eukaryotic expression system based on GS115 was established, and its secretory expression of MaltAtt-1 was realized. The basic properties and secondary and tertiary structures of the antimicrobial peptide MaltAtt-1 were identified by bioinformatics analysis. MaltAtt-1 is a hydrophilic stable protein, mainly composed of an α-helix (Hh), β-folds (Ee), and irregular curls (Cc). The optimal fermentation conditions for MaltAtt-1 were determined by a single-factor test and the Box-Behnken response surface method, including an induction time of 72 h, induction temperature of 30 °C, culture medium of pH 7.6, methanol volume fraction of 2.0%, and an initial glycerol concentration of 1%. The stability of MaltAtt-1 indicated its resistant to UV irradiation and repeated freezing and thawing, but the antibacterial activity decreased significantly under the influence of high temperature and a strong acid and base, and it decreased significantly to 1.1 cm and 0.83 cm at pH 2.0 and pH 10.0, respectively. The corrected mortality of . achieved 71.94% in 3 h at a concentration of 300 mg·L MaltAtt-1 exposure. The results provide a theoretical basis for the antimicrobial peptide MaltAtt-1 to become a new green and efficient nematicide.

摘要

MaltAtt-1 是一种从 中分离出的具有杀线虫活性的抗菌肽。本研究建立了基于 GS115 的真核表达系统,实现了 MaltAtt-1 的分泌表达。通过生物信息学分析鉴定了抗菌肽 MaltAtt-1 的基本性质和二级、三级结构。MaltAtt-1 是一种亲水稳定的蛋白质,主要由 α-螺旋(Hh)、β-折叠(Ee)和不规则卷曲(Cc)组成。通过单因素试验和 Box-Behnken 响应面法确定了 MaltAtt-1 的最佳发酵条件,包括诱导时间 72 h、诱导温度 30°C、培养基 pH7.6、甲醇体积分数 2.0%和初始甘油浓度 1%。MaltAtt-1 的稳定性表明其能抵抗紫外线辐射和反复冻融,但在高温和强酸强碱的影响下,抗菌活性显著下降,在 pH2.0 和 pH10.0 时,抗菌活性分别下降到 1.1cm 和 0.83cm。浓度为 300mg·L 的 MaltAtt-1 暴露 3 小时后,对. 的校正死亡率达到 71.94%。结果为抗菌肽 MaltAtt-1 成为一种新型绿色高效杀线虫剂提供了理论基础。

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