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