Rahman Mahmudur, Browne Jessica J, Van Crugten Jacoba, Hasan Md Fahim, Liu Lei, Barkla Bronwyn J
Southern Cross Plant Science, Southern Cross University, Lismore, NSW, Australia.
School of Health and Human Sciences, Southern Cross University, Bilinga, QLD, Australia.
Front Pharmacol. 2020 Sep 8;11:1340. doi: 10.3389/fphar.2020.01340. eCollection 2020.
In addition to their use as an edible oil and condiment crop, mustard and rapeseed ( L., (L.) Czern., (L.) W.D.J.Koch, L. and L.) have been commonly used in traditional medicine for relieving pain, coughs and treating infections. The seeds contain high amounts of oil, while the remaining by-product meal after oil extraction, about 40% of seed dry weight, has a low value despite its high protein-content (~85%). The seed storage proteins (SSP) 2S albumin-type napin and 12S globulin-type cruciferin are the two predominant proteins in the seeds and show potential for value adding to the waste stream; however, information on their biological activities is scarce. In this study, purified napin and cruciferin were tested using , molecular docking, and approaches for their bioactivity as antimicrobial peptides.
The 3D-structure of 2S albumin and 12S globulin storage proteins from were investigated to predict antimicrobial activity employing an antimicrobial peptide database survey. To gain deeper insights into the potential antimicrobial activity of these SSP, molecular docking was performed. The purified cruciferin and napin were then tested against both Gram-positive and Gram-negative bacteria for antimicrobial activity by disc diffusion and microdilution antimicrobial susceptibility testing.
analysis demonstrated both SSP share similar 3D-structure with other well studied antimicrobial proteins. Molecular docking revealed that the proteins exhibited high binding energy to bacterial enzymes. Cruciferin and napin proteins appeared as a double triplet and a single doublet, respectively, following SDS-PAGE. SDS-PAGE and Western blotting also confirmed the purity of the protein samples used for assessment of antimicrobial activity. Antimicrobial susceptibility testing provided strong evidence for antimicrobial activity for the purified napin protein; however, cruciferin showed no antimicrobial activity, even at the highest dose applied.
and molecular docking results presented evidence for the potential antimicrobial activity of rapeseed cruciferin and napin SSP. However, only the antimicrobial activity of napin was confirmed. These findings warrant further investigation of this SSP protein as a potential new agent against infectious disease.
除了用作食用油和调味品作物外，芥菜和油菜籽（甘蓝型油菜、白菜型油菜、芥菜型油菜、埃塞俄比亚芥和野芥）在传统医学中还常用于缓解疼痛、止咳和治疗感染。种子含有大量油脂，而榨油后剩余的副产品粕约占种子干重的40%，尽管其蛋白质含量高（约85%），但价值较低。种子贮藏蛋白（SSP）2S白蛋白型napin和12S球蛋白型cruciferin是种子中的两种主要蛋白质，具有为废物流增值的潜力；然而，关于它们生物活性的信息却很少。在本研究中，使用抗菌肽数据库调查、分子对接和抗菌药敏试验方法对纯化的napin和cruciferin的抗菌肽生物活性进行了测试。
通过抗菌肽数据库调查研究甘蓝型油菜2S白蛋白和12S球蛋白贮藏蛋白的三维结构，以预测其抗菌活性。为了更深入了解这些SSP的潜在抗菌活性，进行了分子对接。然后通过纸片扩散法和微量稀释抗菌药敏试验，对纯化的cruciferin和napin针对革兰氏阳性菌和革兰氏阴性菌的抗菌活性进行测试。
分析表明，两种SSP与其他经过充分研究的抗菌蛋白具有相似的三维结构。分子对接显示，这些蛋白质与细菌酶具有高结合能。SDS-PAGE电泳后，cruciferin蛋白呈现双三联体，napin蛋白呈现单双峰。SDS-PAGE和蛋白质印迹法也证实了用于评估抗菌活性的蛋白质样品的纯度。抗菌药敏试验为纯化的napin蛋白的抗菌活性提供了有力证据；然而，即使在最高剂量下，cruciferin也没有显示出抗菌活性。
抗菌肽数据库调查和分子对接结果为油菜籽cruciferin和napin SSP的潜在抗菌活性提供了证据。然而，仅证实了napin的抗菌活性。这些发现值得进一步研究这种SSP蛋白作为一种潜在的抗传染病新药物。
