Taniguchi Masayuki, Saito Kengo, Nomoto Takafumi, Namae Toshiki, Ochiai Akihito, Saitoh Eiichi, Tanaka Takaaki
Department of Materials Science and Technology, Graduate School of Science and Technology, Niigata University, Niigata, 950-2181, Japan.
Center for Transdisciplinary Research, Niigata University, Niigata, 950-2181, Japan.
Biopolymers. 2017 Jul;108(4). doi: 10.1002/bip.23023.
In this study, we identified and chemically synthesized three cationic and amphipathic peptides (Glycinin-17, BCAS-16, and BCBS-11) from soybean proteins. These peptides had high isoelectric points, high positive net charges, and included multiple hydrophobic amino acids. Subsequently, we identified multiple functions of these peptides, including antimicrobial, lipopolysaccharide-neutralizing, and angiogenic activities, and examined their cytotoxic activities against mammalian red blood cells. Glycinin-17, BCAS-16, and BCBS-11 exhibited antimicrobial activity against Porphyromonas gingivalis and Candida albicans whereas Glycinin-17 did not possess antimicrobial effects on Propionibacterium acnes and Streptococcus mutans. Membrane-depolarization assays and flow cytometric analyses showed that the antimicrobial properties of Glycinin-17, BCAS-16, and BCBS-11 against P. gingivalis, P. acnes, and S. mutans were dependent on membrane-disrupting potential. In contrast, major antimicrobial activities of these peptides against C. albicans were dependent on interactions with targets other than cell membranes. Furthermore, chromogenic Limulus amebocyte lysate assays showed that 50% effective concentrations (EC , 0.12-0.31 μM) of these three peptides neutralize LPS with similar potency (EC : 0.11 μM) to that of polymyxin B. Moreover, tube-formation assays in human umbilical vein endothelial cells showed similar angiogenic activities of the three peptides as that following treatment with LL-37. Although BCAS-16 exhibited hemolytic activity, the rate of hemolysis for Glycinin-17 and BCBS-11 in the presence of 500-μM Glycinin-17 and BCBS-11 was less than 2%. These results demonstrate that cationic and amphipathic peptides from soybean proteins, particularly Glycinin-17 and BCBS-11, have potential as multifunctional ingredients for healthcare applications.
在本研究中，我们从大豆蛋白中鉴定并化学合成了三种阳离子两亲性肽（大豆球蛋白-17、BCAS-16和BCBS-11）。这些肽具有高的等电点、高的正净电荷，并且包含多个疏水氨基酸。随后，我们鉴定了这些肽的多种功能，包括抗菌、中和脂多糖和血管生成活性，并检测了它们对哺乳动物红细胞的细胞毒性活性。大豆球蛋白-17、BCAS-16和BCBS-11对牙龈卟啉单胞菌和白色念珠菌具有抗菌活性，而大豆球蛋白-17对痤疮丙酸杆菌和变形链球菌没有抗菌作用。膜去极化测定和流式细胞术分析表明，大豆球蛋白-17、BCAS-16和BCBS-11对牙龈卟啉单胞菌、痤疮丙酸杆菌和变形链球菌的抗菌特性取决于膜破坏潜力。相反，这些肽对白色念珠菌的主要抗菌活性取决于与细胞膜以外的靶点的相互作用。此外，显色鲎试剂法检测表明，这三种肽的50%有效浓度（EC50，0.12 - 0.31 μM）中和脂多糖的效力与多粘菌素B相似（EC50：0.11 μM）。此外，人脐静脉内皮细胞中的管形成试验表明，这三种肽的血管生成活性与用LL-37处理后的活性相似。尽管BCAS-16表现出溶血活性，但在存在500 μM大豆球蛋白-17和BCBS-11的情况下，大豆球蛋白-17和BCBS-11的溶血率低于2%。这些结果表明，来自大豆蛋白的阳离子两亲性肽，特别是大豆球蛋白-17和BCBS-11，具有作为医疗保健应用多功能成分的潜力。
