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杠柳种子蛋白的鉴定:通过 LC-ESI-MS/MS 分析及对白色念珠菌和克柔念珠菌的生物技术潜力

Luffa operculata seed proteins: Identification by LC-ESI-MS/MS and biotechnological potential against Candida albicans and C. krusei.

机构信息

Plant Biochemistry and Physiology Laboratory, State University of Rio Grande of North, Mossoró, Rio Grande do Norte, Brazil.

Plant Biochemistry and Physiology Laboratory, State University of Rio Grande of North, Mossoró, Rio Grande do Norte, Brazil; Department of Biochemistry and Molecular Biology, Laboratory of Plant Defense Proteins, Federal University of Ceará, Fortaleza, Ceará, Brazil.

出版信息

Anal Biochem. 2022 Oct 15;655:114851. doi: 10.1016/j.ab.2022.114851. Epub 2022 Aug 14.

Abstract

L

operculata is a plant commonly found in the North and Northeast of Brazil. Although the regional population knows its medicinal potential, there are few scientific studies about its antimicrobial potential. Thus, this study aimed to characterize the proteins from L. operculata seeds extracted using different solutions and evaluate their antimicrobial potentials. The protein extracts obtained with NaCl and sodium acetate buffer presented the best inhibitory activities against Candida albicans and C. krusei. The study of the mechanism of action revealed proteins from L. operculata seeds induced pore formation on the membrane and ROS overaccumulation. Scanning Electron Microscopy images also showed severe morphological changes in Candida albicans and C. krusei. Proteins from L.operculata seeds did not show antibacterial activity. The enzymatic assays revealed the presence of proteolytic enzymes, serine and cysteine protease inhibitors, and chitinases in both protein extracts. Proteomic analysis by LC-ESI-MS/MS identified 57 proteins related to many biological processes, such as defense to (a)biotic stress, energetic metabolism, protein folding, and nucleotide metabolism. In conclusion, the L. operculata seed proteins have biotechnological potential against the human pathogenic yeasts Candida albicans and C. krusei.

摘要

巴西北部和东北部常见的植物有桃叶珊瑚。尽管该地区的人们知道它的药用潜力,但关于它的抗菌潜力的科学研究却很少。因此,本研究旨在对使用不同溶液提取的桃叶珊瑚种子中的蛋白质进行表征,并评估其抗菌潜力。用 NaCl 和醋酸钠缓冲液提取的蛋白质提取物对白色念珠菌和克柔念珠菌表现出最好的抑制活性。作用机制的研究表明,桃叶珊瑚种子中的蛋白质在细胞膜上诱导孔形成和 ROS 过度积累。扫描电子显微镜图像还显示白色念珠菌和克柔念珠菌的严重形态变化。桃叶珊瑚种子的蛋白质没有表现出抗菌活性。酶活性分析显示两种蛋白质提取物中均存在蛋白酶、丝氨酸和半胱氨酸蛋白酶抑制剂以及几丁质酶。LC-ESI-MS/MS 的蛋白质组学分析鉴定了 57 种与许多生物过程相关的蛋白质,如防御(非生物)胁迫、能量代谢、蛋白质折叠和核苷酸代谢。总之,桃叶珊瑚种子蛋白具有针对人类致病性酵母白色念珠菌和克柔念珠菌的生物技术潜力。

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