Taveira Gabriel B, Mathias Luciana S, da Motta Olney V, Machado Olga L T, Rodrigues Rosana, Carvalho André O, Teixeira-Ferreira André, Perales Jonas, Vasconcelos Ilka M, Gomes Valdirene M
Center equivalent to department, Universidade Estadual do Norte Fluminense, Centro de Biociências e Biotecnologia, Campos dos Goytacazes, RJ, Brazil.
Biopolymers. 2014 Jan;102(1):30-9. doi: 10.1002/bip.22351.
Plants defend themselves against pathogens with production of antimicrobial peptides (AMPs). Herein we describe the discovery of a new antifungal and antibacterial peptide from fruits of Capsicum annuum that showed similarity to an already well characterized family of plant AMPs, thionins. Other fraction composed of two peptides, in which the major peptide also showed similarity to thionins. Among the obtained fractions, fraction 1, which is composed of a single peptide of 7 kDa, was sequenced by Edman method and its comparative sequence analysis in database (nr) showed similarity to thionin-like peptides. Tests against microorganisms, fraction 1 presented inhibitory activity to the cells of yeast Saccharomyces cerevisiae, Candida albicans, and Candida tropicalis and caused growth reduction to the bacteria species Escherichia coli and Pseudomonas aeruginosa. Fraction 3 caused inhibitory activity only for C. albicans and C. tropicalis. This fraction was composed of two peptides of ∼7 and 10 kDa, and the main protein band correspondent to the 7 kDa peptide, also showed similarity to thionins. This plasma membrane permeabilization assay demonstrates that the peptides present in the fractions 1 and 3 induced changes in the membranes of all yeast strains, leading to their permeabilization. Fraction 1 was capable of inhibiting acidification of the medium of glucose-induced S. cerevisiae cells 78% after an incubation time of 30 min, and opposite result was obtained for C. albicans. Experiments demonstrate that the fraction 1 and 3 were toxic and induced changes in the membranes of all yeast strains, leading to their permeabilization.
植物通过产生抗菌肽(AMPs)来抵御病原体。在此,我们描述了从辣椒果实中发现的一种新的抗真菌和抗菌肽,它与一个已被充分表征的植物AMPs家族——硫堇蛋白相似。其他部分由两种肽组成,其中主要的肽也与硫堇蛋白相似。在所获得的部分中,由一个7 kDa的单一肽组成的部分1通过埃德曼方法进行了测序,其在数据库(nr)中的比较序列分析显示与类硫堇蛋白肽相似。针对微生物的测试表明,部分1对酿酒酵母、白色念珠菌和热带念珠菌的细胞具有抑制活性,并导致大肠杆菌和铜绿假单胞菌这两种细菌的生长减少。部分3仅对白色念珠菌和热带念珠菌具有抑制活性。该部分由约7 kDa和10 kDa的两种肽组成,与7 kDa肽相对应的主要蛋白条带也与硫堇蛋白相似。这种质膜通透性测定表明,部分1和3中存在的肽诱导了所有酵母菌株膜的变化,导致其通透性增加。部分1在孵育30分钟后能够抑制葡萄糖诱导的酿酒酵母细胞培养基酸化78%,而白色念珠菌则得到相反的结果。实验表明,部分1和3具有毒性,并诱导了所有酵母菌株膜的变化,导致其通透性增加。