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利用疏水蛋白嵌合蛋白开发抗菌表面。

Development of anti-bacterial surfaces using a hydrophobin chimeric protein.

机构信息

Department of Chemical Sciences, University of Naples Federico II, Napoli, Italy.

Institute of Protein Biochemistry, CNR, Napoli, Italy.

出版信息

Int J Biol Macromol. 2020 Dec 1;164:2293-2300. doi: 10.1016/j.ijbiomac.2020.07.301. Epub 2020 Aug 5.

Abstract

The search for new approaches for developing antimicrobial surfaces is a challenge of great urgency to prevent and control microbial growth on surfaces. The strategy herein proposed relies on the design of a new, simple and general tool for creating antimicrobial surfaces, based on a hydrophobin chimeric protein which fuses the adhesive self-assembling class I hydrophobin Vmh2 from Pleurotus ostreatus to the human antimicrobial peptide LL-37. The recombinant LL37-Vmh2 protein displayed both the adhesive and the antimicrobic properties of its members, and when deposited on polystyrene surface, a positive effect due to the fusion was observed in terms of both efficacy and versatility of the coating. Indeed, the chimeric protein significantly enlarges the range of pathogens affected by Vmh2 layer rendering it able to inhibit three Gram-positive and two Gram-negative pathogens, selected among the renowned biofilm producer bacteria. Confocal Laser Scanning Microscopy analysis performed on Staphylococcus epidermidis biofilms formed on coated surfaces proved that, besides inhibiting biofilm formation, the LL37-Vmh2 coating also displayed biocidal activity, since dead cells were present in the biofilm layer. The reported results open new perspectives in various fields of application of LL37, and of antimicrobial peptides in general. LL37-Vmh2 increases the inventory of chimeric hydrophobins, further proving their effectiveness and versatility in surface functionalization.

摘要

寻找开发抗菌表面的新方法是预防和控制表面微生物生长的一项非常紧迫的挑战。本文提出的策略依赖于设计一种新的、简单的和通用的工具来创建抗菌表面,该工具基于融合了来自糙皮侧耳(Pleurotus ostreatus)的具有粘附性的自组装 I 类疏水蛋白 Vmh2 与人抗菌肽 LL-37 的嵌合疏水蛋白。重组的 LL37-Vmh2 蛋白表现出其成员的粘附和抗菌特性,并且当沉积在聚苯乙烯表面上时,由于融合,观察到涂层的功效和通用性方面都有积极的效果。事实上,嵌合蛋白显著扩大了 Vmh2 层影响的病原体范围,使其能够抑制三种革兰氏阳性菌和两种革兰氏阴性菌,这些病原体是从著名的生物膜产生菌中选择的。在涂覆表面上形成的表皮葡萄球菌生物膜上进行的共焦激光扫描显微镜分析证明,除了抑制生物膜形成外,LL37-Vmh2 涂层还显示出杀菌活性,因为生物膜层中存在死亡细胞。所报道的结果为 LL37 以及一般抗菌肽在各个应用领域开辟了新的前景。LL37-Vmh2 增加了嵌合疏水蛋白的种类,进一步证明了它们在表面功能化方面的有效性和通用性。

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