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通过固定在低分子量壳聚糖纳米粒子上提高海藻酸盐裂解酶的热稳定性和抗生物膜活性。

Enhancing the Thermo-Stability and Anti-Biofilm Activity of Alginate Lyase by Immobilization on Low Molecular Weight Chitosan Nanoparticles.

机构信息

Department of Pharmacology, College of Basic Medicine, Qingdao University, Qingdao 266071, China.

Molecular Cancer Biology Laboratory, Cellular Heterogeneity Research Center, Department of Biosystem, Sookmyung Women's University, Hyochangwon gil-52, Yongsan-Gu, Seoul 140-742, Korea.

出版信息

Int J Mol Sci. 2019 Sep 14;20(18):4565. doi: 10.3390/ijms20184565.

Abstract

Bacterial biofilm causes severe antibiotic resistance. An extracellular polymeric substance (EPS) is the main component in the bacterial biofilm. Alginate is a key EPS component in the biofilm of and responsible for surface adhesion and stabilization of biofilm. Alginate lyase has emerged as an efficient therapeutic strategy targeting to degrade the alginate in the biofilm of . However, the application of this enzyme is limited by its poor stability. In this study, chitosan nanoparticles (CS-NPs) were synthesized using low molecular weight chitosan and alginate lyase Aly08 was immobilized on low molecular weight chitosan nanoparticles (AL-LMW-CS-NPs). As a result, the immobilization significantly enhanced the thermal stability and reusability of Aly08. In addition, compared with free Aly08, the immobilized AL-LMW-CS-NPs exhibited higher efficiency in inhibiting biofilm formation and interrupting the established mature biofilm of , which could reduce its biomass and thickness confirmed by confocal microscopy. Moreover, the biofilm disruption greatly increased the antibiotic sensitivity of . This research will contribute to the further development of alginate lyase as an anti-biofilm agent.

摘要

细菌生物膜导致严重的抗生素耐药性。胞外聚合物质(EPS)是细菌生物膜的主要成分。藻酸盐是 生物膜中关键的 EPS 成分,负责表面附着和生物膜的稳定。藻酸盐裂解酶已成为一种针对降解生物膜中藻酸盐的有效治疗策略。然而,由于其稳定性差,该酶的应用受到限制。在本研究中,使用低分子量壳聚糖合成了壳聚糖纳米颗粒(CS-NPs),并将 Aly08 固定在低分子量壳聚糖纳米颗粒(AL-LMW-CS-NPs)上。结果表明,固定化显著提高了 Aly08 的热稳定性和可重复使用性。此外,与游离的 Aly08 相比,固定化的 AL-LMW-CS-NPs 能够更有效地抑制生物膜的形成,并中断 生物膜的形成和成熟,这可以通过共聚焦显微镜证实减少其生物量和厚度。此外,生物膜的破坏大大增加了 的抗生素敏感性。这项研究将有助于进一步开发藻酸盐裂解酶作为一种抗生物膜剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06c5/6770906/fd65e95683c2/ijms-20-04565-g001.jpg

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