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壳聚糖酶功能化 UiO-66 骨架纳米颗粒对多药耐药性耳念珠菌有效。

Chitinase-functionalized UiO-66 framework nanoparticles active against multidrug-resistant Candida Auris.

机构信息

Department of Chemistry of Natural and Microbial Products, Pharmaceutical and Drug Industries Research Institute, National Research Centre, P.O. 12622, 33 El Bohouth Street, Dokki, Giza, Egypt.

Applied Organic Chemistry Department, Chemical Industries Research Institute, National Research Centre, 33 EL Buhouth St, Dokki, Giza, 12622, Egypt.

出版信息

BMC Microbiol. 2024 Jul 20;24(1):269. doi: 10.1186/s12866-024-03414-1.

Abstract

Candida auris (C. auris) is a yeast that has caused several outbreaks in the last decade. Cell wall chitin plays a primary role in the antifungal resistance of C. auris. Herein, we investigated the potential of chitinase immobilized with UiO-66 to act as a potent antifungal agent against C. auris. Chitinase was produced from Talaromyces varians SSW3 in a yield of 8.97 U/g dry substrate (ds). The yield was statistically enhanced to 120.41 U/g ds by using Plackett-Burman and Box-Behnken design. We synthesized a UiO-66 framework that was characterized by SEM, TEM, XRD, FTIR, a particle size analyzer, and a zeta sizer. The produced framework had a size of 70.42 ± 8.43 nm with a uniform cubic shape and smooth surface. The produced chitinase was immobilized on UiO-66 with an immobilization yield of 65% achieved after a 6 h loading period. The immobilization of UiO-66 increased the enzyme activity and stability, as indicated by the obtained K and T values. Furthermore, the hydrolytic activity of chitinase was enhanced after immobilization on UiO-66, with an increase in the V and a decrease in the K of 2- and 38-fold, respectively. Interestingly, the antifungal activity of the produced chitinase was boosted against C. auris by loading the enzyme on UiO-66, with an MIC of 0.89 ± 0.056 U/mL, compared to 5.582 ± 0.57 U/mL for the free enzyme. This study offers a novel promising alternative approach to combat the new emerging pathogen C. auris.

摘要

耳念珠菌(C. auris)是一种酵母菌,在过去十年中引发了多次爆发。细胞壁几丁质在耳念珠菌的抗真菌耐药性中起主要作用。在此,我们研究了固定化几丁质酶与 UiO-66 结合作为抗耳念珠菌有效抗真菌剂的潜力。几丁质酶是从 Talaromyces varians SSW3 中以 8.97 U/g 干底物(ds)的产率生产的。通过使用 Plackett-Burman 和 Box-Behnken 设计,产率统计上提高到 120.41 U/g ds。我们合成了一种 UiO-66 骨架,其通过 SEM、TEM、XRD、FTIR、粒度分析仪和zeta 粒径仪进行了表征。所生产的骨架具有 70.42 ± 8.43 nm 的尺寸,呈均匀的立方形状且表面光滑。所生产的几丁质酶通过 UiO-66 进行固定化,在 6 小时的加载期后达到 65%的固定化产率。UiO-66 的固定化提高了酶的活性和稳定性,这反映在获得的 K 和 T 值上。此外,几丁质酶在固定化于 UiO-66 后水解活性增强,V 值增加了 2 倍,K 值降低了 38 倍。有趣的是,通过将酶加载到 UiO-66 上,生产的几丁质酶对耳念珠菌的抗真菌活性得到增强,MIC 为 0.89 ± 0.056 U/mL,而游离酶的 MIC 为 5.582 ± 0.57 U/mL。本研究提供了一种对抗新出现的病原体耳念珠菌的有前途的新方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c60/11264975/624e08b29c60/12866_2024_3414_Fig1_HTML.jpg

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