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几丁聚糖载固定化木瓜蛋白酶对口腔致病菌生物膜的影响。

The Effect of Ficin Immobilized on Carboxymethyl Chitosan on Biofilms of Oral Pathogens.

机构信息

Institute of Fundamental Medicine and Biology, Kazan (Volga Region) Federal University, 420008 Kazan, Russia.

Department of Biophysics and Biotechnology, Voronezh State University, 394018 Voronezh, Russia.

出版信息

Int J Mol Sci. 2023 Nov 8;24(22):16090. doi: 10.3390/ijms242216090.

Abstract

In the last decade, Ficin, a proteolytic enzyme extracted from the latex sap of the wild fig tree, has been widely investigated as a promising tool for the treatment of microbial biofilms, wound healing, and oral care. Here we report the antibiofilm properties of the enzyme immobilized on soluble carboxymethyl chitosan (CMCh) and CMCh itself. Ficin was immobilized on CMCh with molecular weights of either 200, 350 or 600 kDa. Among them, the carrier with a molecular weight of 200 kDa bound the maximum amount of enzyme, binding up to 49% of the total protein compared to 19-32% of the total protein bound to other CMChs. Treatment with pure CMCh led to the destruction of biofilms formed by , , , and , while no apparent effect on was observed. A soluble Ficin was less efficient in the destruction of the biofilms formed by and . By contrast, treatment with CMCh200-immobilized Ficin led to a significant reduction of the biofilms of the primary colonizers and In model biofilms obtained by the inoculation of swabs from teeth of healthy volunteers, the destruction of the biofilm by both soluble and immobilized Ficin was observed, although the degree of the destruction varied between artificial plaque samples. Nevertheless, combined treatment of oral biofilm by enzyme and chlorhexidine for 3 h led to a significant decrease in the viability of biofilm-embedded cells, compared to solely chlorhexidine application. This suggests that the use of either soluble or immobilized Ficin would allow decreasing the amount and/or concentration of the antiseptics required for oral care or improving the efficiency of oral cavity sanitization.

摘要

在过去的十年中,无花果凝乳酶作为一种从野生无花果树乳胶中提取的蛋白水解酶,因其在治疗微生物生物膜、伤口愈合和口腔护理方面的广阔应用前景而受到广泛研究。本研究报道了固定在可溶性羧甲基壳聚糖(CMCh)上的酶和 CMCh 本身的抗生物膜特性。无花果凝乳酶通过与分子量为 200、350 或 600 kDa 的 CMCh 结合而被固定。其中,分子量为 200 kDa 的载体结合了最多的酶,与其他 CMCh 相比,结合了高达 49%的总蛋白,而结合了 19-32%的总蛋白。纯 CMCh 处理可破坏 、 、 、 和 形成的生物膜,而对 没有明显影响。纯可溶性无花果凝乳酶对 、 形成的生物膜的破坏效果较差。相比之下,CMCh200-固定化无花果凝乳酶处理可显著减少 和 的生物膜。在通过接种来自健康志愿者牙齿拭子获得的模型生物膜中,观察到可溶性和固定化无花果凝乳酶均可破坏生物膜,尽管在人工菌斑样本之间破坏程度有所不同。然而,酶和洗必泰联合处理 3 小时可显著降低生物膜内细胞的活力,与单独使用洗必泰相比。这表明,无论是使用可溶性还是固定化的无花果凝乳酶,都可以减少口腔护理所需的防腐剂的用量和/或浓度,或提高口腔消毒的效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dcea/10671066/6efc1550c93f/ijms-24-16090-g001.jpg

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