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互补光谱研究与副地衣芽孢杆菌 ND2 产生的蔗果型果聚糖的潜在活性

Complementary spectroscopy studies and potential activities of levan-type fructan produced by Bacillus paralicheniformis ND2.

机构信息

Department of Botany and Microbiology, Faculty of Science, Kafrelsheikh University, Kafr El Sheikh 33516, Egypt.

National Institute of Oceanography and Fisheries (NIOF), Egypt.

出版信息

Carbohydr Polym. 2023 Jul 1;311:120743. doi: 10.1016/j.carbpol.2023.120743. Epub 2023 Feb 26.

Abstract

This study aimed at the production of marine bacterial exopolysaccharides (EPS) as biodegradable and nontoxic biopolymers, competing the synthetic derivatives, with detailed structural and conformational analyses using spectroscopy techniques. Twelve marine bacterial bacilli were isolated from the seawater of Mediterranean Sea, Egypt, then screened for EPS production. The most potent isolate was identified genetically as Bacillus paralicheniformis ND2 by16S rRNA gene sequence of ~99 % similarity. Plackett-Burman (PB) design identified the optimization conditions of EPS production, which yielded the maximum EPS (14.57 g L) with 1.26-fold increase when compared to the basal conditions. Two purified EPSs namely NRF1 and NRF2 with average molecular weights (Mw¯) of 15.98 and 9.70 kDa, respectively, were obtained and subjected for subsequent analyses. FTIR and UV-Vis reflected their purity and high carbohydrate contents while EDX emphasized their neutral type. NMR identified the EPSs as levan-type fructan composed of β-(2-6)-glycosidic linkage as a main backbone, and HPLC explained that the EPSs composed of fructose. Circular dichroism (CD) suggested that NRF1 and NRF2 had identical structuration with a little variation from the EPS-NR. The EPS-NR showed antibacterial activity with the maximum inhibition against S. aureus ATCC 25923. Furthermore, all the EPSs revealed a proinflammatory action through dose-dependent increment of expression of proinflammatory cytokine mRNAs, IL-6, IL-1β and TNFα.

摘要

本研究旨在生产海洋细菌胞外多糖(EPS)作为可生物降解和无毒的生物聚合物,与合成衍生物竞争,使用光谱技术进行详细的结构和构象分析。从埃及地中海海水分离出 12 株海洋细菌杆菌,然后筛选 EPS 生产。最有效的分离株通过 16S rRNA 基因序列鉴定为~99%相似的 Paralicheniformis ND2。Plackett-Burman(PB)设计确定了 EPS 生产的优化条件,与基础条件相比,EPS(14.57g/L)产量最高增加了 1.26 倍。获得了两种纯化的 EPS,即 NRF1 和 NRF2,其平均分子量(Mw¯)分别为 15.98 和 9.70kDa,并进行了后续分析。FTIR 和 UV-Vis 反映了它们的纯度和高碳水化合物含量,而 EDX 强调了它们的中性类型。NMR 将 EPS 鉴定为以β-(2-6)糖苷键为主链的纤维二糖型果聚糖,HPLC 解释了 EPS 由果糖组成。圆二色性(CD)表明 NRF1 和 NRF2 具有相同的结构,与 EPS-NR 略有差异。EPS-NR 表现出抗菌活性,对 ATCC 25923 的金黄色葡萄球菌的抑制作用最大。此外,所有 EPS 都通过表达促炎细胞因子 mRNAs、IL-6、IL-1β 和 TNFα 的剂量依赖性增加显示出促炎作用。

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