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海洋源铜绿假单胞菌(DKB1)鼠李糖脂的光谱学和生物医学特性研究。

Investigation on spectral and biomedical characterization of rhamnolipid from a marine associated bacterium Pseudomonas aeruginosa (DKB1).

机构信息

Department of Microbiology, K.R. College of Arts and Science, Kovilpatti, Tamilnadu, 628503, India.

MNP Laboratory, Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam, Kanyakumari, Tamilnadu, 629502, India.

出版信息

Arch Microbiol. 2021 Jul;203(5):2297-2314. doi: 10.1007/s00203-021-02220-x. Epub 2021 Mar 1.

Abstract

Bio-surfactants are a principal group of significant molecules obtained from the microbial sources expressed with distinctive characteristics like biodegradation of hydrocarbons and also have different biomedical properties. The present investigation aims to assess the biomedical properties of synthesized bio-surfactant, rhamnolipid from Pseudomonas aeruginosa (DKB1) under in vitro conditions. The candidate bacterium P. aeruginosa (DKB1) was isolated from oil-polluted fishing harbors of Kanyakumari coast. Initially, the bio-surfactant production by this candidate strain was confirmed by oil displacement assay, hemolytic assay, drop collapse assay and emulsification index. Further, the production of bio-surfactant was achieved through submerged fermentation process using Bushnell-Haas mineral salts medium supplemented with 2% olive oil. The yield of the bio-surfactant was attained as 2.4 g/l and confirmed as rhamnolipid through blue agar plate assay; further, the extracted rhamnolipid was purified and characterized through standard procedures. In stability studies, the rhamnolipid could withstand up to pH 12, temperature 100 °C and 15% of NaCl concentration. The biomedical application of rhamnolipid (30 μg ml) was determined by antibacterial, antioxidant and cytotoxic studies. It exhibited a maximum growth inhibition against Bacillus subtilis (26 mm) with the MIC value of 8 μg ml. In antioxidant test, rhamnolipid expressed significant (P < 0.0001) inhibition of total reducing power (44.11%), DPPH activity (61.60%), hydroxyl radical (83.30%) and nitric oxide (51.86%) scavenging ability at 100 μg mlwith the respective IC50 values of 130.50, 77.18, 52.08 and 95.43 μg ml. The anticancer activity of the rhamnolipid was assessed with the help of MTT test against MCF-7, HT-29 and E-143 cancer cell lines individually, and the viability of the cells was observed, respectively, as 10.24, 17.66 and 13.50% at 250 μg mlconcentration with the respective IC50 values of 140.2, 81.02 and 138.9 μg ml. From the results, it could be concluded that the rhamnolipid produced by P. aeruginosa (DKB1) isolated from oil-polluted area has effective biomedical properties.

摘要

生物表面活性剂是一类主要的分子,来源于微生物,具有独特的特性,如烃类的生物降解,并且具有不同的生物医学特性。本研究旨在评估从铜绿假单胞菌(DKB1)合成的生物表面活性剂鼠李糖脂的体外生物医学特性。候选菌铜绿假单胞菌(DKB1)是从楠奇库马里海岸的石油污染渔港中分离出来的。最初,通过油置换试验、溶血试验、滴崩溃试验和乳化指数证实了该候选菌株的生物表面活性剂的产生。进一步,通过在 Bushnell-Haas 无机盐培养基中添加 2%橄榄油的液体发酵过程来实现生物表面活性剂的产生。生物表面活性剂的产率达到 2.4g/L,并通过蓝色琼脂平板试验确认为鼠李糖脂;进一步,通过标准程序对提取的鼠李糖脂进行纯化和表征。在稳定性研究中,鼠李糖脂可耐受 pH 值 12、温度 100°C 和 15%的 NaCl 浓度。通过抗菌、抗氧化和细胞毒性研究来确定鼠李糖脂的生物医学应用。它对枯草芽孢杆菌(26mm)的最大生长抑制作用,其 MIC 值为 8μg/ml。在抗氧化试验中,鼠李糖脂表达了显著的(P<0.0001)总还原力(44.11%)、DPPH 活性(61.60%)、羟基自由基(83.30%)和一氧化氮(51.86%)清除能力,在 100μg/ml 时,相应的 IC50 值分别为 130.50、77.18、52.08 和 95.43μg/ml。通过 MTT 试验分别评估了鼠李糖脂对 MCF-7、HT-29 和 E-143 癌细胞系的抗癌活性,并观察到细胞的活力分别为 250μg/ml 浓度时的 10.24%、17.66%和 13.50%,相应的 IC50 值分别为 140.2、81.02 和 138.9μg/ml。从结果可以得出结论,从石油污染地区分离的铜绿假单胞菌(DKB1)产生的鼠李糖脂具有有效的生物医学特性。

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