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利用噬菌体展示技术鉴定新型对羟基苯甲酸酯结合肽。

Identification of novel paraben-binding peptides using phage display.

机构信息

Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju-si, Jeollabuk-do, 54896, Republic of Korea.

Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.

出版信息

Environ Pollut. 2020 Dec;267:115479. doi: 10.1016/j.envpol.2020.115479. Epub 2020 Aug 28.

Abstract

Parabens are alkyl esters of 4-hydroxybenzoic acid, which is derived from a family of synthetic esters of p-hydroxybenzoic acid. Among all the kinds of paraben, two parabens (methyl paraben, MP; and n-propyl paraben, PP) are the most generally used as preservatives in personal care products, such as cosmetics, pharmaceuticals, and food also, and are often presented together. However, a number of studies have reported that the toxicity of parabens affects the water environment, and human as well. This study utilized M13 phage display technology to provide easy, efficient, and relatively inexpensive methods to identify peptides that bind to MP and PP, respectively, to remove in wastewater. At first, biopanning was performed, to sort MP and PP specific binding phages, and three cases of experiment, including negative control (NC), which could sort unspecific binding phage, were conducted at the same time. Phage binding affinity tests were substituted by concentration reduction using antibody conjugated magnetic beads, and paraben concentration was measured by HPLC. Analysis showed that the MP concentration reduction of 38% was the highest in M4 phage, while the PP concentration reduction of 44% was the highest in P3 phage. We successfully screened two peptides specific to MP and PP, namely, MP4 and PP3, respectively; the results showed that the MP concentration reduction in MP4 was the highest at 44%, and the PP concentration reduction in PP3 was the highest at 39%, and their specificity was measured by the capture rate between target and control. In conclusion, the phage display technique shows applicability to the removal of parabens in water; furthermore, it also shows the possibility of the detection or removal of other chemicals.

摘要

对羟基苯甲酸酯是对羟基苯甲酸的烷基酯,对羟基苯甲酸则是对羟基苯甲酸酯家族的一种人工合成酯。在所有对羟基苯甲酸酯中,两种对羟基苯甲酸酯(对羟基苯甲酸甲酯、MP 和对羟基苯甲酸丙酯、PP)最常被用作个人护理产品(如化妆品、药品和食品)中的防腐剂,并且经常同时出现。然而,许多研究报告称,对羟基苯甲酸酯的毒性会影响水环境和人类。本研究利用 M13 噬菌体展示技术,提供了简便、高效和相对廉价的方法,以识别分别与 MP 和 PP 结合的肽,从而去除废水中的这些防腐剂。首先进行了生物淘选,以筛选出与 MP 和 PP 特异性结合的噬菌体,同时进行了三个实验,包括阴性对照(NC),以筛选出非特异性结合的噬菌体。噬菌体结合亲和力测试被抗体偶联磁珠的浓度降低所取代,并且通过 HPLC 测量了对羟基苯甲酸酯的浓度。分析表明,M4 噬菌体对 MP 的浓度降低率最高,达到 38%,而 P3 噬菌体对 PP 的浓度降低率最高,达到 44%。我们成功筛选出两种分别与 MP 和 PP 特异性结合的肽,即 MP4 和 PP3,结果表明,MP4 对 MP 的浓度降低率最高,达到 44%,而 PP3 对 PP 的浓度降低率最高,达到 39%,并且通过目标与对照之间的捕获率来测量其特异性。总之,噬菌体展示技术显示出在水中去除对羟基苯甲酸酯的适用性;此外,它还显示了检测或去除其他化学物质的可能性。

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