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溶菌酶与表没食子儿茶素没食子酸酯的共轭作用以改善抗菌和抗氧化性能

Conjugation of Lysozyme and Epigallocatechin Gallate for Improving Antibacterial and Antioxidant Properties.

作者信息

Moreno-Vásquez María J, Carretas-Valdez Manuel I, Luque-Alcaraz Ana G, Quintero-Reyes Idania E, Tapia-Hernández José A, Arvizu-Flores Aldo A, Moreno-Córdova Elena N, Graciano-Verdugo Abril Z

机构信息

Departamento de Ciencias Químico Biológicas, Universidad de Sonora, Hermosillo, 83000, México.

Departamento de Ciencias de la Salud, Universidad de Sonora, Cd. Obregón, 85040, México.

出版信息

Curr Microbiol. 2024 Jul 13;81(9):264. doi: 10.1007/s00284-024-03776-9.

Abstract

One of the main interests in the food industry is the preservation of food from spoilage by microorganisms or lipid oxidation. A novel alternative is the development of additives of natural origin with dual activity. In the present study, a chemically modified lysozyme (Lys) with epigallocatechin gallate (EGCG) was developed to obtain a conjugate (Lys-EGCG) with antibacterial/antioxidant activity to improve its properties and increase its application potential. The modification reaction was carried out using a free radical grafting method for the Lys modification reaction, using ascorbic acid and hydrogen peroxide as radical initiators in an aqueous medium. The synthesis of Lys-EGCG conjugate was confirmed by spectroscopic (FT-IR, H-RMN, and XPS) and calorimetry differential scanning (DSC) analyses. The EGCG binding to the Lys biomolecule was quantified by the Folin-Ciocalteu method; the antibacterial activity was evaluated by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MCB) against Staphylococcus aureus and Pseudomonas fluorescens; the antioxidant activity was evaluated by ABTS, DPPH, and FRAP. The spectroscopic results showed that the Lys-EGCG conjugate was successfully obtained, and the DSC analysis revealed a 20 °C increase (P < 0.05) in the denaturation temperature of Lys due to EGCG modification. The EGCG concentration in Lys-EGCG was 97.97 ± 4.7 µmol of EGCG/g of sample. The antibacterial and antioxidant activity of the Lys-EGCG conjugate was higher (P < 0.05) than pure EGCG and Lys. The chemical modification of Lys with EGCG allows for the bioconjugate with a dual function (antibacterial/antioxidant), broadening the range of Lys and EGCG applications to different areas such as food, cosmetic, and pharmaceutical industries.

摘要

食品工业的主要关注点之一是防止食品因微生物或脂质氧化而变质。一种新的替代方法是开发具有双重活性的天然来源添加剂。在本研究中,开发了一种用表没食子儿茶素没食子酸酯(EGCG)化学修饰的溶菌酶(Lys),以获得具有抗菌/抗氧化活性的共轭物(Lys-EGCG),从而改善其性能并增加其应用潜力。修饰反应采用自由基接枝法进行Lys修饰反应,在水介质中使用抗坏血酸和过氧化氢作为自由基引发剂。通过光谱分析(傅里叶变换红外光谱、氢核磁共振光谱和X射线光电子能谱)和差示扫描量热法(DSC)分析证实了Lys-EGCG共轭物的合成。通过福林-酚试剂法对EGCG与Lys生物分子的结合进行了定量;通过对金黄色葡萄球菌和荧光假单胞菌的最低抑菌浓度(MIC)和最低杀菌浓度(MCB)评估抗菌活性;通过ABTS、DPPH和FRAP评估抗氧化活性。光谱结果表明成功获得了Lys-EGCG共轭物,DSC分析显示由于EGCG修饰,Lys的变性温度升高了20℃(P<0.05)。Lys-EGCG中EGCG的浓度为97.97±4.7μmol EGCG/g样品。Lys-EGCG共轭物的抗菌和抗氧化活性高于纯EGCG和Lys(P<0.05)。用EGCG对Lys进行化学修饰可得到具有双重功能(抗菌/抗氧化)的生物共轭物,拓宽了Lys和EGCG在食品、化妆品和制药等不同领域的应用范围。

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