Analysis of the antibacterial and anti-inflammatory functions of bovine lactoferrin functional fragment expressed in Escherichia coli via engineering

IF 1.8 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2026-03-01 Epub Date: 2025-11-29 DOI:10.1016/j.pep.2025.106865

Hui Teng, Zizi Ding, Yujie Wang, Xueliang Liu, Xiangshan Tang, Xuewen Zhang

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Abstract

The bovine lactoferrin is a natural iron-binding glycoprotein that has attracted widespread attention due to its functions such as antibacterial, antiviral, anti-tumor, anti-inflammatory and immunomodulatory effects. In this study, the lactoferrin functional fragment (BlfFf) was expressed in Escherichia coli by genetic engineering expression. The expressed BlfFf mainly existed as inclusion bodies in the host cells. After collecting the inclusion bodies, the BlfFf purified polypeptide was obtained through dissolution, renaturation and further chromatography preparation. The antibacterial test of the purified BlfFf indicated that BlfFf did not exhibit antibacterial activity before pepsin hydrolysis. After hydrolysis by pepsin, BlfFf has obvious antibacterial properties against both Staphylococcus aureus and Escherichia coli represent Gram-positive and Gram-negative bacteria each. Cell experiments have demonstrated that the prepared BlfFf can significantly reduce the expression of inflammation-related genes in the cell model of LPS-stimulated macrophages (Raw264.7) with inflammation model, showing a significant inflammatory inhibitory effect. In Raw264.7 cells treated with BlfFf, the expression levels of various pro-inflammatory cytokines IL-1β, TNF-α, IL-6 and iNOS were significantly decreased. Lower concentrations of BlfFf show a certain cell-promoting growth effect. This experiment demonstrated the multiple biological activities and functions of bacterial genetic engineering in expressing bovine lactoferrin peptides.

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大肠杆菌表达的牛乳铁蛋白功能片段抑菌抗炎功能的工程分析。

牛乳铁蛋白是一种天然的铁结合糖蛋白，因其具有抗菌、抗病毒、抗肿瘤、抗炎和免疫调节等功能而受到广泛关注。本研究利用基因工程技术在大肠杆菌中表达乳铁蛋白功能片段（BlfFf）。表达的BlfFf主要以包涵体形式存在于宿主细胞中。收集包涵体后，通过溶出、复性和进一步层析制备得到BlfFf纯化多肽。对纯化的BlfFf进行抑菌实验表明，在胃蛋白酶水解前BlfFf不表现出抑菌活性。经胃蛋白酶水解后，BlfFf对代表革兰氏阳性菌和革兰氏阴性菌的金黄色葡萄球菌和大肠杆菌均有明显的抑菌作用。细胞实验表明，制备的BlfFf可以显著降低lps刺激的巨噬细胞（Raw264.7）炎症模型中炎症相关基因的表达，显示出明显的炎症抑制作用。在经BlfFf处理的Raw264.7细胞中，各种促炎细胞因子IL-1β、TNF-α、IL-6和iNOS的表达水平均显著降低。低浓度BlfFf具有一定的促细胞生长作用。本实验验证了细菌基因工程表达牛乳铁蛋白多肽的多种生物学活性和功能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.