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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2022, Vol. 48 Issue (3): 377-382    DOI: 10.3785/j.issn.1008-9209.2021.06.211
动物科学与动物医学     
铁饱和度对乳铁蛋白抑菌活性的影响
王振杰(),张康,梁莉,熊晴晴,杜华华()
浙江大学饲料科学研究所，浙江省饲料与动物营养重点实验室，杭州 310058
Effects of iron saturation on antibacterial activity of lactoferrin
Zhenjie WANG(),Kang ZHANG,Li LIANG,Qingqing XIONG,Huahua DU()
Key Laboratory of Feed and Animal Nutrition of Zhejiang Province, Institute of Feed Sciences, Zhejiang University, Hangzhou 310058, China
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摘要：

本试验通过制备不同铁饱和度的猪乳铁蛋白（lactoferrin, Lf），探讨其对细菌生长的影响。根据铁饱和度将试验分为3组，对照组（不添加任何Lf）、铁不饱和乳铁蛋白处理组（Apo-Lf，铁饱和度为6.9%）和铁饱和乳铁蛋白处理组（Holo-Lf，铁饱和度为100.0%），分别处理大肠埃希菌K88、金黄色葡萄球菌和鼠伤寒沙门菌。结果表明：1）天然Lf和Apo-Lf能显著抑制大肠埃希菌K88和金黄色葡萄球菌的生长，而Holo-Lf没有表现出抑菌活性。2）补铁可以消除Apo-Lf对大肠埃希菌K88的抑制作用，说明Apo-Lf是通过螯合铁发挥抑菌功能的。3）扫描电镜观察结果显示，Apo-Lf破坏了大肠埃希菌K88和金黄色葡萄球菌的细菌膜结构，但Holo-Lf对所有被测细菌都没有影响。4）三维结构预测结果提示，Apo-pLf和Holo-pLf的三维结构开放程度明显不同，Apo-pLf的活性位点可能更容易与细菌直接作用。综上所述，猪乳铁蛋白的抑菌活性依赖于其铁饱和度，Apo-Lf既可以通过螯合铁的方式抑制细菌生长，也可以通过破坏细菌膜结构方式抑制大肠埃希菌K88和金黄色葡萄球菌的生长。
关键词： 乳铁蛋白;  抑菌;  铁饱和度;  铁不饱和乳铁蛋白;  铁饱和乳铁蛋白    
Abstract:

The purpose of this study was to elucidate how iron-depleted and iron-saturated forms of lactoferrin (Lf) exert its antibacterial activity in vitro. Bacteria were divided into three groups by treated with different forms of porcine Lf: the control group (without addition of Lf), apolactoferrin (Apo-Lf, iron saturation is 6.9%) group and hololactoferrin (Holo-Lf, iron saturation is 100.0%) group. The results showed that: 1) Native Lf and Apo-Lf significantly inhibited the growth of Escherichia coli K88 and Staphylococcus aureus (P＜0.01), which was not observed in Holo-Lf group. 2) Apo-Lf exerted antibacterial effect by chelating iron, and iron supplementation could eliminate the inhibitory effect of Apo-Lf on E. coli K88. 3) Scanning electron microscope (SEM) results revealed that Apo-Lf damaged the surface membranes of E. coli K88 and S. Aureus. However, Holo-Lf showed no effect on all of tested bacteria. 4) Predicted three-dimensional structures showed that the structures of Apo-Lf and Holo-Lf were markedly different, and the active site of Apo-Lf was more likely to directly interact with bacteria. Taken together, Apo-Lf inhibited the growth of E. coli K88 and S. Aureus by chelating iron or destroying the surface of bacteria.
Key words: lactoferrin    antibacterial    iron saturation    apolactoferrin    hololactoferrin
收稿日期: 2021-06-21 出版日期: 2022-07-07
CLC:  S 816.7  
基金资助: 浙江省自然科学基金项目(LZ20C170004);国家自然科学基金项目(31872363)
通讯作者: 杜华华     E-mail: 1035043032@qq.com;huahuadu@zju.edu.cn
作者简介: 王振杰（https://orcid.org/0000-0003-1718-5649），E-mail：1035043032@qq.com
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引用本文:

王振杰,张康,梁莉,熊晴晴,杜华华. 铁饱和度对乳铁蛋白抑菌活性的影响[J]. 浙江大学学报（农业与生命科学版）, 2022, 48(3): 377-382.

Zhenjie WANG,Kang ZHANG,Li LIANG,Qingqing XIONG,Huahua DU. Effects of iron saturation on antibacterial activity of lactoferrin. Journal of Zhejiang University (Agriculture and Life Sciences), 2022, 48(3): 377-382.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2021.06.211    或    https://www.zjujournals.com/agr/CN/Y2022/V48/I3/377

图1  不同浓度乳铁蛋白对大肠埃希菌K88生长的影响**表示在P＜0.01水平差异有高度统计学意义。
图2  Apo-Lf和Holo-Lf对细菌生长的影响
图3  补铁对Lf处理的大肠埃希菌K88生长的影响
图4  培养基中的铁对细菌生长的影响
图5  50.0 µg/mL Apo-Lf和50.0 µg/mL Holo-Lf对细菌形态的影响红色箭头表示细菌受损，形态破损。
图6  Apo-Lf和Holo-Lf的三维结构预测红色箭头表示铁结合部位。
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