Effects of iron saturation on antibacterial activity of lactoferrin

doi:10.3785/j.issn.1008-9209.2021.06.211

Journal of Zhejiang University (Agriculture and Life Sciences)

2022, Vol. 48

Issue (3): 377-382 DOI: 10.3785/j.issn.1008-9209.2021.06.211

Animal sciences & veterinary medicines

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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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

Received: 21 June 2021 Published: 07 July 2022

CLC:

S 816.7

Corresponding Authors: Huahua DU E-mail: 1035043032@qq.com;huahuadu@zju.edu.cn

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Cite this article:

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.

URL:

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

铁饱和度对乳铁蛋白抑菌活性的影响

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

Fig. 1 Effects of different concentrations of Lf on the growth of E. coli K88Double asterisks (**) indicate highly significant differences at the 0.01 probability level.

Fig. 2 Effects of Apo-Lf and Holo-Lf on the growth of bacteria

Fig. 3 Effects of Fe supplement on the growth of Lf-treated E. coli K88

Fig. 4 Effects of Fe in the culture medium on the growth of bacteria

Fig. 5 Effects of 50.0 µg/mL Apo-Lf and 50.0 µg/mL Holo-Lf on the morphologies of bacteriaThe damaged bacterial morphology are indicated by red arrows.

Fig. 6 Predicted three-dimensional structures of Apo-Lf and Holo-LfIron binding sites are indicated by red arrows.


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