乳铁蛋白抗菌机理研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2017-0354

摘要/Abstract

摘要：

乳铁蛋白（Lactoferrin,LF）是发现于哺乳动物初乳中具有多种生物活性的单体糖蛋白,对幼体初期免疫具有重要作用。前人研究发现LF蛋白的酶解后其抗菌活性得到增强,是由于酶解后产生了比其本身抗菌能力更强的肽段。目前关于LF蛋白抗菌活性的报道很多,但其抗菌的具体分子机制尚不清楚。以LF蛋白与细菌细胞膜的结合能力和LF蛋白酶解产物的抗菌活性为切入点,揭示了LF蛋白行使抗菌功能的过程,认为LF蛋白与细菌表面相结合而发生结构变化,进而暴露出敏感的酶切位点,经酶解而释放抗菌活性肽,这些抗菌肽破坏细菌的细胞膜结构,达到抑菌或杀菌的目的。对LF蛋白抗菌机理的阐释将为研制抗菌能力更强的活性蛋白提供理论依据。

关键词: 乳铁蛋白, 抗菌活性, 抗菌肽, 膜结合

Abstract:

Lactoferrin（LF）is a monomer glycoprotein in mammalian colostrum with multi biological activities,thus it plays a vital role in initial immunologic reconstitution for young mammals. Previous studies have discovered that enzymatic hydrolysates of lactoferrin have enhanced antibacterial activity compared to lactoferrin because more active antibacterial peptides were produced by the enzymolysis. At present,molecular mechanism of antibacterial function of lactoferrin is still unclear although many antibacterial activities of lactoferrin were reported. In this paper,combination of the bacterial cell membranes and the antibacterial activity of enzymatic hydrolysates were taken as the breakthrough point,and the process of LF acting in antibacterial function was explored. It was considered that the mechanism of the antibacterial function of lactoferrin was that the structure of lactoferrin changed following binding to bacterial surface,then the sensitive restriction enzyme cutting sites of lactoferrin were exposed,and many antibacterial peptides were released after enzymolysis；subsequently the peptides destroyed the membrane structures of bacterial cells,and thus the purpose of the bacteriostasis or sterilization achieved. In summary,the elucidation of antibacterial mechanism of lactoferrin provides the theoretical basis for developing active proteins with stronger antibacterial ability.

Key words: lactoferrin, antibacterial activity, antimicrobial peptide, membrane binding

裴杰,褚敏,包鹏甲,阎萍,郭宪. 乳铁蛋白抗菌机理研究进展[J]. 生物技术通报, 2017, 33(9): 56-63.

PEI Jie,CHU Min,BAO Peng-jia,YAN Ping,GUO Xian. Research Progress on Antibacterial Mechanism of Lactoferrin[J]. Biotechnology Bulletin, 2017, 33(9): 56-63.

参考文献

[1] Wakabayashi H, Yamauchi K, Takase M. Lactoferrin research, technology and applications[J] . International Dairy Journal, 2006, 16（11）：1241-1251.
[2] 朱艳萍, 滕达, 田子罡, 等. 乳铁蛋白分子结构及其抗菌机制[J] . 生物技术通报, 2010（6）：37-42.
[3] Lambert LA. Molecular evolution of the transferrin family and associated receptors[J] . Biochimica Et Biophysica Acta-General Subjects, 2012, 1820（3）：244-255.
[4] Duarte DC, Nicolau A, Teixeira JA, et al. The effect of bovine milk lactoferrin on human breast cancer cell lines[J] . Journal of Dairy Science, 2011, 94（1）：66-76.
[5] Pierce A, Legrand D. Advances in lactoferrin research Introduction [J] . Biochimie, 2009, 91（1）：1-2.
[6] Ochoa TJ, Sizonenko SV. Lactoferrin and prematurity：a promising milk protein?[J] . Biochemistry and Cell Biology, 2017, 95（1）：22-30.
[7] Wang X, Wang X, Hao Y, et al. Research and development on lactoferrin and its derivatives in China from 2011-2015[J] . Biochemistry and Cell Biology, 2017, 95（1）：162-170.
[8] Moore SA, Anderson BF, Groom CR, et al. Three-dimensional struc-ture of diferric bovine lactoferrin at 2.8 angstrom resolution[J] . Journal of Molecular Biology, 1997, 274（2）：222-236.
[9] Rikova K, Guo A, Zeng Q, et al. Global survey of phosphotyrosine signaling identifies oncogenic kinases in lung cancer[J] . Cell, 2007, 131（6）：1190-1203.
[10] Barboza M, Pinzon J, Wickramasinghe S, et al. Glycosylation of human milk lactoferrin exhibits dynamic changes during early lactation enhancing its role in pathogenic bacteria-host interactions[J] . Molecular & Cellular Proteomics, 2012, 11（6）.
[11] Mayeur S, Spahis S, Pouliot Y, et al. Lactoferrin, a Pleiotropic Protein in Health and Disease[J] . Antioxidants & Redox Signaling, 2016, 24（14）：813-835.
[12] Valenti P, Antonini G. Lactoferrin：an important host defence against microbial and viral attack[J] . Cellular and Molecular Life Sciences, 2005, 62（22）：2576-2587.
[13] Del Olmo A, Calzada J, Nunez M. Effect of lactoferrin and its derivatives against gram-positive bacteria in vitro and, combined with high pressure, in chicken breast fillets[J] . Meat Science, 2012, 90（1）：71-76.
[14] 文鹏程, 余丹丹, 汪昕昕, 等. 不同处理条件对乳铁蛋白构象的影响研究[J] . 光谱学与光谱分析, 2012（1）：162-165.
[15] Zhang J, Li L, Cai Y, et al. Expression of active recombinant human lactoferrin in the milk of transgenic goats[J] . Protein Expression and Purification, 2008, 57（2）：127-135.
[16] Jenssen H, Hancock REW. Antimicrobial properties of lactoferrin[J] . Biochimie, 2009, 91（1）：19-29.
[17] Gonzalez-Chavez SA, Arevalo-Gallegos S, Rascon-Cruz Q. Lactofe-rrin：structure, function and applications[J] . International Journal of Antimicrobial Agents, 2009, 33（4）：301-306.
[18] Yen C-C, Shen C-J, Hsu W-H, et al. Lactoferrin：an iron-binding antimicrobial protein against Escherichia coli infection[J] . Biometals, 2011, 24（4）：585-594.
[19] Leon-Sicairos N, Canizalez-Roman A, de la Garza M, et al. Bacteri-cidal effect of lactoferrin and lactoferrin chimera against halophilic Vibrio parahaemolyticus[J] . Biochimie, 2009, 91（1）：133-140.
[20] Mela I, Aumaitre E, Williamson A-M, et al. Charge reversal by salt-induced aggregation in aqueous lactoferrin solutions[J] . Colloids and Surfaces B-Biointerfaces, 2010, 78（1）：53-60.
[21] Rahman MM, Kim W-S, Ito T, et al. Growth promotion and cell bin-ding ability of bovine lactoferrin to Bifidobacterium longum[J] . Anaerobe, 2009, 15（4）：133-137.
[22] 张伟, 任发政, 葛绍阳, 等. 拉曼光谱研究乳铁蛋白及其肽段与DPPC、DPPG脂质体的相互作用[J] . 光谱学与光谱分析, 2011（6）：1533-1536.
[23] Wakabayashi H, Yamauchi K, Kobayashi T, et al. Inhibitory effects of lactoferrin on growth and biofilm formation of Porphyromonas gingivalis and Prevotella intermedia[J] . Antimicrobial Agents and Chemotherapy, 2009, 53（8）：3308-3316.
[24] Ammons MCB, Ward LS, Dowd S, et al. Combined treatment of Pseudomonas aeruginosa biofilm with lactoferrin and xylitol inhibits the ability of bacteria to respond to damage resulting from lactoferrin iron chelation[J] . International Journal of Antimicrobial Agents, 2011, 37（4）：316-323.
[25] Ammons MCB, Ward LS, Fisher ST, et al. In vitro susceptibility of established biofilms composed of a clinical wound isolate of Pseudomonas aeruginosa treated with lactoferrin and xylitol[J] . International Journal of Antimicrobial Agents, 2009, 33（3）：230-236.
[26] Ellison RT, III, Giehl TJ, LaForce FM. Damage of the outer membrane of enteric gram-negative bacteria by lactoferrin and transferrin[J] . Infection and Immunity, 1988, 56（11）：2774-2781.
[27] Caccavo D, Afeltra A, Pece S, et al. Lactoferrin-lipid A-lipopolysaccharide interaction：Inhibition by anti-human lactoferrin monoclonal antibody AGM 10. 14[J] . Infection and Immunity, 1999, 67（9）：4668-4672.
[28] Japelj B, Pristovsek P, Majerle A, et al. Structural origin of endo-toxin neutralization and antimicrobial activity of a lactoferrin-based peptide[J] . Journal of Biological Chemistry, 2005, 280（17）：16955-16961.
[29] Kim WS, Rahman MM, Shimazaki KI. Antibacterial activity and binding ability of bovine lactoferrin against Pseudomonas spp. [J] . Journal of Food Safety, 2008, 28（1）：23-33.
[30] Andre GO, Politano WR, Mirza S, et al. Combined effects of lactoferrin and lysozyme on Streptococcus pneumoniae killing[J] . Microbial Pathogenesis, 2015, 89：7-17.
[31] Senkovich O, Cook WJ, Mirza S, et al. Structure of a complex of human lactoferrin N-lobe with pneumococcal surface protein A provides insight into microbial defense mechanism[J] . Journal of Molecular Biology, 2007, 370（4）：701-713.
[32] Samaniego-Barron L, Luna-Castro S, Pina-Vazquez C, et al. Two outer membrane proteins are bovine lactoferrin-binding proteins in Mannheimia haemolytica A1[J] . Veterinary Research, 2016, 47.
[33] Wong H, Schryvers AB. Bacterial lactoferrin-binding protein A binds to both domains of the human lactoferrin C-lobe[J] . Microbiology-Sgm, 2003, 149：1729-1737.
[34] Ostan N, Morgenthau A, Yu RH, et al. A comparative, cross-species investigation of the properties and roles of transferrin-and lactoferrin-binding protein B from pathogenic bacteria[J] . Biochemistry and Cell Biology, 2017, 95（1）：5-11.
[35] Zinger-Yosovich KD, Sudakevitz D, Iluz D, et al. Analyses of diverse mammals’ milk and lactoferrin glycans using five pathogenic bacterial lectins[J] . Food Chemistry, 2011, 124（4）：1335-1342.
[36] Yu T, Guo C, Wang J, et al. Comprehensive characterization of the site-specific N-glycosylation of wild-type and recombinant human lactoferrin expressed in the milk of transgenic cloned cattle[J] . Glycobiology, 2011, 21（2）：206-224.
[37] Del Olmo A, Calzada J, Nunez M. Short communication：Antimic-robial effect of lactoferrin and its amidated and pepsin-digested derivatives against Salmonella Enteritidis and Pseudomonas fluore-scens[J] . Journal of Dairy Science, 2010, 93（9）：3965-3969.
[38] Baker EN, Baker HM. A structural framework for understanding the multifunctional character of lactoferrin[J] . Biochimie, 2009, 91（1）：3-10.
[39] Morgenthau A, Pogoutse A, Adamiak P, et al. Bacterial receptors for host transferrin and lactoferrin：molecular mechanisms and role in host-microbe interactions[J] . Future Microbiology, 2013, 8（12）：1575-1585.
[40] Le Parc A, Karav S, Rouquie C, et al. Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows[J] . Plos One, 2017, 12（2）.
[41] Sojar HT, Hamada N, Genco RJ. Structures involved in the interaction of Porphyromonas gingivalis fimbriae and human lactoferrin[J] . Febs Letters, 1998, 422（2）：205-208.
[42] Kamiya H, Ehara T, Matsumoto T. Inhibitory effects of lactoferrin on biofilm formation in clinical isolates of Pseudomonas aeruginosa[J] . Journal of Infection and Chemotherapy, 2012, 18（1）：47-52.
[43] Diarra MS, Lacasse P, Deschenes E, et al. Ultrastructural and cytochemical study of cell wall modification by lactoferrin, lactoferricin and penicillin G against Staphylococcus aureus[J] . Journal of Electron Microscopy, 2003, 52（2）：207-215.
[44] Quintieri L, Caputo L, Monaci L, et al. Antimicrobial efficacy of pepsin-digested bovine lactoferrin on spoilage bacteria contaminating traditional Mozzarella cheese[J] . Food Microbiology, 2012, 31（1）：64-71.
[45] Nibbering PH, Ravensbergen E, Welling MM, et al. Human lactoferrin and peptides derived from its N terminus are highly effective against infections with antibiotic-resistant bacteria[J] . Infection and Immunity, 2001, 69（3）：1469-1476.
[46] Tomita M, Wakabayashi H, Shin K, et al. Twenty-five years of research on bovine lactoferrin applications[J] . Biochimie, 2009, 91（1）：52-57.
[47] Duchardt F, Ruttekolk IR, Verdurmen WPR, et al. A cell-penetrating peptide derived from human lactoferrin with conformation-dependent uptake efficiency[J] . Journal of Biological Chemistry, 2009, 284（52）：36099-36108.
[48] Moniruzzaman M, Alam JM, Dohra H, et al. Antimicrobial peptide lactoferricin B-induced rapid leakage of internal contents from single giant unilamellar vesicles[J] . Biochemistry, 2015, 54（38）：5802-5814.
[49] Hwang PM, Zhou N, Shan X, et al. Three-dimensional solution structure of lactoferricin B, an antimicrobial peptide derived from bovine lactoferrin[J] . Biochemistry, 1998, 37（12）：4288-4298.
[50] Ghosh A, Datta A, Jana J, et al. Sequence context induced antimi-crobial activity：insight into lipopolysaccharide permeabilization[J] . Molecular Biosystems, 2014, 10（6）：1596-1612.
[51] van der Kraan MIA, Groenink J, Nazmi K, et al. Lactoferrampin：a novel antimicrobial peptide in the N1-domain of bovine lactoferrin[J] . Peptides, 2004, 25（2）：177-183.
[52] Haney EF, Lau F, Vogel HJ. Solution structures and model membrane interactions of lactoferrampin, an antimicrobial peptide derived from bovine lactoferrin[J] . Biochimica Et Biophysica Acta-Biomembranes, 2007, 1768（10）：2355-2364.
[53] Haney EF, Nazmi K, Lau F, et al. Novel lactoferrampin antimicrobial peptides derived from human lactoferrin[J] . Biochimie, 2009, 91（1）：141-154.
[54] Bolscher JGM, Adao R, Nazmi K, et al. Bactericidal activity of Lfchimera is stronger and less sensitive to ionic strength than its constituent lactoferricin and lactoferrampin peptides[J] . Biochimie, 2009, 91（1）：123-132.
[55] Haney EF, Nazmi K, Bolscher JGM, et al. Structural and biophysical characterization of an antimicrobial peptide chimera comprised of lactoferricin and lactoferrampin[J] . Biochimica Et Biophysica Acta-Biomembranes, 2012, 1818（3）：762-775.
[56] Bjorn C, Mahlapuu M, Mattsby-Baltzer I, et al. Anti-infective efficacy of the lactoferrin-derived antimicrobial peptide HLR1r[J] . Peptides, 2016, 81：21-28.
[57] Zweytick D, Deutsch G, Andra J, et al. Studies on lactoferricin-derived Escherichia coli membrane-active peptides reveal differences in the mechanism of N-acylated versus nonacylated peptides[J] . Journal of Biological Chemistry, 2011, 286（24）：21266-21276.
[58] Mishra B, Leishangthem GD, Gill K, et al. A novel antimicrobial peptide derived from modified N-terminal domain of bovine lactoferrin：Design, synthesis, activity against multidrug-resistant bacteria and Candida[J] . Biochimica Et Biophysica Acta-Biomembranes, 2013, 1828（2）：677-686.
[59] Viejo-Diaz M, Andres MT, Fierro JF. Different anti-Candida activities of two human lactoferrin-derived peptides, Lfpep and kaliocin-1[J] . Antimicrobial Agents and Chemotherapy, 2005, 49（7）：2583-2588.
[60] Morici P, Fais R, Rizzato C, et al. Inhibition of Candida albicans biofilm formation by the synthetic lactoferricin derived peptide hLF1-11[J] . PLoS One, 2016, 11（11）.
[61] Kondori N, Baltzer L, Dolphin GT, et al. Fungicidal activity of human lactoferrin-derived peptides based on the antimicrobial alpha beta region[J] . International Journal of Antimicrobial Agents, 2011, 37（1）：51-57.