Novel Antibacterial Peptides Isolated from the Maillard Reaction Products of Half-Fin Anchovy (Setipinna taty) Hydrolysates/Glucose and Their Mode of Action in Escherichia Coli
Novel Antibacterial Peptides Isolated from the Maillard Reaction Products of Half-Fin Anchovy (Setipinna taty) Hydrolysates/Glucose and Their Mode of Action in Escherichia Coli
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DOI：
10.3390/md17010047
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发表时间：
2019-01
期刊：
影响因子：
5.4
通讯作者：
Jiaxing Wang;Rong-bian Wei;Ru Song
中科院分区：
文献类型：
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作者：
Jiaxing Wang;Rong-bian Wei;Ru Song
半鳍金枪鱼水解液与葡萄糖的美拉德反应产物（MRPs），命名为HAHp（9.0）-G MRPs，经体积排阻色谱分离后，分为3个主要组分（F1-F3）。F2菌株对大肠杆菌（E. coli）的抗菌活性最强。大肠杆菌（coli），并显示自身产生过氧化氢（H2 O2）。通过反相高效液相色谱法将F2的疏水提取物进一步分离成亚级分HE-F2-1和HE-F2-2。使用液相色谱-电喷雾电离/多级质谱法从HE-F2-1中鉴定出9种肽，从HE-F2-2中鉴定出2种肽。合成了三种肽：FEDQLR（HGM-Hp 1）、ALERTF（HGM-Hp 2）和RHPEYAVSVLLR（HGM-Hp 3），净电荷分别为-1，0和+1。合成肽对大肠杆菌的最低抑菌浓度为2 mg/mL。杆菌与对数生长期的E. coli、HGM-Hp 1和HGM-Hp 2诱导细胞外和细胞内H2 O2生成均显著增加。然而，HGM-Hp 3仅显著增加了E.杆菌E.加入HGM-Hp 1或HGM-Hp 2后的大肠杆菌悬浮液表明通过不可逆的膜损伤破坏细胞完整性。这是首次报道的水解产物MRPs衍生肽，可能通过诱导细胞内H2 O2的产生来执行抗菌活性。
The Maillard reaction products (MRPs) of half-fin anchovy hydrolysates and glucose, named as HAHp(9.0)-G MRPs, were fractionated by size exclusion chromatography into three major fractions (F1–F3). F2, which demonstrated the strongest antibacterial activity against Escherichia coli (E. coli) and showed self-production of hydrogen peroxide (H2O2), was extracted by solid phase extraction. The hydrophobic extract of F2 was further isolated by reverse phase-high performance liquid chromatography into sub-fractions HE-F2-1 and HE-F2-2. Nine peptides were identified from HE-F2-1, and two peptides from HE-F2-2 using liquid chromatography-electrospray ionization/multi-stage mass spectrometry. Three peptides, FEDQLR (HGM-Hp1), ALERTF (HGM-Hp2), and RHPEYAVSVLLR (HGM-Hp3), with net charges of −1, 0, and +1, respectively, were synthesized. The minimal inhibitory concentration of these synthetic peptides was 2 mg/mL against E. coli. Once incubated with logarithmic growth phase of E. coli, HGM-Hp1 and HGM-Hp2 induced significant increases of both extracellular and intracellular H2O2 formation. However, HGM-Hp3 only dramatically enhanced intracellular H2O2 production in E. coli. The increased potassium ions in E. coli suspension after addition of HGM-Hp1 or HGM-Hp2 indicated the destruction of cell integrity via irreversible membrane damage. It is the first report of hydrolysates MRPs-derived peptides that might perform the antibacterial activity via inducing intracellular H2O2 production.