Potential of the virion-associated peptidoglycan hydrolase HydH5 and its derivative fusion proteins in milk biopreservation.
Potential of the virion-associated peptidoglycan hydrolase HydH5 and its derivative fusion proteins in milk biopreservation.
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DOI：
10.1371/journal.pone.0054828
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Rodríguez A
中科院分区：
文献类型：
--
作者：
Rodríguez-Rubio L;Martínez B;Donovan DM;García P;Rodríguez A
噬菌体裂解酶作为一种新型的抗革兰氏阳性菌的抗菌剂，近年来引起了人们的极大兴趣。在这项工作中，HydH 5 [由金黄色葡萄球菌噬菌体vB_SauS-phiIPLA 88编码的病毒体相关肽聚糖水解酶（VAPGH）]和HydH 5与溶葡萄球菌酶之间产生的三种不同融合蛋白在牛奶中的抗微生物活性已经被评估。在104 CFU/mL菌株S的挑战试验中，使用市售延长货架期全脂牛奶（ESL）确认了5种蛋白质（HydH 5、HydH 5 Lyso、HydH 5 SH 3b、CHAPSH 3b和溶葡萄球菌酶）的裂解活性。金黄色葡萄球菌Sa 9. HydH 5、HydH 5Lyso和HydH 5SH 3b（3.5 µM）在37 ℃下保持葡萄球菌活菌计数低于对照培养物6 h。在加入溶解酶后仅15分钟，效果就很明显。值得注意的是，溶葡萄球菌酶和CHAPSH 3b显示出最高的葡萄球菌溶解保护，因为它们能够在37°C下以较低浓度（1 μM）分别在添加后立即或15分钟根除初始葡萄球菌攻击。CHAPSH 3b在室温下显示出相同的抗葡萄球菌作用（1.65 µM）。在实验的剩余时间（长达6小时）内未观察到再生长。还在生乳（全脂和脱脂）和巴氏灭菌乳（全脂和脱脂）中测定了CHAPSH 3b活性（1.65 µM）。在两种温度下，牛奶的巴氏灭菌明显增强了全脂牛奶和脱脂牛奶中CHAPSH 3b的葡萄球菌溶解活性。这种效果在室温下最显著，因为这种蛋白质能够减少S。加入后立即将金黄色葡萄球菌活菌计数降至不可检测的水平，在实验期间（360分钟）未检测到再生长。此外，已知CHAPSH 3b蛋白是耐热的，并且在巴氏灭菌处理后和在4°C下储存3天后保留一些裂解活性。这些结果可能有助于使用肽聚糖水解酶HydH 5及其衍生物融合体，特别是CHAPSH 3b，作为控制乳制品中不良细菌的生物控制剂。
Bacteriophage lytic enzymes have recently attracted considerable interest as novel antimicrobials against Gram-positive bacteria. In this work, antimicrobial activity in milk of HydH5 [a virion-associated peptidoglycan hydrolase (VAPGH) encoded by the Staphylococcus aureus bacteriophage vB_SauS-phiIPLA88], and three different fusion proteins created between HydH5 and lysostaphin has been assessed. The lytic activity of the five proteins (HydH5, HydH5Lyso, HydH5SH3b, CHAPSH3b and lysostaphin) was confirmed using commercial whole extended shelf-life milk (ESL) in challenge assays with 104 CFU/mL of the strain S. aureus Sa9. HydH5, HydH5Lyso and HydH5SH3b (3.5 µM) kept the staphylococcal viable counts below the control cultures for 6 h at 37°C. The effect is apparent just 15 minutes after the addition of the lytic enzyme. Of note, lysostaphin and CHAPSH3b showed the highest staphylolytic protection as they were able to eradicate the initial staphylococcal challenge immediately or 15 min after addition, respectively, at lower concentration (1 µM) at 37°C. CHAPSH3b showed the same antistaphyloccal effect at room temperature (1.65 µM). No re-growth was observed for the remainder of the experiment (up to 6 h). CHAPSH3b activity (1.65 µM) was also assayed in raw (whole and skim) and pasteurized (whole and skim) milk. Pasteurization of milk clearly enhanced CHAPSH3b staphylolytic activity in both whole and skim milk at both temperatures. This effect was most dramatic at room temperature as this protein was able to reduce S. aureus viable counts to undetectable levels immediately after addition with no re-growth detected for the duration of the experiment (360 min). Furthermore, CHAPSH3b protein is known to be heat tolerant and retained some lytic activity after pasteurization treatment and after storage at 4°C for 3 days. These results might facilitate the use of the peptidoglycan hydrolase HydH5 and its derivative fusions, particularly CHAPSH3b, as biocontrol agents for controlling undesirable bacteria in dairy products.
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影响因子：
4.1
作者：
Fischetti, Vincent A.
通讯作者：
Fischetti, Vincent A.
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3.5
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影响因子：
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作者：
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