Preparation and characterization of endolysin-containing liposomes and evaluation of their antimicrobial activities against gram-negative bacteria
Preparation and characterization of endolysin-containing liposomes and evaluation of their antimicrobial activities against gram-negative bacteria
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DOI：
10.1016/j.enzmictec.2019.05.006
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发表时间：
2019-09-01
影响因子：
3.4
通讯作者：
Ryu, Sangryeol
中科院分区：
文献类型：
--
作者：
Bai, Jaewoo;Yang, Eunhye;Ryu, Sangryeol
抗生素在治疗细菌感染中的过度使用和滥用导致耐药细菌的迅速出现，这表明迫切需要开发控制耐药细菌的替代策略。内溶素是噬菌体编码的酶，可以降解细菌细胞壁中的肽聚糖，它们作为替代抗菌剂具有很大的潜力。然而，重组内溶素的外源性应用仅限于革兰氏阳性菌，因为内溶素不能穿透革兰氏阴性菌的外膜。在这里，开发了脂质体介导的内溶素包封系统，并测试了其穿透革兰氏阴性菌外膜的能力。分离、表征噬菌体来源的内溶素BSP 16 Lys，并将其封装到由二棕榈酰磷脂酰胆碱（DPPC）、胆固醇和十六烷基胺组成的阳离子脂质体中。BSP 16 Lys包封的脂质体具有高的zeta电位值（超过30 mV），平均直径为303 nm。BSP 16 Lys在脂质体中的包封率为35.27%。用BSP 16 Lys包封的脂质体处理的鼠伤寒沙门氏菌和大肠杆菌细胞分别显示出2.2-log CFU/mL和1.6-log CFU/mL的活细胞数减少，而不处理膜透化剂。这些结果显示了脂质体介导的内溶素递送用于针对革兰氏阴性菌的外源性应用的潜力。
The overuse and misuse of antibiotics in treating bacterial infections cause the rapid emergence of drug-resistant bacteria, suggesting that the development of alternative strategies to control antibiotic-resistant bacteria is urgently needed. Endolysins are bacteriophage-encoded enzymes that can degrade peptidoglycan in bacterial cell walls, and they have great potential as alternative antimicrobial agents. However, exogenous application of recombinant endolysin is limited to Gram-positive bacteria because endolysins cannot penetrate the outer membrane of Gram-negative bacteria. Here, a liposome-mediated endolysin encapsulation system was developed, and its ability to penetrate the outer membrane of Gram-negative bacteria was tested. The phage-derived endolysin BSP16Lys was isolated, characterized, and used for encapsulation into a cationic liposome comprised of dipalmitoylphosphatidylcholine (DPPC), cholesterol, and hexadecylamine. The BSP16Lys-encapsulated liposome had a high zeta potential value (over 30 mV) with an average diameter of 303 nm. The encapsulation efficiency of BSP16Lys into the liposome was 35.27%. Salmonella Typhimuriumand Escherichia coli cells treated with BSP16Lys-encapsulated liposomes showed 2.2-log CFU/mL and 1.6-log CFU/mL reductions in the viable cell numbers, respectively, without treatment of a membrane permeabilizer. These results showed potential for liposome-mediated delivery of endolysin for exogenous application against Gram-negative bacteria.