Research Center for Biotechnology, Indonesian Institute of Science (LIPI), Bogor, Indonesia.
School of Biotechnology, Bogor Agricultural University, Bogor, Indonesia.
Mol Biol Rep. 2019 Dec;46(6):6501-6512. doi: 10.1007/s11033-019-05096-9. Epub 2019 Oct 3.
Enteropathogenic Escherichia coli (EPEC) is one of the resistance bacteria towards antibiotics and have been raising problem during treatments. Therefore, a new antibiotic candidate is required. Plantaricin E and F recombinant have been successfully produced by a GRAS host Lactococcus lactis. This study was aimed to evaluate the efficacy and toxicity of plantaricin E and F recombinant against EPEC K1.1 infection by in vivo assay. The production of plantaricin E and F recombinants from Lactococcus lactis was conducted and encapsulated. The in vivo study was carried out by inoculating the mice perorally with EPEC K1.1 for 7 days then treated with 100, 250, and 500 mg/kg body weight/day of recombinant plantaricin E and F for another 7 days. The toxicity assay were observed in ddY mice using various concentrations of treatment (50, 100, 1000, and 5000 mg/kg/body weight) doses perorally for 48 h. The result showed that the plantaricin E and F recombinant were successfully produced in Lactococcus lactis expression host with 3.7 kDa and 3.8 kDa in size. The efficacy study revealed the optimal doses of plantaricin E and F recombinant against EPEC K1.1 infection was 250 mg/kgBW for plantaricin E and 500 mg/kgBW for plantaricin F. The plantarisin E and F recombinant treatment showed improvement in leukocyte, hematocrit, and hemoglobin levels as well in decreasing malondialdehyde (MDA) level. Observation of the intestine histopathology showed small amounts of mononuclear inflammatory cell infiltration than the other groups of treatment. The acute toxicity assay showed that there was no mortality observed during the assay, even after 5000 mg/kg body weight of plantarisin E and F recombinant treatment (LD > 5000 mg/KgBW). The hematological and biochemical observations showed normal levels in leukocytes, erythrocytes, hematocrit, hemoglobin, platelets, urea, creatinine, and alanine transaminase aspartate transaminase (SGOT and SGPT) while histopathological observation shows a picture of normal liver and kidney cells. This study confirmed the application of bacteriocin for further academic and industrial purposes as a non-toxic substance for food preservative and antibiotic candidate.
肠致病性大肠杆菌（EPEC）是一种对抗生素具有耐药性的细菌，在治疗过程中一直存在问题。因此，需要一种新的抗生素候选物。植物乳杆菌（Lactococcus lactis）GRAS 宿主已成功生产出重组植物杀菌素 E 和 F。本研究旨在通过体内试验评估重组植物杀菌素 E 和 F 对 EPEC K1.1 感染的疗效和毒性。通过发酵和包封生产植物杀菌素 E 和 F 的重组体。通过口接种 EPEC K1.1 对小鼠进行为期 7 天的体内研究，然后用 100、250 和 500mg/kg 体重/天的重组植物杀菌素 E 和 F 治疗 7 天。在 ddY 小鼠中观察毒性试验，使用各种浓度的处理（50、100、1000 和 5000mg/kg/体重）经口给药 48 小时。结果表明，重组植物杀菌素 E 和 F 在 Lactococcus lactis 表达宿主中成功表达，大小分别为 3.7kDa 和 3.8kDa。疗效研究表明，重组植物杀菌素 E 和 F 对 EPEC K1.1 感染的最佳剂量为 250mg/kgBW 用于植物杀菌素 E 和 500mg/kgBW 用于植物杀菌素 F。植物杀菌素 E 和 F 重组体治疗可改善白细胞、红细胞压积和血红蛋白水平，并降低丙二醛（MDA）水平。观察肠组织病理学显示，单核炎性细胞浸润量比其他治疗组少。急性毒性试验表明，即使在给予 5000mg/kg 体重的植物杀菌素 E 和 F 重组体（LD>5000mg/kgBW）后，试验过程中也没有观察到死亡。血液学和生化学观察显示白细胞、红细胞、红细胞压积、血红蛋白、血小板、尿素、肌酐和丙氨酸氨基转移酶天门冬氨酸氨基转移酶（SGOT 和 SGPT）水平正常，组织病理学观察显示肝和肾细胞正常。本研究证实了细菌素在食品防腐剂和抗生素候选物等学术和工业领域的应用，作为一种无毒物质。
