Peng Zhong, Wang Shaohui, Gide Mussie, Zhu Duolong, Lamabadu Warnakulasuriya Patabendige Hiran Malinda, Li Chunhui, Cai Jianfeng, Sun Xingmin
Department of Molecular Medicine, Morsani College of Medicine, University of South Florida, Tampa, FL, United States.
State Key Laboratory of Agricultural Microbiology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Front Microbiol. 2019 Jan 11;9:3234. doi: 10.3389/fmicb.2018.03234. eCollection 2018.
is the leading cause of worldwide antibiotics-associated diarrhea. In this study, we report the construction and evaluation of a novel bacteriophage lysin-human defensin fusion protein targeting . The fusion protein, designated LHD, is composed of two parts connected by a 3-repeating unit linker "(GGGGS)": the catalytic domain of a lysin protein from a bacteriophage phiC2 (LCD), and the functional domain of a human defensin protein HD. Lytic assays showed that LHD protein had a potent lytic activity against different types of clinical strains, including the epidemic 027, 078, 012, and 087 strains. The minimum inhibitory concentration (MIC) of LHD was 0.78 μg/ml, which was lower than the MIC of the protein LCD (1.56 μg/ml), and the MICs of metronidazole (4 μg/ml) and vancomycin (4 μg/ml). In addition, the LHD protein could lyse strains in different pHs (6.0, 7.0, and 8.0). Evaluation of LHD potency using mouse model of infection (CDI) showed that administration of the LHD protein (twice daily for 7 days) was effective in mitigating the symptoms and reducing the death from CDI. Treatment with LHD also significantly decreased the number of spores and the toxin level in feces from the infected mice. Our data suggest that this novel lysin-human defensin fusion protein has a potential on CDI control.
是全球抗生素相关性腹泻的主要原因。在本研究中，我们报告了一种新型噬菌体溶菌酶-人防御素融合蛋白靶向……的构建及评估。该融合蛋白命名为LHD，由通过3个重复单元连接子“(GGGGS)”连接的两部分组成：来自噬菌体phiC2的溶菌酶蛋白的催化结构域（LCD）和人防御素蛋白HD的功能结构域。溶菌试验表明，LHD蛋白对不同类型的临床……菌株具有强大的溶菌活性，包括流行的027、078、012和087菌株。LHD的最低抑菌浓度（MIC）为0.78μg/ml，低于蛋白LCD的MIC（1.56μg/ml）以及甲硝唑（4μg/ml）和万古霉素（4μg/ml）的MIC。此外，LHD蛋白可在不同pH值（6.0、7.0和8.0）下裂解……菌株。使用艰难梭菌感染（CDI）小鼠模型评估LHD的效力表明，给予LHD蛋白（每日两次，共7天）可有效减轻症状并降低CDI导致的死亡。用LHD治疗还显著降低了感染小鼠粪便中……芽孢的数量和毒素水平。我们的数据表明，这种新型溶菌酶-人防御素融合蛋白在控制CDI方面具有潜力。
