Discovery of novel S. aureus autolysins and molecular engineering to enhance bacteriolytic activity.
Discovery of novel S. aureus autolysins and molecular engineering to enhance bacteriolytic activity.
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DOI：
10.1007/s00253-015-6443-2
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发表时间：
2015-08
影响因子：
5
通讯作者：
Griswold, Karl E.
中科院分区：
文献类型：
--
作者：
Osipovitch, Daniel C.;Therrien, Sophie;Griswold, Karl E.
金黄色葡萄球菌是一种危险的细菌病原体，其临床影响因抗生素耐药性的出现和迅速传播而放大。在寻找更有效的治疗策略的过程中，人们在葡萄球菌分解酶的研究和开发上投入了大量精力，这种酶具有高效力、对耐药菌株的活性以及对新耐药表型出现的低固有敏感性。迄今为止，大多数候选治疗药物都源自金黄色葡萄球菌的噬菌体或环境竞争者。几乎没有考虑到顺式作用自溶素，它代表了细菌内源细胞壁维护和回收机制的关键要素。在这项研究中，从金黄色葡萄球菌基因组中克隆了五种假定的自溶素，并评估了它们的活性。这些新型酶中的四种或其组成域表现出对活金黄色葡萄球菌细胞的裂解活性，但它们的效力比充分表征的治疗候选溶葡萄球菌素低数十至数千倍。我们假设它们的活性较差部分是由于与其天然细胞壁结合域相关的细胞壁靶向性欠佳，并且我们试图通过与溶葡萄球菌素的肽聚糖结合域嵌合来增强它们的抗菌潜力。最有效的嵌合体的溶解速率增加了 140 倍，与溶葡萄球菌素的溶解速率相差不到 8 倍。虽然这种酶对某些生物学相关的环境因素敏感并且未能表现出可测量的最小抑制浓度，但它能够杀死耐溶葡萄球菌素的金黄色葡萄球菌，并最终证明在肺表面活性剂中具有活性。我们得出的结论是，金黄色葡萄球菌蛋白质组代表了丰富且未开发的新型抗菌酶库，并且我们证明通过改进自溶素催化结构域的细胞壁靶向来增强溶菌活性。
Staphylococcus aureus is a dangerous bacterial pathogen whose clinical impact has been amplified by the emergence and rapid spread of antibiotic resistance. In the search for more effective therapeutic strategies, great effort has been placed on the study and development of staphylolytic enzymes, which benefit from high potency, activity towards drug-resistant strains, and a low inherent susceptibility to emergence of new resistance phenotypes. To date, the majority of therapeutic candidates have derived from either bacteriophage or environmental competitors of S. aureus. Little to no consideration has been given to cis-acting autolysins that represent key elements in the bacterium's endogenous cell wall maintenance and recycling machinery. In this study, five putative autolysins were cloned from the S. aureus genome, and their activities were evaluated. Four of these novel enzymes, or component domains thereof, demonstrated lytic activity towards live S. aureus cells, but their potencies were 10s to 1000s of times lower than that of the well-characterized therapeutic candidate lysostaphin. We hypothesized that their poor activities were due in part to suboptimal cell wall targeting associated with their native cell wall binding domains, and we sought to enhance their antibacterial potential via chimeragenesis with the peptidoglycan binding domain of lysostaphin. The most potent chimera exhibited a 140-fold increase in lytic rate, bringing it within 8-fold of lysostaphin. While this enzyme was sensitive to certain biologically relevant environmental factors and failed to exhibit a measurable minimal inhibitory concentration, it was able to kill lysostaphin-resistant S. aureus and ultimately proved active in lung surfactant. We conclude that the S. aureus proteome represents a rich and untapped reservoir of novel antibacterial enzymes, and we demonstrate enhanced bacteriolytic activity via improved cell wall targeting of autolysin catalytic domains.
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