Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy.
Capsule-Targeting Depolymerase, Derived from Klebsiella KP36 Phage, as a Tool for the Development of Anti-Virulent Strategy.
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DOI：
10.3390/v8120324
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发表时间：
2016-12-01
期刊：
影响因子：
--
通讯作者：
Drulis-Kawa Z
中科院分区：
文献类型：
--
作者：
Majkowska-Skrobek G;Łątka A;Berisio R;Maciejewska B;Squeglia F;Romano M;Lavigne R;Struve C;Drulis-Kawa Z
抗生素耐药性肺炎克雷伯菌（一种主要的医院病原体）的增加促使人们需要替代疗法。我们鉴定并鉴定了一种由来自 Siphoviridae 家族的克雷伯氏菌噬菌体 KP36 (depoKP36) 编码的新型解聚酶。为了深入了解 depoKP36 的催化和结构特征，我们重组产生了这种 93.4 kDa 的蛋白质，并表明它能够水解肺炎克雷伯菌宿主的粗胞外多糖。通过体外和体内测定，我们发现 depoKP36 也能有效对抗临床肺炎克雷伯菌菌株（代表 K63 型）的天然荚膜，并以时间依赖性方式显着抑制克雷伯氏菌诱导的大蜡螟幼虫死亡率。 DepoKP36 不影响克雷伯氏菌菌株的抗生素敏感性。该酶的活性在较宽的 pH 值 (4.0–7.0) 和温度（最高 45 °C）范围内保持不变。一致的是，圆二色性 (CD) 光谱显示出高度稳定性，熔化转变温度 (Tm) = 65 °C。与其他噬菌体尾刺蛋白相比，该酶容易受到十二烷基硫酸钠 (SDS) 变性和蛋白水解切割的影响。溶液中的结构研究显示出具有高β-折叠含量的三聚体排列。我们的研究结果表明 depoKP36 是开发肺炎克雷伯菌感染新疗法的合适候选者。
The rise of antibiotic-resistant Klebsiella pneumoniae, a leading nosocomial pathogen, prompts the need for alternative therapies. We have identified and characterized a novel depolymerase enzyme encoded by Klebsiella phage KP36 (depoKP36), from the Siphoviridae family. To gain insights into the catalytic and structural features of depoKP36, we have recombinantly produced this protein of 93.4 kDa and showed that it is able to hydrolyze a crude exopolysaccharide of a K. pneumoniae host. Using in vitro and in vivo assays, we found that depoKP36 was also effective against a native capsule of clinical K. pneumoniae strains, representing the K63 type, and significantly inhibited Klebsiella-induced mortality of Galleria mellonella larvae in a time-dependent manner. DepoKP36 did not affect the antibiotic susceptibility of Klebsiella strains. The activity of this enzyme was retained in a broad range of pH values (4.0–7.0) and temperatures (up to 45 °C). Consistently, the circular dichroism (CD) spectroscopy revealed a highly stability with melting transition temperature (Tm) = 65 °C. In contrast to other phage tailspike proteins, this enzyme was susceptible to sodium dodecyl sulfate (SDS) denaturation and proteolytic cleavage. The structural studies in solution showed a trimeric arrangement with a high β-sheet content. Our findings identify depoKP36 as a suitable candidate for the development of new treatments for K. pneumoniae infections.
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影响因子：
4.8
作者：
Kęsik-Szeloch A;Drulis-Kawa Z;Weber-Dąbrowska B;Kassner J;Majkowska-Skrobek G;Augustyniak D;Lusiak-Szelachowska M;Zaczek M;Górski A;Kropinski AM
通讯作者：
Kropinski AM
影响因子：
4.2
作者：
Chhibber S;Nag D;Bansal S
通讯作者：
Bansal S
影响因子：
4.8
作者：
Hooton SP;Timms AR;Rowsell J;Wilson R;Connerton IF
通讯作者：
Connerton IF
影响因子：
3.6
作者：
Andres, Dorothee;Roske, Yvette;Barbirz, Stefanie
通讯作者：
Barbirz, Stefanie
影响因子：
3.1
作者：
DUTTON, GGS;DIFABIO, JL;STEPHEN, AM
通讯作者：
STEPHEN, AM