Lytic enzyme discovery through multigenomic sequence analysis in Clostridium perfringens.
Lytic enzyme discovery through multigenomic sequence analysis in Clostridium perfringens.
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DOI：
10.1007/s00253-010-2982-8
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发表时间：
2011-03
影响因子：
5
通讯作者：
Fischetti, Vincent A.
中科院分区：
文献类型：
--
作者：
Schmitz, Jonathan E.;Ossiprandi, Maria Cristina;Rumah, Kareem R.;Fischetti, Vincent A.
由于具有裂解革兰氏阳性菌的能力，噬菌体裂解酶（或溶素）作为新型抗感染剂受到了广泛关注。近年来，编码这些肽聚糖水解酶的已知基因数量显着增加，这在很大程度上归功于 DNA 测序技术的进步。随着越来越多的细菌物种/菌株的基因组被测序，可以在溶原化的原噬菌体区域中轻松地识别出编码溶素的开放阅读框（ORF）。在当前的研究中，我们试图评估医学相关病原体产气荚膜梭菌的溶素多样性。通过计算挖掘九种产气荚膜梭菌菌株的基因组序列，找出原噬菌体溶素和类溶素 ORF，揭示了数十种不同酶类的蛋白质。在这些溶素中，选择来自菌株 ATCC 13124 的胞壁质酶（称为 PlyCM）进行重组分析，因为它与先前表征的产气荚膜梭菌溶素不同。表达和纯化后，在体外测定了 PlyCM 的各种生化特性，包括 pH/盐依赖性和温度稳定性。该酶在低 µg/ml 浓度（噬菌体溶素的典型值）下表现出活性。它对所测试的 24 种产气荚膜梭菌菌株中的 23 种有活性，而对其他梭菌或非梭菌物种几乎没有活性。总体而言，PlyCM 显示出作为酶生剂的开发潜力，展示了不断扩大的基因组数据库如何成为生物技术相关蛋白质的丰富库。
With their ability to lyse Gram-positive bacteria, phage lytic enzymes (or lysins) have received a great deal of attention as novel anti-infective agents. The number of known genes encoding these peptidoglycan hydrolases has increased markedly in recent years, due in large part to advances in DNA sequencing technology. As the genomes of more and more bacterial species/strains are sequenced, lysin-encoding open reading frames (ORFs) can be readily identified in lysogenized prophage regions. In the current study, we sought to assess lysin diversity for the medically relevant pathogen Clostridium perfringens. The sequenced genomes of nine C. perfringens strains were computationally mined for prophage lysins and lysin-like ORFs, revealing several dozen proteins of various enzymatic classes. Of these lysins, a muramidase from strain ATCC 13124 (termed PlyCM) was chosen for recombinant analysis based on its dissimilarity to previously characterized C. perfringens lysins. Following expression and purification, various biochemical properties of PlyCM were determined in vitro, including pH/salt-dependence and temperature stability. The enzyme exhibited activity at low µg/ml concentrations, a typical value for phage lysins. It was active against 23 of 24 strains of C. perfringens tested, with virtually no activity against other clostridial or nonclostridial species. Overall, PlyCM shows potential for development as an enzybiotic agent, demonstrating how expanding genomic databases can serve as rich pools for biotechnologically relevant proteins.
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影响因子：
4.1
作者：
Fischetti, Vincent A.
通讯作者：
Fischetti, Vincent A.
影响因子：
4.4
作者：
Deutsch, SM;Guezenec, S;Lortal, S
通讯作者：
Lortal, S
影响因子：
14.9
作者：
Finn RD;Mistry J;Tate J;Coggill P;Heger A;Pollington JE;Gavin OL;Gunasekaran P;Ceric G;Forslund K;Holm L;Sonnhammer EL;Eddy SR;Bateman A
通讯作者：
Bateman A
影响因子：
5.7
作者：
Hermoso, JA;Monterroso, B;Menéndez, M
通讯作者：
Menéndez, M
DOI：
10.1128/aem.28.4.655-660.1974
发表时间：
1974-01-01
期刊：
APPLIED MICROBIOLOGY
影响因子：
--
作者：
MATCHES, JR;LISTON, J;CURRAN, D
通讯作者：
CURRAN, D