Intensive mutagenesis of the nisin hinge leads to the rational design of enhanced derivatives.
Intensive mutagenesis of the nisin hinge leads to the rational design of enhanced derivatives.
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DOI：
10.1371/journal.pone.0079563
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ross RP
中科院分区：
文献类型：
--
作者：
Healy B;Field D;O'Connor PM;Hill C;Cotter PD;Ross RP
乳链菌肽 A 是研究最广泛的羊毛硫抗生素，自 1953 年以来一直被食品工业用作防腐剂。这种 34 个氨基酸的肽包含三个脱水氨基酸和五个硫醚环。这些环由一个羊毛硫氨酸和四个甲基羊毛硫氨酸桥组成，赋予肽独特的结构。乳链菌肽 A 有两种作用机制，肽的 N 端结构域通过脂质 II 结合抑制细胞壁合成，C 端结构域负责孔的形成。这项研究的重点是三个氨基酸“铰链”区域（N 20、M 21 和 K 22），它将这两个结构域分开并允许构象灵活性。由于所有羊毛硫抗生素都是基因编码的，因此可以通过操作相应的基因来产生新的变体。先前已证明许多铰链区被改变的衍生物具有增强的抗菌活性。在这里，我们进一步采用这种方法，对所有三个铰链残基采用同时、不加区别的位点饱和诱变，以创建新型乳链菌肽衍生物生产者库。对该库的筛选表明，具有由 AAK、NAI 和 SLS 组成的铰链区的肽的生产商对多种靶点表现出增强的生物活性。这些和其他结果表明铰链区内对小手性氨基酸的偏好，导致设计和创建具有由 AAA 和 SAA 组成的铰链的肽生产商。这些生产者和相应的肽对乳酸乳球菌 HP、无乳链球菌 ATCC 13813、耻垢分枝杆菌 MC2155 和金黄色葡萄球菌 RF122 表现出增强的生物活性，因此代表了通过合理设计而具有增强活性的乳链菌肽衍生物的第一个例子。
Nisin A is the most extensively studied lantibiotic and has been used as a preservative by the food industry since 1953. This 34 amino acid peptide contains three dehydrated amino acids and five thioether rings. These rings, resulting from one lanthionine and four methyllanthionine bridges, confer the peptide with its unique structure. Nisin A has two mechanisms of action, with the N-terminal domain of the peptide inhibiting cell wall synthesis through lipid II binding and the C-terminal domain responsible for pore-formation. The focus of this study is the three amino acid ‘hinge’ region (N 20, M 21 and K 22) which separates these two domains and allows for conformational flexibility. As all lantibiotics are gene encoded, novel variants can be generated through manipulation of the corresponding gene. A number of derivatives in which the hinge region was altered have previously been shown to possess enhanced antimicrobial activity. Here we take this approach further by employing simultaneous, indiscriminate site-saturation mutagenesis of all three hinge residues to create a novel bank of nisin derivative producers. Screening of this bank revealed that producers of peptides with hinge regions consisting of AAK, NAI and SLS displayed enhanced bioactivity against a variety of targets. These and other results suggested a preference for small, chiral amino acids within the hinge region, leading to the design and creation of producers of peptides with hinges consisting of AAA and SAA. These producers, and the corresponding peptides, exhibited enhanced bioactivity against Lactococcus lactis HP, Streptococcus agalactiae ATCC 13813, Mycobacterium smegmatis MC2155 and Staphylococcus aureus RF122 and thus represent the first example of nisin derivatives that possess enhanced activity as a consequence of rational design.
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影响因子：
3.6
作者：
Brötz, H;Josten, M;Sahl, HG
通讯作者：
Sahl, HG
影响因子：
4.8
作者：
Lubelski, Jacek;Khusainov, Rustem;Kuipers, Oscar P.
通讯作者：
Kuipers, Oscar P.
影响因子：
4.4
作者：
de Kwaadsteniet, M.;ten Doeschate, K.;Dicks, L. M. T.
通讯作者：
Dicks, L. M. T.
影响因子：
3.7
作者：
Field D;Begley M;O'Connor PM;Daly KM;Hugenholtz F;Cotter PD;Hill C;Ross RP
通讯作者：
Ross RP
影响因子：
16.8
作者：
Hsu, STD;Breukink, E;van Nuland, NAJ
通讯作者：
van Nuland, NAJ