The generation of nisin variants with enhanced activity against specific Gram-positive pathogens
The generation of nisin variants with enhanced activity against specific Gram-positive pathogens
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DOI：
10.1111/j.1365-2958.2008.06279.x
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发表时间：
2008-07-01
影响因子：
3.6
通讯作者：
Ross, R. Paul
中科院分区：
文献类型：
--
作者：
Field, Des;Connor, Paula M. O.;Ross, R. Paul
乳链菌肽是羊毛硫抗生素类抗菌肽的原型。它对革兰氏阳性菌表现出广谱抑制作用，包括重要的食品病原体和临床相关的耐药性细菌。值得注意的是，乳链菌肽的基因编码性质意味着它可以进行基于基因的生物工程以产生新的衍生物。在这里，我们利用这一点来生成迄今为止报道的最大的随机突变乳酸链球菌素衍生物库，最终目的是鉴定具有增强生物活性的变体。这种方法导致了乳酸链球菌素生产菌株的鉴定，该菌株具有增强的抗乳腺炎病原体无乳链球菌（Streptococcus agalactiae）的生物活性，这是由于肽（K22 T）的铰链区的氨基酸变化造成的。根据这一发现，采用铰链区残基的定点和位点饱和诱变，导致鉴定出另外的衍生物，最值得注意的是N20 P、M21 V和K22 S，其具有针对革兰氏阳性病原体（包括单核细胞增生李斯特菌和/或金黄色葡萄球菌）的增强的生物活性和比活性。这些衍生物的鉴定代表了乳链菌肽和羊毛硫抗生素的生物工程的一个重要步骤，并证实肽工程可以提供具有增强的抗微生物活性的衍生物，以对抗特定的有问题的腐败和病原微生物或对抗革兰氏阳性细菌。
Nisin is the prototype of the lantibiotic group of antimicrobial peptides. It exhibits broad spectrum inhibition of Gram-positive bacteria including important food pathogens and clinically relevant antibiotic-resistant bacteria. Significantly, the gene-encoded nature of nisin means that it can be subjected to gene-based bioengineering to generate novel derivatives. Here, we take advantage of this to generate the largest bank of randomly mutated nisin derivatives reported to date, with the ultimate aim of identifying variants with enhanced bioactivity. This approach led to the identification of a nisin-producing strain with enhanced bioactivity against the mastitic pathogen Streptococcus agalactiae resulting from an amino acid change in the hinge region of the peptide (K22T). Prompted by this discovery, site-directed and site-saturation mutagenesis of the hinge region residues was employed, resulting in the identification of additional derivatives, most notably N20P, M21V and K22S, with enhanced bioactivity and specific activity against Gram-positive pathogens including Listeria monocytogenes and/or Staphylococcus aureus. The identification of these derivatives represents a major step forward in the bioengineering of nisin, and lantibiotics in general, and confirms that peptide engineering can deliver derivatives with enhanced antimicrobial activity against specific problematic spoilage and pathogenic microbes or against Gram-positive bacteria in general.