IMPROVEMENT OF SOLUBILITY AND STABILITY OF THE ANTIMICROBIAL PEPTIDE NISIN BY PROTEIN ENGINEERING
IMPROVEMENT OF SOLUBILITY AND STABILITY OF THE ANTIMICROBIAL PEPTIDE NISIN BY PROTEIN ENGINEERING
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DOI：
10.1128/aem.61.8.2873-2878.1995
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发表时间：
1995-08-01
影响因子：
4.4
通讯作者：
SIEZEN, RJ
中科院分区：
文献类型：
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作者：
ROLLEMA, HS;KUIPERS, OP;SIEZEN, RJ
Nisin 是一种 3.4 kDa 的抗菌肽，经过翻译后修饰，含有不饱和氨基酸和羊毛硫氨酸残基，可用作各种食品中的防腐剂。已经研究了乳链菌肽和乳链菌肽突变体的溶解度和稳定性。事实证明，可以产生具有改进的功能特性的乳链菌肽突变体。乳链菌肽 A 的溶解度在低 pH 值时最高，当溶液的 pH 值超过 7 时，溶解度逐渐降低近 2 个数量级。在低 pH 值下，乳链菌肽 Z 的溶解度相对于乳链菌肽 A 表现出降低；在中性和较高 pH 值下，两种变体的溶解度相当。生产了两种乳链菌肽 Z 突变体，分别在位置 27 和 31 处含有赖氨酰残基，而不是 Asn-27 和 His-31，其目的是在中性 pH 下达到更高的溶解度。两种突变体均纯化至均质，并通过一维和二维 H-1 核磁共振证实了其结构，发现其抗菌活性与乳链菌肽 Z 相似，而它们在 pH 7 下的溶解度分别增加了 4 倍和 7 倍。 研究了乳链菌肽 A 在 pH 2 至 8 范围内以及 20、37 和 75 ℃ 下的化学稳定性，在 pH 下观察到最佳稳定性在 3.0 版本中，乳链菌肽 Z 显示出与乳链菌肽 A 类似的行为。在第 5 位含有脱氢丁酸而不是脱氢丙氨酸的突变体具有较低的活性，但比野生型乳链菌肽 Z 更能抵抗酸催化的化学降解。
Nisin is a 3.4-kDa antimicrobial peptide that, as a result of posttranslational modifications, contains unsaturated amino acids and lanthionine residues, It is applied as a preservative in various food products. The solubility and stability of nisin and nisin mutants have been studied. It is demonstrated that nisin mutants can be produced with improved functional properties. The solubility of nisin A is highest at low pH values and gradually decreases by almost 2 orders of magnitude when the pH of the solution exceeds a value of 7, At low pH, nisin Z exhibits a decreased solubility relative to that of nisin A; at neutral and higher pH values, the solubilities of both variants are comparable. Two mutants of nisin Z, which contain lysyl residues at positions 27 and 31, respectively, instead of Asn-27 and His-31, were produced with the aim of reaching higher solubility at neutral pH. Both mutants were purified to homogeneity, and their structures were confirmed by one- and two-dimensional H-1 nuclear magnetic resonance, Their antimicrobial activities were found to be similar to that of nisin Z, whereas their solubilities at pH 7 increased by factors of 4 and 7, respectively, The chemical stability of nisin A was studied in the pH range of 2 to 8 and at 20, 37, and 75 degrees C, Optimal stability was observed at pH 3.0, Nisin Z showed a behavior similar to that of nisin A. A mutant containing dehydrobutyrine at position 5 instead of dehydroalanine had lower activity but was significantly more resistant to acid-catalyzed chemical degradation than wild-type nisin Z.