Design and pharmacodynamics of recombinant NZ2114 histidine mutants with improved activity against methicillin-resistant Staphylococcus aureus.
Design and pharmacodynamics of recombinant NZ2114 histidine mutants with improved activity against methicillin-resistant Staphylococcus aureus.
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重组NZ2114组氨酸突变体的设计和药效学，其对耐甲氧西林金黄色葡萄球菌具有改善的活性
DOI：
10.1186/s13568-017-0345-x
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发表时间：
2017-12
期刊：
影响因子：
3.7
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Chen H;Mao R;Teng D;Wang X;Hao Y;Feng X;Wang J
NZ 2114对金黄色葡萄球菌具有较强的抗菌活性，是一种很有前途的药物。我们的目的是通过用精氨酸和赖氨酸残基取代His 16和His 18来鉴定具有改进活性的NZ 2114衍生物。设计了8个突变体，并通过pPICZαA在毕赤酵母X-33中表达。其中5个化合物对沙门氏菌有较强的抗菌活性。金黄色葡萄球菌在0.057-0.454 μM的低最小抑菌浓度（MIC）下。其中H1、H2和H3对耐甲氧西林沙门氏菌有较好的药效作用。aureus ATCC 43300。在5 L发酵罐中诱导120 h时，H1、H2和H3的总蛋白水平分别达到1.70、1.77和1.54 g/l。在2×和4× MIC时，1.5 h内杀灭率达99.9%以上。H1、H2和H3对S. aureus ATCC 43300在2× MIC时的作用时间分别为2.94、1.75和1.55 h，与其原始肽NZ 2114（1.43 h）和万古霉素（1.72 h）相似。H1、H2、H3与万古霉素、氨苄青霉素、利福平的抑菌浓度指数（FICI）无显著性差异。此外，它们在不同环境（温度、pH、蛋白酶和盐离子）中具有低溶血性和高稳定性。所有结果表明，H1、H2和H3可以大规模生产，并具有作为抗MRSA治疗药物的潜力。
NZ2114 is a promising candidate for therapeutic application owing to its potent activity to Staphylococcus aureus. Our objective was to identify NZ2114 derivatives with improved activity through substitution of His16 and His18 with residues Arginine and Lysine. Eight mutants were designed and expressed in Pichia pastoris X-33 via pPICZαA. Five of them exhibited strong antimicrobial activity against S. aureus at low minimal inhibitory concentrations (MICs) of 0.057-0.454 μM. Among them, H1, H2, and H3 showed ideal pharmacodynamic effects on methicillin-resistant S. aureus ATCC43300. The total protein level of H1, H2, and H3 reached 1.70, 1.77 and 1.54 g/l at 120 h of induction in the 5-l fermenter, respectively. They killed over 99.9% of pathogens within 1.5 h at 2× and 4× MIC. The post antibiotic effect of H1, H2 and H3 to S. aureus ATCC43300 was 2.94, 1.75 and 1.55 h at 2× MIC, which was similar with their original peptide NZ2114 (1.43 h) and vancomycin (1.72 h). The fractional inhibitory concentration index (FICI) indicated indifferent effects between H1, H2, H3 and vancomycin, ampicillin, rifampicin. Additionally, they had low hemolysis and high stability in different environments (temperature, pH, proteases, and saline ions). All results indicate that H1, H2, and H3 can be produced in large-scale and have potential as therapeutic drugs against MRSA.