人工抗菌肽MA-D4的原核表达及其生物活性鉴定

Prokaryotic expression and biological activity identification of artifical antibacterial peptide MA-D4

为研究人工抗菌肽MA-D4的高效制备方法 ,检测其体外生物活性,探索该抗菌肽作为新型抗菌药物的应用前景和开发潜力,本试验设计人工抗菌肽MA-D4的氨基酸序列,并采用大肠杆菌偏嗜性密码子设计编码该抗菌肽的核苷酸序列;采取重叠区扩增基因拼接法合成目的基因,构建原核表达载体,并转化至大肠杆菌中诱导表达;表达产物经肠激酶酶切处理和镍离子亲和层析纯化后,进行体外生物活性检测。结果表明,人工抗菌肽MA-D4具备较强的广谱抗菌活性,可抑制革兰氏阳性菌和阴性菌,同时不具有溶血活性。综上所述,人工抗菌肽MA-D4在食品防腐、疾病治疗等方面具有良好的应用前景,极具开发潜力。

The test was conducted to study the efficient preparation methods of artificial antibacterial peptide MA-D4 and detect its biological activity in vitro, explore the application prospect and development potential of antibacterial peptide MA-D4 as a new type of antibacterial drug.The amino acid sequence of artificial antibacterial peptide MA-D4 was designed, and the nucleotide sequence coding antibacterial peptide MA-D4 was designed by using E.coli preferred codon; MA-D4 gene was spliced by overlap extension,then the prokaryotic expression vector was constructed and transformated; after the treatment of enterokinase and Ni ion affinity chromatography,the expression product was performanced the further detection of biological activity.The results showed that artificial antibacterial peptide MA-D4 had broad-spectrum antibacterial activity, could inhibit gram-positive bacterium and gram-negative bacteria, as well as had no hemolytic activity.The results indicated that artificial antibacterial peptide MA-D4 had good application prospect and development potential in such aspects as food preservative, disease treatment.