Niu Ming-Fu, Li Xiang, Cao Rui-Bing, Zhou Bin, Chen De-Sheng, Chen Pu-Yan
Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
Sheng Wu Gong Cheng Xue Bao. 2007 May;23(3):418-22.
In order to obtain a high activity antibacterial peptide, An expression vector pPICZalphaA-pl is constructed with a tandem of four antimicrobial peptides in the same direction,which includes Protegrin-1 (PG-1), Scorpion Defensin (SD), Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide (SMAP-29) (serial number in GenBank are AAB27599, AAAB27538, P80409 and P49928 respectively). At the same time the expression vector pPICZalphaA-sd which express Scorpion Defensin was contructed. The expression vectors of pPICZalphaA-pl and pPICZalphaA-sd were linearized and transformed into the yeast host strain X-33 respectively. Under the control of the promoter AOX1 (alcohol oxidase1), the peptides PL and SD were secreted expressed. Their heat-stable property, acid-stable property and MIC were detected in vitro. The results suggest the peptides PL and SD have good heat-stable and acid-stable properties, and the combinant PL peptide showes higher antibacterial activity against several Gram-positive bacteria (G+) and Gram-negative bacteria (G-) than the peptide SD, especially against Escherichia coli. The antibacterial activity of combinant antimicrobial peptide PL shows its far exploiting perspective.
为了获得高活性抗菌肽,构建了一种表达载体pPICZalphaA-pl,其串联了四个同向的抗菌肽,包括Protegrin-1(PG-1)、蝎防御素(SD)、Metalnikowin-2A和绵羊髓系抗菌肽(SMAP-29)(在GenBank中的序列号分别为AAB27599、AAAB27538、P80409和P49928)。同时构建了表达蝎防御素的表达载体pPICZalphaA-sd。将pPICZalphaA-pl和pPICZalphaA-sd表达载体分别线性化并转化到酵母宿主菌株X-33中。在启动子AOX1(乙醇氧化酶1)的控制下,肽PL和SD被分泌表达。体外检测了它们的热稳定性、酸稳定性和最小抑菌浓度(MIC)。结果表明,肽PL和SD具有良好的热稳定性和酸稳定性,组合肽PL对几种革兰氏阳性菌(G+)和革兰氏阴性菌(G-)的抗菌活性高于肽SD,尤其是对大肠杆菌。组合抗菌肽PL的抗菌活性显示出其广阔的开发前景。