东北黑斑蛙皮肤抗菌肽的分离及抗菌活性检测

Isolation and antibacterial activity detection of antibacterial peptides from skin of Rana nigromaculata

从成年及幼年两组东北黑斑蛙皮肤分泌物中分离抗菌肽,并对其抗菌活性进行检测。采用电刺激法,收集黑斑蛙皮肤分泌物,应用反相高效液相色谱仪进行分离,应用MALDI-TOF/TOF质谱仪鉴定,通过最小抑菌浓度(MIC)试验,检测抗菌肽的抑菌效果。结果表明,AMP1及AMP2的抑菌作用较好,相对分子量分别为3 349.205 Da及1 373.820 Da。AMP1的MIC值分别为耐甲氧西林金黄色葡萄球菌(MRSA)16μg/ml、金黄色葡萄球菌8μg/ml、枯草芽孢杆菌64μg/ml、肠炎耶尔森氏菌16μg/ml、肺炎克雷伯菌64μg/ml、大肠杆菌8μg/ml;AMP2的MIC值分别为金黄色葡萄球菌32μg/ml、枯草芽孢杆菌128μg/ml、肠炎耶尔森氏菌32μg/ml、大肠杆菌16μg/ml。试验结果表明,分离得到的抗菌肽具有广谱抗菌性,具有一定的开发潜力。

The skin antibacterial peptides were extracted from senior group and infant group of wildnortheast Rana nigromaculata, and the antibacterial activity was deteted. The experiment was done bythe electrical stimulation, collectting skin secretions, isolataing and identifing by RP-HPLC and MAL-DI-TOF/TOF. Through the further minimal inhibitory concentration（MIC） to test their antibacterial ac-tivity. There were two peptides AMP1 and AMP2 possessing powerful antibacterial activity. The molec-ular weight were 3 349.205 Da and 1 373.820 Da. The MIC rate of AMP1 was MRSA 16 μg/ml, S.au-reus ATC29213 8 μg/ml, Bacillus subtilis 64 μg/ml, Yersinia enterocolitica 16 μg/ml, Klebsiella pneu-moniae 64 μg/ml, E.coli ATC25922 8 μg/ml. AMP2 was S.aureus ATC29213 32 μg/ml, Bacillus subti-lis 128 μg/ml, Yersinia enterocolitica 32 μg/ml, E.coli ATC25922 16 μg/ml. The result revealved thatthe antibacterial peptides showed the broad-spectrum antibacterial activity and a certain potential.