Zhou Qingfeng, Li Mingyue, Xi Tao
Department of Life Science and Technology, Shangqiu Normal University, Shangqiu, China.
Curr Microbiol. 2009 Apr;58(4):384-8. doi: 10.1007/s00284-009-9372-4. Epub 2009 Feb 7.
Perinerin, an antimicrobial peptide (GenBank No. P84117), was isolated and characterized from Asian marine clamworms, Perinereis aibuhitensis Grube. This peptide has effects against both gram-positive and gram-negative bacteria in vitro, especially on Pseudemonas aeruginosa. In our study, bioactive perinerin was expressed in Pichia pastoris, and characterized its physicochemical properties. The expressed sample was firstly analyzed by Tricine-SDS-PAGE, after then the recombinant proteins were purified by 2 kD MW cut-off (MWCO) ultrafiltration membrane, and finally purified by 10 kD MWCO ultrafiltration membrane and CM 52-ion exchange chromatography. About 6% protein was obtained by this so called three-purification method. Our results showed that Pichia pastoris was a suitable system for secreting perinerin. Bioactivity assay proved that the recombinant perinerin had antimicrobial effects.
围沙蚕抗菌肽（GenBank登录号：P84117）是从亚洲海蚯蚓，即双齿围沙蚕中分离并鉴定出来的。该抗菌肽在体外对革兰氏阳性菌和革兰氏阴性菌均有作用，尤其对铜绿假单胞菌效果显著。在本研究中，具有生物活性的围沙蚕抗菌肽在毕赤酵母中表达，并对其理化性质进行了表征。表达产物首先通过Tricine-SDS-PAGE进行分析，然后用截留分子量为2 kD的超滤膜对重组蛋白进行纯化，最后用截留分子量为10 kD的超滤膜和CM 52离子交换层析进一步纯化。通过这种所谓的三步纯化法获得了约6%的蛋白。我们的结果表明，毕赤酵母是分泌围沙蚕抗菌肽的合适系统。生物活性测定证明重组围沙蚕抗菌肽具有抗菌作用。
