Rao Zhili, Kim So Young, Akanda Md Rashedunnabi, Lee Su Jin, Jung In Duk, Park Byung-Yong, Kamala-Kannan Seralathan, Hur Jin, Park Jung Hee
Division of Biotechnology, College of Environmental & Bioresources Sciences, Chonbuk National University, Iksan 54596, Korea.
College of Veterinary Medicine and Bio-safety Research Institute, Chonbuk National University, Iksan 54596, Korea.
Mol Cells. 2019 Mar 31;42(3):262-269. doi: 10.14348/molcells.2019.2365. Epub 2019 Feb 19.
The porcine myeloid antimicrobial peptide (PMAP), one of the cathelicidin family members, contains small cationic peptides with amphipathic properties. We used a putative lysozyme originated from the bacteriophage P22 (P22 lysozyme) as a fusion partner, which was connected to the N-terminus of the PMAP36 peptide, to markedly increase the expression levels of recombinant PMAP36. The PMAP36-P22 lysozyme fusion protein with high solubility was produced in . The final purified yield was approximately 1.8 mg/L. The purified PMAP36-P22 lysozyme fusion protein exhibited antimicrobial activity against both Gram-negative and Gram-positive bacteria (, serovar Typhimurium, , and ). Furthermore, we estimated its hemolytic activity against pig erythrocytes as 6% at the high concentration (128 μM) of the PMAP36-P22 lysozyme fusion protein. Compared with the PMAP36 peptide (12%), our fusion protein exhibited half of the hemolytic activity. Overall, our recombinant PMAP36-P22 lysozyme fusion protein sustained the antimicrobial activity with the lower hemolytic activity associated with the synthetic PMAP36 peptide. This study suggests that the PMAP36-P22 lysozyme fusion system could be a crucial addition to the plethora of novel antimicrobials.
猪髓样抗菌肽（PMAP）是组织蛋白酶抗菌肽家族成员之一，由具有两亲性的小阳离子肽组成。我们使用一种源自噬菌体P22的假定溶菌酶（P22溶菌酶）作为融合伴侣，将其连接到PMAP36肽的N端，以显著提高重组PMAP36的表达水平。在……中产生了具有高溶解度的PMAP36-P22溶菌酶融合蛋白。最终纯化产量约为1.8 mg/L。纯化后的PMAP36-P22溶菌酶融合蛋白对革兰氏阴性菌和革兰氏阳性菌（如鼠伤寒沙门氏菌血清型、……和……）均表现出抗菌活性。此外，我们估计在高浓度（128 μM）的PMAP36-P22溶菌酶融合蛋白作用下，其对猪红细胞的溶血活性为6%。与PMAP36肽（12%）相比，我们的融合蛋白溶血活性降低了一半。总体而言，我们的重组PMAP36-P22溶菌酶融合蛋白在保持抗菌活性的同时，与合成的PMAP36肽相比具有较低的溶血活性。这项研究表明，PMAP36-P22溶菌酶融合系统可能是众多新型抗菌剂中的一个重要补充。
