caa a22 4500 60549908X CHVBK 20210128100549.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6077-9 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6077-9 Design and recombination expression of a novel plectasin-derived peptide MP1106 and its properties against Staphylococcus aureus [Elektronische Daten] [Xintao Cao, Yong Zhang, Ruoyu Mao, Da Teng, Xiumin Wang, Jianhua Wang] A novel antimicrobial peptide MP1106 was designed based on the parental peptide plectasin with four mutational sites and a high level of expression in Pichia pastoris X-33 via the pPICZαA plasmid was achieved. The concentration of total secreted protein in the fermented supernatant was 2.134g/l (29°C), and the concentration of recombinant MP1106 (rMP1106) reached 1,808mg/l after a 120-h induction in a 5-l fermentor. The rMP1106 was purified using a cation-exchange column, and the yield was 831mg/l with 94.68% purity. The sample exhibited a narrow spectrum against some Gram-positive bacteria and strong antimicrobial activity against Staphylococcus aureus at low minimal inhibitory concentrations (MICs) of 0.014, 1.8, 0.45, and 0.91μM to S. aureus strains ATCC 25923, 29213, 6538, and 43300, respectively. Meanwhile, rMP1106 showed potent activity (0.03-1.8μM) against 20 clinical isolates of methicillin-resistant S. aureus (MRSA). In addition, rMP1106 exhibited a broad range of thermostability from 20 to 100°C. The higher antimicrobial activity of rMP1106 was maintained in neutral and alkaline environments (pH 6, 8, and 10), and its activity was slightly reduced in acidic environments (pH 2 and 4). The rMP1106 was resistant to the digestion of pepsin, snailase, and proteinase K and was sensitive to trypsin. It exhibited hemolytic activity of only 1.16% at a concentration of 512μg/ml and remained stable in human serum at 37°C for 24h. Furthermore, the activity of rMP1106 was minorly affected by 10mM dithiothreitol and 20% dimethylsulfoxide. Our results indicate that MP1106 can be produced on a large scale and has potential as a therapeutic drug against S. aureus. Springer-Verlag Berlin Heidelberg, 2014 MP1106 nationallicence Antimicrobial peptide nationallicence Pichia pastoris nationallicence Stability nationallicence Staphylococcus aureus nationallicence Cao Xintao Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Zhang Yong Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Mao Ruoyu Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Teng Da Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Wang Xiumin Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Wang Jianhua Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2649-2662 0175-7598 99:6<2649 2015 99 253 https://doi.org/10.1007/s00253-014-6077-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6077-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cao Xintao Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Zhang Yong Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Mao Ruoyu Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Teng Da Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Wang Xiumin Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Wang Jianhua Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2649-2662 0175-7598 99:6<2649 2015 99 253