caa a22 4500 605506604 CHVBK 20210128100628.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6394-7 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6394-7 In vitro and invivo characterization of a new recombinant antimicrobial peptide, MP1102, against methicillin-resistant Staphylococcus aureus [Elektronische Daten] [Yong Zhang, Da Teng, Xiumin Wang, Ruoyu Mao, Xintao Cao, Xiaoyuan Hu, Lifen Zong, Jianhua Wang] Currently, more antimicrobial drug candidates are urgently needed to combat the rise in drug-resistance among pathogenic microbes. A new antimicrobial peptide, MP1102, a variant of NZ2114, was designed, evaluated, and overexpressed in Pichia pastoris. The total secreted protein in cultures reached 695mg/l, and the concentration of the recombinant MP1102 (rMP1102) was 292mg/l. rMP1102 was purified from the fermentation supernatant by one-step cation exchange chromatography to obtain a yield of 197.1mg/l with 96.4% purity. rMP1102 exhibited potent activity against Gram-positive bacteria, and its minimum inhibitory concentrations (MICs) for four Staphyloccocus aureus (S. aureus) strains ranged from 0.028 to 0.11μM, and it had stronger activity (MIC = 0.04 to 0.23μM) to 20 clinical isolates of MRSA (cMRSA) than rNZ2114 (MIC = 0.11 to 0.90μM). rMP1102 was shown to kill over 99.9% of tested S. aureus cells within 6h when treated at one, two, and four times its MIC and over 90% of S. aureus cells within 12h at concentrations of 5, 10, and 20mg/kg in a mouse thigh infection model. The higher sensitivity of MRSA to MP1102 than to its parental peptide, NZ2114, indicated by this initial pharmacodynamic analysis suggests a possible difference in the killing mechanism of these two molecules. rMP1102 caused less than 0.05% hemolytic activity at 128μg/ml and exhibited good thermostability from 20 to 80°C, with its highest activity being observed at pH8.0. These results suggest that this yeast expression system is feasible for large-scale production, and rMP1102 exerted stronger activity against S. aureus than NZ2114 via a different mechanism and exhibited potential as a new antimicrobial agent for S. aureus, especially MRSA infections. Springer-Verlag Berlin Heidelberg, 2015 Molecular design nationallicence MP1102 nationallicence Pichia pastoris nationallicence MRSA nationallicence Antimicrobial peptides nationallicence Zhang Yong 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 Mao Ruoyu Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Cao Xintao Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Hu Xiaoyuan Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut Zong Lifen 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 99/15(2015-08-01), 6255-6266 0175-7598 99:15<6255 2015 99 253 https://doi.org/10.1007/s00253-015-6394-7 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/s00253-015-6394-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Yong 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- Mao Ruoyu Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Cao Xintao Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Hu Xiaoyuan Key Laboratory of Feed Biotechnology, Ministry of Agriculture, 100081, Beijing, China aut NATIONALLICENCE 700 1- Zong Lifen 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 99/15(2015-08-01), 6255-6266 0175-7598 99:15<6255 2015 99 253