title: Design and recombination expression of a novel plectasin-derived peptide MP1106 and its properties against Staphylococcus aureus.
abstract: 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 2.134 g/l (29 °C), and the concentration of recombinant MP1106 (rMP1106) reached 1,808 mg/l after a 120-h induction in a 5-l fermentor. The rMP1106 was purified using a cation-exchange column, and the yield was 831 mg/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 0.91 μM to S. aureus strains ATCC 25923, 29213, 6538, and 43300, respectively. Meanwhile, rMP1106 showed potent activity (0.03-1.8 μM) 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 512 μg/ml and remained stable in human serum at 37 °C for 24 h. Furthermore, the activity of rMP1106 was minorly affected by 10 mM 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.
authors.affiliation: Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Beijing, 100081, China.
authors.forename: Xintao
authors.initials: X
authors.lastname: Cao
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Mao
authors.forename: Da
authors.initials: D
authors.lastname: Teng
authors.forename: Xiumin
authors.initials: X
authors.lastname: Wang
authors.forename: Jianhua
authors.initials: J
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001483:Base Sequence; D004912:Erythrocytes / Q000187:drug effects; D006094:Gram-Positive Bacteria; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000494:pharmacology*; D010843:Pichia / Q000378:metabolism; D011487:Protein Conformation; D015202:Protein Engineering
chemical_list: D000900:Anti-Bacterial Agents; D010455:Peptides; C504857:plectasin