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Appl Microbiol Biotechnol

Title:		"Design, expression, and characterization of a novel targeted plectasin against methicillin-resistant Staphylococcus aureus"
Author(s):		Mao R; Teng D; Wang X; Xi D; Zhang Y; Hu X; Yang Y; Wang J;
Address:		"Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Beijing 100081, China"
Journal Title:		Appl Microbiol Biotechnol
Year:		2013
Volume:		20121025
Issue:		9
Page Number:		3991 - 4002
DOI:		10.1007/s00253-012-4508-z
ISSN/ISBN:		1432-0614 (Electronic) 0175-7598 (Linking)
Abstract:		"A novel specifically targeted antimicrobial peptide (STAMP) that was especially effective against methicillin-resistant Staphylococcus aureus (MRSA) was designed by fusing the AgrD1 pheromone to the N-terminal end of plectasin. This STAMP was named Agplectasin, and its gene was synthesized and expressed in Pichia pastoris X-33 via pPICZalphaA. The highest amount of total secreted protein reached 1,285.5 mg/l at 108 h during the 120-h induction. The recombinant Agplectasin (rAgP) was purified by cation exchange chromatography and hydrophobic exchange chromatography; its yield reached 150 mg/l with 94 % purity. The rAgP exhibited strong bactericidal activity against S. aureus but not Staphylococcus epidermidis or other types of tested bacteria. A bactericidal kinetics assay showed that the rAgP killed over 99.9 % of tested S. aureus (ATCC 25923 and ATCC 43300) in both Mueller-Hinton medium and human blood within 10 h when treated with 4x minimal inhibitory concentration. The rAgP caused only approximately 1 % hemolysis of human blood cells, even when the concentration reached 512 mug/ml, making it potentially feasible as a clinical injection agent. In addition, it maintained a high activity over a wide range of pH values (2.0-10.0) and demonstrated a high thermal stability at 100 degrees C for 1 h. These results suggested that this STAMP has the potential to eliminate MRSA strains without disrupting the normal flora"
Keywords:		"Anti-Bacterial Agents/*metabolism/*pharmacology Bacterial Proteins/genetics/*metabolism Methicillin-Resistant Staphylococcus aureus/*drug effects Microbial Sensitivity Tests Peptides/genetics/*metabolism Peptides, Cyclic/genetics/*metabolism Pichia/geneti;"
Notes:		"MedlineMao, Ruoyu Teng, Da Wang, Xiumin Xi, Di Zhang, Yong Hu, Xiaoyuan Yang, Yalin Wang, Jianhua eng Research Support, Non-U.S. Gov't Germany 2012/10/26 Appl Microbiol Biotechnol. 2013 May; 97(9):3991-4002. doi: 10.1007/s00253-012-4508-z. Epub 2012 Oct 25"