The molecular design of a recombinant antimicrobial peptide CP and its in vitro activity.
Antibacterial peptides from various sources express different antibacterial activity. In order to obtain a high activity antibacterial peptide, the sequences of four antimicrobial peptides--Protegrin-1, 4 kDa Scorpion Defensin, Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide SMAP-29--were exploited to generate a synthetic antimicrobial peptide cp gene, which was then cloned into the expression vector pPICZalpha-A. The constructed recombinant expression vector pPICZalpha-cp was transformed into Pichia pastoris X-33, in which the synthetic antimicrobial peptide (CP) could be expressed under the control of the inducible AOX1 promoter and secreted via the alpha mating factor leader of Saccharomyces cerevisiae. Results showed that recombinant plasmid is highly stable, and In vitro experiments showed that the recombinant antimicrobial peptide CP is heat and acid-stable, and it has high antibacterial activity against several Gram-positive and -negative bacteria. Only 1 microg of the recombinant antimicrobial peptide CP has an antibacterial activity equivalent to 64 U ampicillin. Thus, this recombinant antimicrobial peptide could serve as an attractive candidate for the development of therapeutic antimicrobial drugs.
Niu
Mingfu
M
Food and Bioengineering College, He'nan University of Science and Technology, Luoyang, He'nan 471003, PR China.
Li
Xiang
X
Wei
Jianchao
J
Cao
Ruibing
R
Zhou
Bin
B
Chen
Puyan
P
D000143:Acids / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects* / Q000235:genetics; D003001:Cloning, Molecular; D015195:Drug Design; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D006358:Hot Temperature; D066298:In Vitro Techniques; D008826:Microbial Sensitivity Tests; D010843:Pichia / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014170:Transformation, Genetic
D000143:Acids; D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins