Recombinant Production of Snakin-2 (an Antimicrobial Peptide from Tomato) in E. coli and Analysis of Its Bioactivity.
Antimicrobial peptides (AMPs) represent a diverse group of biologically active molecules that are part of the innate immune systems of a variety of organisms. Their primary function consists of protecting the host organism against invading microorganisms, including pathogens. AMPs show a broad spectrum of secondary structures, which are essential for antimicrobial activity. In this study, we produced snakin-2 (SN2), a 66-amino-acid-(aa)-long AMP from Solanum lycopersicum as a recombinant protein in E. coli. This AMP belongs to the GASA/GAST protein family and possesses a highly conserved 60-aa-long domain with six disulfide bonds in the C-terminus of the peptide. Because of the toxicity of SN2 against its producing E. coli strain, the AMP was attached to an N-terminal fusion protein (thioredoxin A), which was removed after affinity chromatography purification. The total yield of recombinant SN2 was approximately 1 mg/L. The membrane-active SN2 showed a bactericidal and fungicidal bioactivity, which can be explained by perforation of biomembranes of bacteria and fungi.
Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany. Herbel@uni-heidelberg.de.
Vera
V
Herbel
Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany. Holger.schaefer@uni-heidelberg.de.
Holger
H
Schäfer
Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany. Wink@uni-heidelberg.de.
Michael
M
Wink
D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors / Q000378:metabolism; D018551:Solanum lycopersicum / Q000737:chemistry*; D008826:Microbial Sensitivity Tests; D010940:Plant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology*; D059748:Proteolysis / Q000187:drug effects; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects
D023181:Antimicrobial Cationic Peptides; D010940:Plant Proteins; D011994:Recombinant Proteins
Escherichia coli; antimicrobial peptide; recombinant expression; snakin-2