Isolation,Purification And Activity Analysis Of Antimicrobial Peptide SSH19 From Swine Spleen

Antibacterial peptides show a broad spectrum of antibacterial activity,the antibacterial mechanism is unique and complex,difficult to lead to drug resistance.In this study,a new antimicrobial peptide was isolated and purified from porcine spleen,and the antibacterial activity was detected by peptide synthesis.The antibacterial peptides with strong activity were screened and then analyzed by a series of bioinformatics-related software on its physicochemical properties,Transmembrane region,signal peptide,intracellular localization,modification sites and other aspects of the characteristics of the analysis,to develop a new type of antimicrobial drugs to lay the foundation.In this study,the spleen of the spleen was obtained by aseptic technique,and the spleen extract was obtained by tissue homogenate,ultrasonic disruption,acetic acid extraction and high-speed centrifugation.After extraction by cation exchange chromatography and reverse high performance liquid chromatography(RP-HPLC)The crude extract was isolated and purified,Escherichia coli(ATCC8099)and Staphylococcus aureus(ATCC6538)were used as the indicator of the antibacterial activity test to obtain an antibacterial peptide with strong antibacterial activity.The amino acid sequence of the antimicrobial peptide was obtained by MALDI-TOF-MS mass spectrometry and sequence alignment analysis.Synthesis of porcine spleen antimicrobial peptide SSH19 by peptide synthesizer and detection of antimicrobial activity.The results showed that the synthetic antimicrobial peptide SSH19 had excellent bacteriostatic effect on Escherichia coli(ATCC8099)and Staphylococcus aureus(ATCC6538).The data of physicochemical properties,secondary structure,transmembrane region,signal peptide,intracellular localization,glycosylation,ubiquitination,restriction site and hydrophobicity were analyzed by bioinformatics-related software.The results showed that the antibacterial peptide SSH19 had 7 positive charges,and there were two phosphorylation sites,six restriction sites,hydrophilic,and 76% spiral region.