Separation of antimicrobial peptides from extracellular matrices

Fangyi Zhu, Purdue University

Abstract

Porcine SIS (Small Intestine Submucosa) and UBS (Urinary Bladder Submucosa) are Extracellular Matrices (ECMs) that are in use or are being tested as scaffolds in tissue engineering. It has been discovered that they resist bacterial infection naturally, which indicates the presence of an antimicrobial peptide. The goals of this study were to isolate and characterize the antimicrobial component and explore the nature of its antimicrobial activity. The ECMs were incubated with acetic acid at 120°C for 30 min and peptide solutions obtained. The effect of acetic acid on antimicrobial activity was identified and discussed. The peptides derived from ECMs were analyzed by HPLC and separated according to molecular weight. These fractions were tested for their antibacterial activity against Escherichia coli and Staphylococcus aureus, which represent Gram-positive and Gram-negative bacteria, respectively, and activity was not found. Rather, the active fraction was found to be adsorbed on the Sephadex G-100 stationary phase and eluted by acetonitrile/water gradient. The sequence of this fraction was determined by tandem mass spectroscopy. The peptide was then chemically synthesized and its antimicrobial activity as well as its interaction with acetic acid as a determinant of its activity was evaluated. Antimicrobial activity for the peptide was observed.

Degree

Ph.D.

Advisors

Ladisch, Purdue University.

Subject Area

Pharmacology|Biomedical research

COinS