Exploiting the unique biophysical properties of bacteriocins to purify Bac 1829 from Staphylococcus aureus KSI1829.
Bac1829 from Staphylococcus aureus KSI1829 is a newly identified peptide bacteriocin that inhibits a broad spectrum of bacteria. By taking advantage of its cationic and hydrophobic nature, a purification scheme was developed utilizing preparative isoelectric focusing and hydrophobic interaction chromatography. Due to a high pI value of approximately 9-10, 71% of the total Bac1829 activity was concentrated in two fractions by preparative isoelectric focusing. Final purification by high performance liquid chromatography on a propyl hydrophobic interaction column resulted in a 136-fold purification with an increase in specific activity from 181 to 24,623 antimicrobial units (AU)/mg protein. A total of 14,848 AU of Bac 1829 were purified using this revised procedure, compared to 8702 AU as described previously. Mass spectrographic analysis of the purified sample yielded a single peak of 6418 +/- 2 daltons. Since many bacteriocins possess similar biophysical attributes, the purification scheme presented in this communication may be applicable to those which have proven difficult to purify or have not been purified.
Crupper
S S
SS
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506, USA.
Iandolo
J J
JJ
D001430:Bacteriocins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D005590:Fractional Precipitation; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D013211:Staphylococcus aureus / Q000737:chemistry*
D001430:Bacteriocins; C017724:staphylococcin