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Affinity chromatography of proteinases using bacitracin immobilized to porous glass beads
Author(s) -
Fontecha J.,
Requena T.,
Swaisgood H.E.
Publication year - 1996
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1996.tb01181.x
Subject(s) - bacitracin , autolysis (biology) , chromatography , affinity chromatography , porous glass , chemistry , adsorption , proteinase k , enzyme , biochemistry , porosity , organic chemistry , antibiotics
J. FONTECHA, T. REQUENA AND H.E. SWAISGOOD. 1996. This study describes an affinity chromatography procedure for proteinase purification using bioselective binding to immobilized bacitracin. By coupling bacitracin to controlled‐pore glass (CPG) beads, an affinity matrix was obtained that permitted rapid purification of proteinases under conditions that minimize autolysis. Bacitracin‐CPG was used to bioselectively adsorb the extracellular proteinase secreted by Enterococcus faecalis var. liquefaciens IFPL 383. The overall purification obtained with this procedure was 5149‐fold. The ability of bacitracin‐CPG to bind other proteinases was examined using various commercial proteinases. The specific activities of subtilin BPN' and proteinase K were increased by bioselective adsorption and excellent recoveries of all proteinases applied were obtained.