High performance affinity chromatography of Bacillus neutral proteases.
Bacillus neutral proteases were purified using bacitracin-silica as an affinity medium. Several chromatographic procedures were investigated, including high speed runs on columns with 40- to 60-microns silica particles. The high speed procedure enabled the purification of 4.9 mg of B. subtilis neutral protease directly from 165-ml culture supernatant within 1.5 h. The neutral proteases of B. polymyxa and B. stearothermophilus were also purified. The latter enzyme was further concentrated by a second affinity chromatography step, using Sepharose with glycyl-D-phenylalanine as a ligand. During the purification procedures isopropanol was used to prevent autodigestion of the enzymes.
Eijsink
V G
VG
Department of Genetics, University of Groningen, Haren, The Netherlands.
van den Burg
B
B
Venema
G
G
D000433:1-Propanol / Q000494:pharmacology; D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism
D001426:Bacterial Proteins; D000433:1-Propanol; D008666:Metalloendopeptidases; C071298:bacillolysin