Affinity chromatography of proteolytic enzymes on silica-based biospecific sorbents.
New biospecific sorbents for affinity chromatography of proteolytic enzymes were prepared by the attachment of the cyclopeptide antibiotics bacitracin, bacilliquin or gramicidin S to aminosilochrom via a reaction with p-benzoquinone. The content of the cyclopeptide ligands within the sorbents varied from 2 to 46 mumol/g. The sorbents prepared by this reaction were successfully applied in the purification of the carboxylic proteinases produced by fungi, Russula decolorans (a basidiomycete) and Trichoderma lignorum, as well as crude pepsin. Serine proteinases from Thermoactinomyces vulgaris, Trichoderma koningii, Trichoderma lignorum and bacilli (subtilisins) were also submitted to chromatography on these materials. The yields of purified enzymes approached quantitative levels, sometimes being higher as a result of elimination of inhibitors. An important advantage of these sorbents is their stability against the enzymes degrading the carbohydrate matrixes of affinity sorbents synthesized on the basis of agarose, dextran or cellulose derivatives.
D000818:Animals; D001414:Bacitracin; D002846:Chromatography, Affinity / Q000379:methods; D010450:Endopeptidases / Q000302:isolation & purification; D005658:Fungi / Q000201:enzymology; D008848:Micromonosporaceae / Q000201:enzymology; D010434:Pepsin A / Q000302:isolation & purification; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D012697:Serine Endopeptidases; D012822:Silicon Dioxide; D013552:Swine; D014242:Trichoderma / Q000201:enzymology
D001414:Bacitracin; D012822:Silicon Dioxide; D010450:Endopeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D010434:Pepsin A