Fractionation and purification of the thiol proteinases from papaya latex.
Three cysteine proteinases, i.e. chymopapain, papaya proteinase IV and proteinase III, were purified to homogeneity from papaya latex using a combination of ion-exchange chromatography and hydrophobic interaction chromatography. During the purification procedure, the thiol-groups of the active center were reversibly blocked as mixed disulfides with 2-thiopyridone. Homogeneity was proved electrophoretically by native polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulfate (SDS)-PAGE and rechromatography on a Mono S 5/5 column at pH 5.0.
Department of Pharmaceutics, University of Ghent, Belgium.
P M
PM
Dekeyser
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S
De Smedt
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J
Demeester
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A
Lauwers
D000581:Amidohydrolases / Q000032:analysis; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D002917:Chymopapain / Q000302:isolation & purification; D003546:Cysteine Endopeptidases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007840:Latex / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis; D010944:Plants / Q000201:enzymology*; D011480:Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology
D007840:Latex; D010940:Plant Proteins; D011480:Protease Inhibitors; D003546:Cysteine Endopeptidases; C050874:glycyl endopeptidase; C039974:caricain; D002917:Chymopapain; D000581:Amidohydrolases