ISOLATION AND PRELIMINARY CHARACTERIZATION OF THE CYSTEINE-PROTEINASES FROM THE LATEX OF CARICA-CANDAMARCENSIS HOOK

The cysteine-proteinase chymopapain from Carica papaya L. is used for chemonucleolysis of damaged human intervertebral spinal discs. The pur ification of this enzyme is difficult. To overcome these problems, we were looking for a substitute among the cysteine-proteinases of Carica candamarcensis Hook. The latex from unripe fruits was collected in an aqueous solution of methylethanethiolsulfonate to prevent proteolytic activities. The soluble fraction of the lypophilized product provided four enzymatically active peaks (CC-I-CC-IV) during chromatography on CM-Sephadex C-50 in sodium acetate buffer, pH 5.0. They could be furt her purified by rechromatography under similar conditions. The isolate d enzymes have been characterized by PAGE, analysis of the Fourier tra nsform infrared spectra, preliminary studies of their specificities as well as a comparison of the N-terminal amino-acid sequences up to pos ition 43. CC-III proved to be glycosylated. CC-I and CC-III from Caric a candamarcensis Hook are suggested to correspond to papain and chymop apain from Carica papaya L., respectively.