Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia)

This study was performed to compare the total protein and protease content among the whole fruit, flesh, and peel of nine different pear cultivars. Pear proteases were functionally characterized with respect to three enzyme assays. Proteases from pears were further identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in gel activity staining, and matrix assisted laser desorption/ionization-time of flight mass spectrometry analysis. Flesh from Whasan, Nikita, and Hanareum cultivars contained relatively more total protein and protease and showed high enzyme activities, while Chuwhang contained the lowest amount of protein and protease activity. Protease content and enzyme activities found in the pear flesh or whole fruits were two to six times higher than those in the pear peel. Pear cultivars contained one or two protease bands with molecular weights of 36 kDa and/or 38k Da. The larger band was further identified as a cysteine proteinase with 70% homology to the pear cysteine protease f...