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(A) Production of functional longipain. Longipain was expressed in Pichia pastoris. The purification process was monitored by 12.5% SDS-PAGE. (B) Effect of pH and temperature on longipain activity. Enzyme activity was assayed using Z-Arg-Arg-MCA (Z-R-R-MCA) or Z-Phe-Arg- MCA (Z-F-R-MCA) as the substrate as described in Experimental Procedures. The maximal proteolytic activity was taken as 100% and the relative activity was determined.
