Characterization of proteinases from Antarctic krill (Euphausia superba).
Fractions of three trypsin-like proteinases, TL I, TL II, and TL III, a chymotrypsin-like proteinase, CL, two carboxypeptidase A enzymes, CPA I and CPA II and two carboxypeptidase B enzymes, CPB I and CPB II, from Antarctic krill (Euphausia superba) have been characterized with respect to purity by the means of capillary electrophoresis, CE, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The masses of the trypsin-like and chymotrypsin-like proteinases were determined to be 25,020, 25,070, 25,060, and 26,260Da for TL I, TL II, TL III, and CL, respectively. The masses of the CPA enzymes are likely 23,170 and 23,260Da, whereas the CPB enzyme masses likely are 33,730 and 33,900Da. The degradation efficiency and cleavage pattern of the trypsin-like proteinases were studied with native myoglobin as a model substrate using CE, MALDI-TOF-MS, and nanoelectrospray mass spectrometry (nESI-MS). The degradation efficiency of the trypsin-like proteinases was found to be approximately 12 and 60 times higher compared to bovine trypsin at 37 degrees C and 1-3 degrees C, respectively. All three fractions of trypsin-like proteinases showed a carboxypeptidase activity in combination with their trypsin activity.
D000818:Animals; D000864:Antarctic Regions; D002918:Chymotrypsin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D019075:Electrophoresis, Capillary; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D033201:Euphausiacea / Q000201:enzymology*; D007700:Kinetics; D013058:Mass Spectrometry; D008970:Molecular Weight; D009211:Myoglobin / Q000378:metabolism; D015203:Reproducibility of Results; D013379:Substrate Specificity; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D009211:Myoglobin; D010450:Endopeptidases; D002918:Chymotrypsin; D014357:Trypsin