Partial purification and characterization of cysteine proteinase inhibitor from chicken plasma

Purification and Characterization of a Cysteine Protease Inhibitor from Chum Salmon ( Oncorhynchus keta) Plasma

“…Contrarily, no inhibitory zones were detected on the gel under reducing conditions regardless of CSP loading amounts (Figure 1, lanes 6 and 7). Similar results were found during the inhibitory activity staining of CPI from chicken plasma (13). It was suggested that the active form of CSP CPI was possibly stabilized by a disulfide bond, which would be further discussed after CPI was purified from CSP.…”

“…The single step purification in this study resulted in higher yield than that of CPI purification from glassfish egg, which was only 0.25% . The purity of CPI in chicken plasma increased about 25-fold after purification by carboxymethyl- (CM-) papain−Sepharose 4B chromatography , which was a little lower than 30.36-fold in this study. A kininogen was even purified from PPP by a series of DEAE-Sepharose, CM-Sepharose, and Sephacryl S-200 chromatographies .…”

“…Seventy percent of inhibitory activity of CSP CPI was still retained after a 30 min incubation at 50 °C. In contrast, the chum salmon egg inhibitor presented stability only at temperatures lower than 35 °C (15), and partially purified CPI from chicken plasma presented stability after heating for only 10 min (13). Inhibitory activity of CSP CPI decreased to 45% and 18% when incubated at 60 and 70 °C for 30 min, respectively.…”

“…A CPI was purified from CSP by one step of affinity chromatography on CNBr−papain−Sepharose 4B (Table ). Affinity chromatography was widely used in the purification of CPIs from a lot of resources due to its simplicity and high purification capacity – ,,– . Because there are more than 100 kinds of proteins in plasma , , two washing buffers in which the second contained 1 M NaCl were used to remove the unabsorbed and nonspecifically bound proteins.…”

“…Chicken plasma protein (CPP) was applied to bigeye snapper and lizardfish surimi to prevent gel weakening , . A cysteine protease inhibitor (CPI) with the molecular mass of 122 kDa was purified from chicken plasma .…”