Purification and determination of inhibitory activity of recombinant soyacystatin for surimi application.
A recombinant soyacystatin (r-soyacystatin) was tested for its inhibitory activity against cysteine proteinase of Pacific whiting and its activity was compared to that of egg white cystatin. A recombinant soyacystatin expressed in Escherichia coli was purified to electrophoretic homogeneity using phenyl-Sepharose and DEAE-Sepharose. Native egg white cystatin was purified by using affinity chromatography on CM-papain-Sepharose generated in our lab. Egg white cystatin and soyacystatin were tested for proteinase inhibitory activity against commercial papain and also cathepsin L purified from Pacific whiting muscle. The r-soyacystatin exhibited papain-like protease inhibition activity comparable to that of the egg white cystatin, which could inhibit papain and Pacific whiting cathepsin L. The r-soyacystatin subsequently inhibited the autolytic activity of Pacific whiting surimi.
Akpinar
Ozlem
O
Department of Food Engineering Gaziosmanpasa University--Tasliciftlik, Tokat, Turkey. akpinar99@yahoo.com
An
Haejung
H
D000818:Animals; D001329:Autolysis; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors; D015891:Cystatins / Q000008:administration & dosage / Q000302:isolation & purification* / Q000494:pharmacology; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D004926:Escherichia coli; D005399:Fishes; D009132:Muscles / Q000201:enzymology*; D010206:Papain / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000494:pharmacology
D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D011994:Recombinant Proteins; C100158:cystatin, egg-white; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L; D010206:Papain