Purification of a 'double-headed' inhibitor of alpha-amylase/proteinase K from wheat germ by expanded bed chromatography.
The double-headed inhibitor of alpha-amylase and Proteinase K was purified from wheat germ using Cu(II)-Streamline-chelating resin. The endogenous alpha-amylase could be inactivated by heating. Followed by this step, both packed bed and expanded bed gave similar activity yield of around 83% and fold purification around 23. In the case of expanded bed, it was not necessary to separate precipitated protein before the chromatography. The purified preparation gave a single band on SDS-PAGE and the estimated molecular weight of 21 kDa was in agreement with the reported value for the inhibitor designated as PKI-3 in the literature.
Roy
I
I
Chemistry Department, Indian Institute of Technology, Hauz Khas, New Delhi.
Gupta
M N
MN
D000818:Animals; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D066298:In Vitro Techniques; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D014908:Triticum / Q000737:chemistry; D000516:alpha-Amylases / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D011480:Protease Inhibitors; D000516:alpha-Amylases; D019286:Endopeptidase K