1.
Protein Expr Purif
; 96: 26-31, 2014 Apr.
Artigo
em Inglês
| MEDLINE
| ID: mdl-24492011
RESUMO
A winged bean trypsin inhibitor (WbTI-2) of molecular mass â¼20kDa, has been cloned and expressed in Escherichiacoli with full activity like the one from seed protein. It completely inhibits trypsin at an enzyme:inhibitor molar ratio of 1:2. PCR with cDNA and genomic DNA using same primers produced about 550 base pair product, which indicated it to be an intronless gene. Through site-directed mutagenesis, the Arg64 has been confirmed as the P1 residue. For the presence of five methionine residues in WbTI-2, cyanogen bromide (CNBr) digestion was carried out. Out of three fragments the one (about 65% of original size) containing the reactive site loop retained 50% activity.