Introduction of enterostatin (VPDPR) and a related sequence into soybean proglycinin A1aB1b subunit by site-directed mutagenesis.
Enterostatin (VPDPR), having anoretic and hypocholesterolemic activities, and its homologue LPYPR, a hypocholesterolemic peptide found in the glycinin A5A4B3 subunit, were introduced into the corresponding site (TNGPQ) of the proglycinin A1aB1b subunit by site-directed mutagenesis. Modified proglycinins were expressed in E. coli and recovered from the insoluble fraction. VPDPR and LPYPR were released by the action of chymotrypsin and trypsin as expected. The overall yields of purified VPDPR and LPYPR were 40% and 62%, respectively.
Takenaka
Y
Y
Research Institute for Food Science, Kyoto University, Uji, Japan. takenaka@food2.food.kyoto-u.ac.jp
Utsumi
S
S
Yoshikawa
M
M
D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism; D005916:Globulins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D030262:Soybean Proteins; D013025:Glycine max / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D005916:Globulins; D009842:Oligopeptides; D011498:Protein Precursors; D021122:Protein Subunits; D011994:Recombinant Proteins; D030262:Soybean Proteins; C107385:valyl-prolyl-aspartyl-prolyl-arginine; C011474:glycinin; D002918:Chymotrypsin; D014357:Trypsin