title: C-terminus engineering of soybean proglycinin: improvement of emulsifying properties.
abstract: Introduction of the extension region of beta-conglycinin alpha' subunit at the C-terminus of proglycinin A1aB1b results in the improvement of its emulsifying properties. To understand the basic for such improvement, we introduced the alpha' and alpha extension regions to the A2B1a C-terminus, and the alpha extension and A5A4B3 hypervariable regions, and an oligopeptide composed of 20 negatively or positively charged residues to the A1aB1b C-terminus, creating A2B1aalpha', A2B1aalpha, and A1aB1balpha, A1aB1bA4IV, A1aB1bNeg and A1aB1bPos, respectively. All the modified versions were produced in Escherichia coli. Their molecular size, thermal stability, surface hydrophobicity, solubility and emulsifying ability were studied. Analyses of molecular size and thermal stability suggested that all the modified versions formed the proper conformation similar to that of the wild type (WT). Solubility was intrinsic to each mutant. At ionic strength 0.5, the emulsifying abilities of all mutants were better than that of the WT except A1aB1bPos and A1aB1bNeg, and at ionic strength 0.08, all mutants especially A1aB1bPos exhibited better emulsifying ability than did the WT. The order of stability of the emulsion at both ionic strengths (0.08 and 0.5) was A1aB1balpha >or= A2B1aalpha > A1aB1balpha' >or= A2B1aalpha' >> A1aB1bPos > A1aB1bA4IV >or= A1aB1bNeg > A1aB1b, A2B1a. These results indicate that the emulsion stability of proglycinin mutants depends on length and hydropathy profile of the polypeptides added to the C-terminus of proglycinin.
authors.affiliation: Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Krisna
authors.initials: K
authors.lastname: Prak
authors.forename: Kazuyo
authors.initials: K
authors.lastname: Nakatani
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Maruyama
authors.forename: Shigeru
authors.initials: S
authors.lastname: Utsumi
mesh_terms: D002152:Calorimetry, Differential Scanning / Q000379:methods; D003720:Densitometry; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008957:Models, Genetic; D008958:Models, Molecular; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D030262:Soybean Proteins / Q000737:chemistry*; D013025:Glycine max / Q000378:metabolism*; D013696:Temperature
chemical_list: D007477:Ions; D010455:Peptides; D030262:Soybean Proteins; C405345:proglycinin A(1a)B(1b), soybean