Evaluation of the solubility and emulsifying property of soybean proglycinin and rapeseed procruciferin in relation to structure modified by protein engineering.
The presence or absence of a highly negatively charged extension region in beta-conglycinin (J. Agric. Food Chem. 1999, 47, 5278) and the length of a highly negatively charged variable region IV in glycinin (J. Agric. Food Chem. 2004, 52, 8197) are important determinants of solubility and emulsifying property. To examine the effects of the variable region IV from proglycinin A1aB1b and A3B4 and of the extension region from beta-conglycinin alpha' (alpha'ext) on solubility and emulsifying properties in detail, several mutants of proglycinin, procruciferin, and beta-conglycinin were designed and prepared in Escherichia coli. Nine out of 10 mutants were expressed at high levels in E. coli and shown to be homotrimer similar to the wild types as assessed by gel filtration. The position of the introduced negatively charged region as well as the amino acid composition were demonstrated to affect solubility at mu = 0.08. All of the proglycinin, procruciferin, and beta-conglycinin mutants with the alpha'ext in the C-terminus, especially the proglycinin mutant, exhibited excellent emulsifying ability and emulsion stability. These indicate that improvement of emulsifying properties by insertion of the alpha'ext in the C-terminus may be generally applicable to seed globulins.
Tandang
Mary Rose G
MR
Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
Atsuta
Nami
N
Maruyama
Nobuyuki
N
Adachi
Motoyasu
M
Utsumi
Shigeru
S
D052179:Antigens, Plant; D029741:Brassica rapa / Q000737:chemistry*; D045703:Emulsifying Agents; D005916:Globulins / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D015202:Protein Engineering; D055314:Seed Storage Proteins; D012995:Solubility; D030262:Soybean Proteins / Q000737:chemistry / Q000235:genetics; D013025:Glycine max / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014162:Transfection
D052179:Antigens, Plant; D045703:Emulsifying Agents; D005916:Globulins; D010940:Plant Proteins; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; C405345:proglycinin A(1a)B(1b), soybean; C011474:glycinin