Effects of designed sulfhydryl groups and disulfide bonds into soybean proglycinin on its structural stability and heat-induced gelation.
The gel-forming ability of glycinin is one of soybean's most important functional properties. The proglycinin A1aB1b homotrimer was engineered to introduce sulfhydryl groups and disulfide bonds, and their effects on the structural stability and the heat-induced gelation were evaluated. On the basis of the crystal structure, five mutants were designed and prepared: R161C and F163C forming an interprotomer disulfide bond with the inherent free cysteine residue of Cys377, N116C/P248C forming a new intraprotomer disulfide bond, and N116C and P248C introducing a new sulfhydryl group. Mutants of R161C, F163C, and N116C/P248C formed a new disulfide bond as expected. N116C/P248C was significantly more stable than the wild type against chemical and thermal denaturation and more resistant to alpha-chymotrypsin digestion, whereas F163C showed significantly increased thermal stability. All mutants exhibited greater hardness of heat-induced gels than wild type, and in particular, N116C/P248C gave the hardest gel. This result indicates that it is possible to increase hardness of glycinin gel by introduction of cysteine residues using protein engineering.
Adachi
Motoyasu
M
Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Chunying
Ho
H
Utsumi
Shigeru
S
D004220:Disulfides / Q000737:chemistry*; D004355:Drug Stability; D005782:Gels / Q000737:chemistry*; D005916:Globulins / Q000737:chemistry; D006358:Hot Temperature; D008958:Models, Molecular; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D030262:Soybean Proteins / Q000737:chemistry* / Q000235:genetics; D013025:Glycine max / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D004220:Disulfides; D005782:Gels; D005916:Globulins; D030262:Soybean Proteins; D013438:Sulfhydryl Compounds; C405345:proglycinin A(1a)B(1b), soybean; C011474:glycinin