β-Conglycinin and glycinin are soybean major seed storage proteins. Previous studies have shown that adding the extension region of β-conglycinin α subunit improves the emulsifying properties of proglycinin and confers more favourable characteristics than fusing the extension region of β-conglycinin α′ subunit or the hypervariable regions (A4IV) of glycinin A1aB1b subunit. To evaluate the polypeptide properties, we designed mutants of A1aB1b subunits fused with truncated versions of A4IV (A4IVcut), α (αcut) or α′ (α′cut) extension regions lacking the C-terminus 25 or 31 residues (A4IVC25, αC25 or α′C31), and also A4IVcut and α′cut with αC25 residues added (A4IVcut-αC25 and α′cut-αC25). All the modified proteins displayed conformations similar to the wild type. With good solubilities, the emulsion properties of the modified proteins were much better at ionic strength μ = 0.08 than at μ = 0.5. The modified A1aB1bαcut and A1aB1bα′cut showed poorer emulsion properties than those of A1aB1bα and A1aB1bα′. Replacing the hydrophobic A4IVC25 region of A1aB1bA4IV with hydrophilic αC25 created A1aB1bA4IVcut-αC25, which had the best emulsion stability among these proglycinin mutants. We found that addition of αC25 improves the emulsifying properties of two C-terminally truncated proglycinin variants, thereby illustrating its potential general utility. Our investigation showed that in order to improve the emulsifying ability and emulsion stability of a globular protein, the introduced polypeptide should (i) be highly hydrophilic, (ii) consist of multiple hydrophobic-strong hydrophilic regions comprising at least two alpha helixes, (iii) harbour a terminal α-helix at the end of the C-terminus and (iv) have properties similar to those of αC25.
CITATION STYLE
Prak, K., Naka, M., Tandang-Silvas, M. R. G., Kriston-Vizi, J., Maruyama, N., & Utsumi, S. (2015). Polypeptide modification: An improved proglycinin design to stabilise oil-in-water emulsions. Protein Engineering, Design and Selection, 28(9), 281–291. https://doi.org/10.1093/protein/gzv031
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