Potential improvement of the thermal stability of sweet-tasting proteins by structural calculations

Abstract

The low thermal stability of the sweet-tasting proteins limited their applications in food industry. Improve their thermal stability is the key to developing their applications in food processing. In the present study, saturation mutagenesis was performed on 4 sweet-tasting proteins, brazzein (988 mutations), curculin (2109 mutations), monellin (1824 mutations) and thaumatin (3933 mutations), using structural calculations in order to find more thermal stable mutations. The obtained results indicated that our calculated ΔΔG value (ΔΔG < 0 stabilizing, ΔΔG > 0 destabilizing) was a good predictor for predicting changes in thermal stability caused by mutations. Moreover, mutating the negatively charged residues to the other non-negatively charged amino acids was an efficient way to improve the thermal stability of the investigated sweet-tasting proteins. In addition, some promising mutations sites were identified for improving thermal stability using mutagenesis. This study provides useful information for future protein engineering to improve the thermal stability of the sweet-tasting proteins.