Abstract
Natural sweet protein monellin has a high sweetness and low calorie, suggesting its potential in food applications. However, due to its low heat and acid resistance, the application of monellin is limited. In this study, we show that the thermostability of monellin can be improved with no sweetness decrease by means of sequence, structure analysis, and site-directed mutagenesis. We analyzed residues located in the α-helix as well as an ionizable residue C41. Of the mutants investigated, the effects of E23A and C41A mutants were most remarkable. The former displayed significantly improved thermal stability, while its sweetness was not changed. The mutated protein was stable after 30 min incubation at 85°C. The latter showed increased sweetness and slight improvement of thermostability. Furthermore, we found that most mutants enhancing the thermostability of the protein were distributed at the two ends of α-helix. Molecular biophysics analysis revealed that the state of buried ionizable residues may account for the modulated properties of mutated proteins. Our results prove that the properties of sweet protein monellin can be modified by means of bioinformatics analysis, gene manipulation, and protein modification, highlighting t...Continue Reading
References
Jan 1, 1979·Advances in Protein Chemistry·P L Privalov
May 7, 1976·Analytical Biochemistry·M M Bradford
Sep 1, 1990·Agricultural and Biological Chemistry·M KohmuraY Ariyoshi
Feb 1, 1991·Agricultural and Biological Chemistry·M KohmuraY Ariyoshi
Sep 21, 1992·FEBS Letters·T TancrediP A Temussi
Aug 7, 1990·Biochemistry·K A Dill
Aug 1, 1989·Protein Engineering·S H KimT K Lee
Aug 20, 1987·Nature·C OgataS H Kim
Dec 1, 1973·Acta Physiologica Scandinavica·J N BrouwerY Zotterman
Feb 1, 1995·Chemical Senses·J R SomozaS H Kim
Apr 25, 2000·Archives of Biochemistry and Biophysics·F M Assadi-PorterJ L Markley
Jan 12, 2001·Journal of Molecular Biology·R SpadacciniP A Temussi
Mar 30, 2001·The Journal of Biological Chemistry·Y H SungW Lee
Apr 23, 2003·Journal of Molecular Biology·Roberta SpadacciniPiero A Temussi
Jan 1, 1959·Advances in Protein Chemistry·W KAUZMANN
Sep 24, 2004·Proteins·Wei-Feng XueSara Linse
Apr 1, 2006·Journal of Molecular Biology·Wei-Feng XueSara Linse
Jun 13, 2006·Journal of Molecular Biology·Veronica EspositoPiero A Temussi
Mar 3, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·J R HobbsG L Conn
Jun 4, 2008·Annual Review of Biochemistry·D Wayne Bolen, George D Rose
Dec 23, 2008·Biochimica Et Biophysica Acta·Wei-Feng XueJannette Carey
May 16, 2009·Trends in Biochemical Sciences·Piero Andrea Temussi
Nov 16, 2010·Molecular BioSystems·Olga SzczepankiewiczSara Linse
Feb 24, 2011·Chemical Senses·Catherine M TempletonGraeme L Conn
Jul 29, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Bo LiuMeng Cui
Sep 25, 2012·Biochemical and Biophysical Research Communications·Bo LiuMeng Cui
Oct 24, 2012·Biochemistry·Nilesh AgheraJayant B Udgaonkar
Dec 4, 2014·Food Chemistry·Michele Fortunato RegaDelia Picone
Citations
Oct 25, 2016·FEBS Letters·Roberta SpadacciniDelia Picone
Mar 1, 2018·Frontiers in Molecular Biosciences·Tetsuya MasudaFumito Tani
May 20, 2018·Planta·Daniela CastigliaNunzia Scotti
Apr 4, 2021·Life·Masoud DelfiDelia Picone
Jul 6, 2021·Frontiers in Nutrition·Xiangzhong ZhaoBo Liu