Protein sweetener

Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues – 25 or more amino acid residues in defined sequence

Reexamination Certificate

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Reexamination Certificate

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07812122

ABSTRACT:
Disclosed herein are sweet proteins that are variants of Brazzein, and nucleotide sequences capable of expressing them. Through a replacement of a tyrosine residue at the C terminus in the naturally occurring Brazzein sequence, or the insertion of two residues (at least one being isoleucine, glycine or proline) before the N terminus of wild type Brazzein, sweetness potency, the taste profile and sweetness strength are improved.

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patent: WO 00/61759 (2000-10-01), None
5 pages of ISR and associated Invitation dated Jan. 9, 2009 in connection with the corresponding PCT/U.S. 2008/055913.
Markley, J., et al.; “Solution Structure of the Thermostable Sweet-Tasting Protein Brazzein”, Nature Structure Biology, vol. 5, No. 6, Jun. 1998, pp. 427-431.
Assadi-Porter, F., et al.; “Efficient Production of Recombinant Brazzein . . . ”, Archives of Biochemistry and Biophysics, vol. 376, No. 2, Apr. 15, 2000, pp. 252-258.
Hellekant, et al.; “Characterization and Chemical Modication of Brazzien . . . ”, Acta Botanica Yunnanica, 1996; 18 (2): 123-133.
Jin, Z., et al.; “Monkey Electrophysiological and Human Psychophysical Responses to Mutants of the Sweet Protein Brazzein . . . ”, Chem. Senses 28: 491-498, 2003.
Jin, Z., et al.; “Critical Regions for the Sweetness of Brazzein”, FEBS Letters 544 (2003 33-37.
Assadi-Porter, F., et al.; “Sweetness Determinant Sites of Brazzein . . . ”, Archives of Biochemistry and Biophysics, vol. 376, No. 2, Apr. 15, 2000, pp. 259-265.
Butt, T. R., et al.; “SUMO fusion technology for difficult-to-express proteins”, Protein Expression and Purification 43 (2005) 1-9.
Izawa, H., et al.; “The Structure-Activity Relationships of the Sweet Protein Brazzein . . . ”, Peptide Science—Present and Future, 1999, pp. 750-751.
Assadi-Porter et al. J. Mol. Biol., 2010—article in press, “Key Amino Acid Residues Involved in Multi-Point Binding Interactions between Brazzein, a Sweet Protein, and the T1R2-T1R3 Human Sweet Receptor”.
8 pages of an international preliminary report dated Sep. 15, 2009 in connection with the corresponding PCT/U.S. 2008/055913.

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