Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
1999-05-06
2002-07-16
Carlson, Karen Cochrane (Department: 1653)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C530S350000, C530S300000, C435S006120, C435S069100, C435S320100, C435S252330, C536S023100
Reexamination Certificate
active
06420527
ABSTRACT:
Throughout this specification, various references are identified by author name in parentheses, The citation to the reference corresponding to the identified author can be found in the section entitled References Cited preceding the claims. The references in that section are hereby incorporated by reference,
BACKGROUND OF THE INVENTION
Thaumatin and monellin are flavor active and flavor enhancing proteins. Thaumatin, as shown by thaumatin II, has a wide variety of uses as a flavor enhancing agent. The octapeptide known as “delicious peptide” or “beefy meaty peptide” (BMP) is reported to enhance flavor and produces an umami and sour taste, especially in beef. Monellin is recognized as a potently sweet protein. Recent work indicates that the 19 to 26 amino acid region of thaumatin is a possible active site for sweetness determination. (Slootstra et al. 1995). Substitution of this region with other amino acid sequences, particularly those having some homology with this region of thaumatin and believed to affect taste, may alter the properties of thaumatin to provide new and useful function.
Recent work with monellin indicates that the lysine located at position 4 in the A chain is a highly likely candidate for involvement as a component of the receptor interaction site of monellin. (Suami et al. 1996), Subsitution of this region with other amino acid sequences, particularly those having some homology with this region of monellin and believed to affect taste, may alter the properties of monellin to provide new and useful function.
SUMMARY OF THE INVENTION
The present invention provides new flavor active proteins. Specifically, the new proteins are modified versions of the thaumatin and monellin proteins. In thaumatin II amino acids in the region of 19 to 26 (amino acids 19-26 of SEQ. ID. NO.2), which appears to be a taste active region, are replaced with other amino acid sequences. Most important is the replacement of this region with the sequences for the octapeptide known as “delicious peptide” or “beefy meaty peptide,” i.e. LYS-GLY-ASP-GLU-GLU-SER-LEU-ALA (SEQ. ID. NO. 6), and the sequence that comprises the segment of protein from the fourth to the eleventh amino acid in the A chain of monellin, i.e. LYS-GLY-TYR-GLU-TYR-GLN-LEU-TYR (amino acids 4-11 of SEQ. ID. NO. 4; SEQ ID. NO.11). When four (4) of the 207 amino acids in the thaumatin b protein amino acid sequence were changed, a protein is produced that has a dramatic savory effect when evaluated with a complex salt enhancer such as L-arginine, ammonium chloride, tartaric acid, monopotassium glutamate, or ribotide. In the presence of the salt enhancer, the new protein with the BMP sequence has a beefy, meaty, brothy impression and mouth feel.
This invention further provides a modified monellin protein, either as A and B chains or as a single joined chain, wherein the amino acids in the region of four to eleven of the A chain (or the homologous section of single chain monellin), which appears to be the taste active region, are replaced with other amino acid sequences. Most important is the replacement of this region with the sequences for the octapeptide known as “delicious peptide” or “beefy meaty peptide,” i.e. LYS-GLY-ASP-GLU-GLU-SER-LEU-ALA (SEQ. ID. NO. 6), and the sequence that comprises the segment of protein from the amino acid region of 19 to 26 in thaumatin II, i.e. LYS-GLY-ASP-ALA-ALA-LEU-ASP-ALA (amino acids 19-26 of SEQ. ID. NO. 2; SEQ. ID NO.12).
This invention further provides the DNA sequences encoding the modified thaumatin and monellin proteins and for derivatives of such proteins and the DNA sequences of the derivatives. It also teaches methods for producing these proteins using genetic recombination techniques which can be used in a variety of microorganisms. According to the present invention, these proteins are expressable in yeast transformed by vectors comprising the nucleic acid sequences encoding the proteins. The microorganisms which have been transformed to express these proteins can be used to cultivate cell lines capable of producing the proteins on a large scale, Furthermore, this invention teaches the use of these proteins as flavor additives and enhancers in food.
REFERENCES:
patent: 5221624 (1993-06-01), Blair et al.
patent: 832972 (1998-01-01), None
patent: 0832972 (1998-01-01), None
Aguilar O.M., et al (1987), Nitrogen fixation protein-Rhizobium meliloti,J. Bacteriol, 169:5393-5400, C28379NBRF.
Akusjarvi G., et al (1981), Hexon protein-Human adenovirus 2,Nucleic Acids Res., 9:1-17, HXAD2BNRF.
Alin P., et al (1989), Gluthathione transferase 8, cytosolic-Rat,Biochem.J., 261:531-539, XURT8CNBRF.
Allison L.A., et al (1985), DNA-directed RNA polymerase II 215K polypeptide-Yeast (Saccharomyces cerevisiae),Cell, 42;599-610, RNBY2LNBRF.
Aoki I., et al; (1991). Eosinophil granule major basic protein I precursor-Guinea pig,FEBS Lett., 279:330-334, S13625NBRF.
Aoki I., et al (1991), Eosinophil granule major basic protein 2 precursor-Guinea pig,FEBS Lett., 282:56-60, S15102NBRF.
Au-Young J., et al (1990), Chitin synthase-Imperfect fungus (Candida albicans),Mol.Microbiol., 4:197-207, S11808NBRF.
Baer R., et al (1984), BRRF2 protein-Epstein-Barr virus (strain B95-8),Nature, 310:207-211, QQBE30NBRF.
Begg G.S., et al (1978), Connective-tissue activating peptide III-Human,Proc.Natl.Acad.Sci U.S.A. Biochemistry, 80:765-769, TGHUNBRF
Benfield P.A., et al (1984), Creative Kinase M chain-Rat,J.Biol.Chem., 259:14979-14984 KIRTCMNBRF.
Benfield P.A., et al (1988), Creative kinase (CK)-Rat,Gene, 63:227-243, JT0277NBRF.
Buskin J.N., et al (1985), Creative kinase M chain-Mouse,J.Mol.Evol., 23:334-341, A23590NBRF.
Calabrese L., et al (1989), Superoxide dismutase (Cu-Zn)-Blue shark,FEBS Lett, 50:49-52, S04623NBRF.
Citron, B.A., et al. (1984), Galactokinase-Yeast (Saccharomyces cerevisiae),J.Bacteriol., 158:269-278, KIBYGGNBRF.
Dawson, P.A., et al. (1989), Oxysterol-binding protein-Rabbit,J.Biol.Chem., 264-16798-16803, A34404NBRF.
Ghosh, S., et al (1990), Transcription factor NF-kappaB-Mouse,Cell, 62:1019-1029, ?A35697NBRF.
Gitt, M.A., et al (1985), DNA-directed RNA polymerase sigma chain-Bacillus subtilits, J.Biol.Chem., 20:7178-7185 A22626NBRF.
Gustafson, G., et al (1989), Alpha-a protein-Barley stripe mosaic virus,Virology, 170:370-377, PAVBBSNBRF.
Haas, R.C., et al (1990), Creative kinase precursor, sarcomere-specific, mitochondrial-Human,J.Biol.Chem., 265:6921-6927, A35756NBRF.
Helfman, D.M., et al (1985), Tropomyosin 1, smooth muscle-Chicken,J.Biol.Chem., 259:14136-14143, TMCHS1NBRF.
Herring, B.P., et al (1990), Myosin-light-chain kinase, skeletal muscle-Rabbit,J.Biol.Chem., 265:1724-1730, A35021NBRF.
Hirsch-Behnam, A, et al (1990), Hypothetical protein E1-Human papillomavirus,Virus Res., 18:81-98, S15616NBRF.
Holt, J.C., et al (1986), Platelet basic protein-Human,Biochemistry, 25:1988-1996, A24448NBRF.
Hossle, J.P., et al (1986), B-creatine kinase protein-Chicken,Nucleic Acids Res., 14:1449-1463, A24793NBRF.
Hossle, J.P., et al (1988), Creatine kinase precursor, mitochondrial-Chicken (fragment),Biochem.Biophys.Res.Commun., 51:408-416, A27708NBRF.
Jofuki, K.D., et al (1989), Trypsin inhibitor Kti3+ (Kunitz)-Soybean,Plant Cell, 1:427-435, JQ0968NBRF.
Jornvall, H., et al (1981), Hexon protein-Human adenovirus 2,J.Bioch.Chem., 256:6181-6186, HXAD2NBRF.
Larimer, F.W., et al (1989), revl protein-Yeast (Saccharomyces cerevisiae),J.Bacteriol., 171:230-237, A32240NBRF.
Lehman, L.J., et al (1990), Dinitrogenase reductase-Rhodospirillum rubrum,Gene, 95:143-147, JW0039NBRF.
Levin, D.E., et al (1990), Protein kinase 1-Yeast,Proc.Natl.Acad.Sci.U.S.A., 87:8272-8276, A36474NBRF.
Lin, C.S., et al (1988) .L-plastin-Human,Mol.Cell.Biol., 8:4659-4668, A31559NBRF.
Mariman, E.C.M., et al (1989), Creatine kinase chain B-Human,Nucleic Acids Res., 17-6385, S15935NBRF.
Mariman, E.C.M., et al (1987), Creatine kinase B chain-Human,Genomics, 1:126-137, A27174NBRF.
Michaels, M.L., et al (1990), Adenine glycosylase-Escherichia coli, Nucleic Acids Res., 18:3841-3845, JQ0546BNRF.
Mukai, J, et al (1991), Calcineurin B-like protein-
Bolen Paul L.
Cihak Paul L.
Hawn Regina D.
Kossiakoff Nicolas
Miller Kevin P.
Carlson Karen Cochrane
International Flavors & Fragrances Inc.
Leightner Joseph F.
Robinson Hope A.
LandOfFree
Flavor active modified thaumatin and monellin and methods... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Flavor active modified thaumatin and monellin and methods..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Flavor active modified thaumatin and monellin and methods... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2867053