(12) United States Patent (10) Patent No.: US 7,741,438 B2 Steele Et Al
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USOO7741438B2 (12) United States Patent (10) Patent No.: US 7,741,438 B2 Steele et al. (45) Date of Patent: Jun. 22, 2010 (54) METHODS AND COMPOSITIONS 6,066,610 A 5/2000 Sramek ...................... 510,284 INVOLVING ENDOPEPTIDASES PEPO2 AND 6,103,277 A 8/2000 Leufstedt et al. ............... 426,8 PEPO3 6,120,809 A 9/2000 Rhodes .......... 426/36 6,127,142 A 10/2000 Harboe et al. ... ... 435/68.1 (75) Inventors: James L. Steele, Middleton, WI (US); 6,139,889 A 10/2000 Guinee et al. ... ... 426/231 Jeffrey R. Broadbent, Smithfield, UT 6,140,078 A 10/2000 Sanders et al. ............. 435/69.1 (US); Vidya R. Sridhar, Portland, OR 6,183,804 B1 2/2001 Moran et al. ................ 426,582 (US) 6,242,036 B1 6/2001 Han et al. 426,582 6,258,390 B1 7/2001 Budtz 426/36 (73) Assignees: Wisconsin Alumni Research 6,270,823 B1 8, 2001 Jolkin ... 426,478 Foundation, Madison, WI (US); Utah 25. R 1939: y et al... 1. State University, Logan, UT (US) 6,335,0404-1-1 B1 1/2002 HolierOOOer et et al. al. ................... 426/34 c - r - 6,399,121 B1 6/2002 Nielsen ....................... 426/37 (*) Notice: Subsyster's 6,401,604 B1 6/2002 Rose et al. ...... ... 99/455 p C. 154(b) bV 904 d 6,410,076 B1 6/2002 Van Der Meulen 426,582 .S.C. 154(b) by ayS. 64 13,568 B1 7/2002 Rietveld ........ 426,582 6,416,797 B1 7/2002 Han et al. ..................... 426/36 (21) Appl. No.: 10/873,427 6,443,379 B2 9/2002 Lassmann ................... 242,178 1-1. 6,455,092 B1 9/2002 Begueria . 426,582 (22) Filed: Jun. 21, 2004 6,458,394 B1 10/2002 Talbott .... 426/36 O O 6,465,033 B2 10/2002 Menninga et al. 426,512 (65) Prior Publication Data 6,468,570 B1 10/2002 Haddad et al. .............. 426,231 US 2005/0281914A1 Dec. 22, 2005 6,475,290 B2 11/2002 Jones ............................ 134.2 6,475,538 B2 11/2002 Thakar et al. ... 426/43 Related U.S. Application Data 6,485,762 B1 1 1/2002 Rizvi et al. .. ... 426/34 6,548,089 B2 4/2003 Holier et al. ... 426/34 (60) Provisional application No. 60/480,536, filed on Jun. 6.551.635 B2 4/2003 Nielsen ....................... 426/37 20, 2003. 6,558,716 B1 5/2003 Smith et al. ................... 426/36 6,572,901 B2 6/2003 Han et al. ..................... 426/36 (51) Int. Cl. 6,686,324 B2 2/2004 Ramirez et al. ............. 510,218 C07K I4/00 (2006.01) (52) U.S. Cl. ........................ 530/350; 530/300; 435/7.1; 435/220 OTHER PUBLICATIONS (58) Field of Classification Search ........... ... None Seffernicket al. (J. Bacteriology, vol. 183, pp. 2405-2410, 2001).* See application file for complete search history. Wells, Biochemistry, vol. 29, pp. 8509-8517, 1990.* Guo et al. (PNAS, vol. 101, No. 25, pp. 9205-9210, 2004.* (56) References Cited Bartels et al., "Accelerated ripening of Gouda cheese. II. Effect of U.S. PATENT DOCUMENTS freeze-shocked Lactobacillus helveticus on proteolysis and flavor development.” Milchwissenschafi, 42(3): 139-144, 1987. 4,001,133 A 1/1977 Sorgenfreiet al. .......... 252,156 Bartels et al., "Accelerated ripening of Gouda cheese. I. 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In particular embodi 5,776,351 A 7, 1998 McGinness et al. ......... 210,748 5,853,786 A 12, 1998 Anbarci et al. ....... ... 426,582 ments of the invention, these methods and compositions 5,888,966 A 3/1999 Larsen et al. .................. 514/2 apply to the cheesemaking process. The invention also con 5.948,459 A 9, 1999 Telford ....................... 426,512 cerns the use of PepO2 and/or PepO3 polypeptides in the 5.988,052 A 1 1/1999 Abler .......................... 99.494 treatment or prevention of celiac sprue or as a food additive. 6,020,324. A 2/2000 Jamas et al. .................. 514/54 6,026,740 A 2/2000 Abler .......................... 99.494 6 Claims, 9 Drawing Sheets US 7,741.438 B2 Page 2 OTHER PUBLICATIONS Kaminogawa et al., “Identification of low molecular weigh peptides in Gouda-type cheese and evidence for the formulation of these Broadbent et al., “Peptide accumulation and bitterness in cheddar peptides from 23 N-terminal residues of Osl-casein by proteinases of cheese made using single-strain Lactococcus lactis starters with dis Streptococcus Cremoris H61.J. Food Sci., 51(5):1253-1264, 1986. tinct proteinase specificities,” J. Dairy Sci., 81:327-337, 1998. Khalid and Marth, “Purification and partial characterization of a Chavagnatetal, "Purification, characterization, cloning and sequenc prolyl-dipeptidyl aminopeptidase from Lactobacillus helveticus ing of the gene encoding oligopeptidase PepO from Streptococcus CNRZ 32.” Appl. Environ. Microbiol., 56(2):381-388, 1990. therrnophilus A.” FEMS Microbiol Lett., 191:79-85, 2000. Khalid et al., “Peptide hydrolases of Lactobaculus helveticus and Chen and Okayama, “High-efficiency transformation of mammalian Lactobacullus delbrueckii ssp. bulgaricus,” J. Dairy Sci., 74:29-45. cells by plasmid DNA.” Mol Cell Biol. 7(8):2745-2752., 1987. 1991. Chen and Steele, "Genetic characterization and physiological role of Kok and De Vos, In: Genetics and Biotechnology of Lactic Acid endopeptidase O from Lactobacillus helveticus CNRZ32.” Appl. Bacteria, Glasson and de Vos (Eds.), Blackie Academic and Profes Environ. Microbiol. 64(9):3411-3415, 1998. sional, Glasgow, 1994. Chen et al., “Identification and characterization of Pep O2 from Kunjiet al., “The proteolytic systems of lactic acid bacteria.” Antonie Lactobacillus helveticus CNRZ32, an enzyme involved in the hydrolysis of a B-casein derived bitter peptide.” J. Anim. Sci., van Leeuwenhoek, 70: 187-221, 1996. 78(Supp. 1):401, 2000. 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