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US 20140134292A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0134292 A1 Petersen (43) Pub. Date: May 15, 2014 (54) PRODUCTION OF CHEESE WITH S. Publication Classi?cation THERMOPHILUS (51) Int. Cl. (75) Inventor: Lars Wexoe Petersen, Muskego, WI A23C 19/032 (2006.01) (US) A23C 19/076 (2006.01) C12R 1/46 (2006.01) (73) Assignee: DUPONT NUTRITION (52) us CL BIOSCIENCES APS, COPENHAGEN CPC ............... .. A23C 19/032 (2013.01); C12R 1/46 (DK) (2013.01); A23C 19/076 (2013.01) USPC ......................... .. 426/36; 435/253.4; 426/582 (21) Appl. No.: 14/112,892 (22) PCT Filed: Jan. 12, 2012 (57) ABSTRACT (86) PCT No.1 PCT/U512/21113 The present invention provides methods, compositions, and 371 1 systems for producing cheese With S. lhermophilus and a § (6X )’_ urease inhibitor, and for producing cottage cheese With S. (2), (4) Date- Jan“ 7’ 2014 Zhermophilus that is partially or completely de?cient in its . ability to release ammonia from urea. The present invention Related U's' Apphcatlon Data also provides methods, compositions, and systems for reduc (60) Provisional application No. 61/477,211, ?led on Apr. ing the amount of open texture (e. g., slits, cracks, or fractures) 20, 2011. in gassy cheeses, such as cheddar cheese. Patent Application Publication M y 15, 2014 Sheet 1 0f 3 US 2014/0134292 A1 Activity Pro?le at 35°C 1% fresh mllk (‘0)Temp Time (mins) - - - Ladococci (570 g) and ur(+) S. thermophilus (140 g) - i- — Lac'bococci (570 g) and ur(+) S. thermophilus (140 g) —B——Lactococci (640 g) and ur(-)S. mes-mophilus (709) —-—¢—Lactocucci (640 g) and ur(-) Sv thermophilus (70 g) - Milk Temp (1:) Figure 1 Patent Application Publication May 15, 2014 Sheet 2 0f 3 US 2014/0134292 A1 Activity Profile at 35‘C 2% fresh milk Time (mins) ur(+) Si ihermophiius with sodium formate (10 ppm) _ur(<) S. thermophilus with sodium formaie (1 0 ppm) - - v - ur(—) S. ihermophilus Wilhuut sodium formats Milk Temp (‘0) Figure 2 Patent Application Publication May 15, 2014 Sheet 3 0f3 US 2014/0134292 A1 Figure 3 US 2014/0134292 A1 May 15, 2014 PRODUCTION OF CHEESE WITH S. the presence of urea, such as the equipment being tied up for THERMOPHILUS a longer time, increased risk of contamination, high ammonia content of the whey, etc. FIELD [0007] US. Pat. No. 6,962,721, which is hereby incorpo rated by reference in its entirety, describes the use of Strep [0001] In one aspect, methods, compositions, and systems tococcus thermophilus strains lacking the ability, or having for producing cheese with S. thermophilus and a urease reduced ability, to hydrolyze urea, (herein termed S. thermo inhibitor, and for producing cottage cheese with S. thermo philus “ur(—) bacteria”) as lactic ferrnents in the production of philus that is partially or completely de?cient in its ability to dairy products. The inventors have unexpectedly found that release ammonia from urea are provided. Methods, compo many of the above-mentioned problems can be resolved by sitions, and systems for reducing the amount of open texture using ur(—) Streptococcus thermophilus bacteria. (e. g., slits, cracks, or fractures) in gassy cheeses, such as, for example, cheddar cheese are also provided. BRIEF DESCRIPTION OF THE FIGURES [0008] FIG. 1 is a graph with a table insert showing exem BACKGROUND plary activity pro?les of ur(+) and ur(—) bacteria; [0009] FIG. 2 is a graph showing exemplary activity pro [0002] Streptococcus thermophilus is a thermophilic lactic bacterium used as a lactic ferment in the dairy industry. First ?les of ur(+) and ur(—) bacteria; and used for the manufacture of fermented milks such as yoghurt, [0010] FIG. 3 is a photograph showing an exemplary result it is now increasingly used in cheese production, for example, from a ?oating curd experiment in test tubes. in production of cheeses that was formerly made with Lacto SUMMARY cocci bacteria, such a Lactococcus lactis or Lactococcus cre morzs. [0011] Methods, compositions, and systems for producing [0003] This bacterium converts lactose in milk into lactic cheese with S. thermophilus and a urease inhibitor, and for acid, which acidi?es the milk. In the case of cheeses, this producing cottage cheese with S. thermophilus that are par acidi?cation not only encourages the action of the rennet and tially or completely de?cient in their ability to release ammo the synaeresis of the curds, but also inhibits the growth of nia from urea are provided. Methods, compositions, and sys many undesirable bacteria, certain of which are pathogenic tems for reducing the amount of open texture (e.g., slits, bacteria, and allows their elimination at a greater or lesser cracks, or fractures) in gassy cheeses, which may include speed. cheeses that produce gas during ripening, such as, for example, cheddar cheese, are also provided. [0004] The acidifying activity of this bacterium is accom panied by urea hydrolysis activity, which affects the acidi? [0012] Various exemplary bacterial strains are occasionally cation kinetics. Tinson et al (1982) showed that the urea referred to herein. Certain strains are referred to by the hydrolysis reaction, which converts urea into carbon dioxide nomenclature CNCM followed by letters and/or numbers, or and ammonia, results in a temporary decrease in the acidi? DSM followed by letters and/or numbers. These references cation speed, as measured by a pH probe. are the deposit numbers at the Collection Nationale de Cul tures de Microorganismes (CNCM) and the Deutsche Sam [0005] On an industrial scale, the hydrolysis of urea by mlung von Mikroorganismen (DSMZ), respectively. All Streptococcus thermophilus poses a number of problems. strains referred to by such numbers have been deposited in the This is because, in cheese manufacturing for example, the respective culture depositories under the reference numbers technological operations (cutting of the curds, stirring, etc.) referred to herein, as follows: CNCM 1-231 1 was deposited at must take place at given values of pH, but in practice these the CNCM on 14 Sep. 1999 by Texel/Rhodia services and is operations are generally carried out at predetermined times. described in US. Pat. No. 6,962,721 which is hereby incor Therefore the variations in acidifying activity due to urea porated by reference it its entirety; CNCM 1 -23 12 was depos hydrolysis lead to defects and signi?cant variability in the ited at the CNCM on 14 Sep. 1999 by Texel/Rhodia services texture, moisture level, and ripening properties of the result and is described in US. Pat. No. 6,962,721 which is hereby ing cheeses. Moreover, because ammonia is basic, the pro incorporated by reference in its entirety; CNCM 1-2980 was duction of ammonia increases the time necessary to reach a deposited at the CNCM on 26 Feb. 2003 by Rhodia Food given pH. This results in the cheese-making equipment being SAS, and is described in WO 04/085607which is hereby tied up for longer and in an increase of the risk of contami incorporated by reference in its entirety; CNCM 1-3617 was nation by undesirable micro-organisms. Furthermore, it is deposited at the CNCM on 14 Jun. 2006 in the name of desirable that the cheese-making whey does not contain an Danisco France SAS and is described in WO 08/040734 excessive amount of ammonia, because this whey is often which is hereby incorporated by reference in its entirety; used as an ingredient in human food and animal feed. The DSM 21892 was deposited at the DSMZ on 7 Oct. 2008 in the production of ammonia from urea is dif?cult to control, in name of Danisco Deutschland GmbH and is described in WO part because the urea content of milk is variable (for example, 10/066907 which is hereby incorporated by reference in its from 2 to 8 mM) and depends in part on the diet of the entirety; and DSM 18344 was deposited at the DSMZ on 14 livestock that produce the milk. Jun. 2006 and is described in WO 07/144770 which is hereby [0006] To overcome this problem, Martin et al (1997) pro incorporated by reference in its entirety. posed measuring the urea content of the milk and then adapt [0013] In one aspect, methods for producing cheese, such ing the manufacturing parameters. However, such a system, as cottage cheese, are provided comprising the following which requires quantitatively determining the amount of steps: a) inoculating milk with ur(—) Streptococcus thermo urea, would be highly constraining, and would not resolve the philus bacteria, wherein the S. thermophilus bacteria are not other drawbacks caused by reduction of acidi?cation speed in able to release ammonia from urea, or wherein the S. thermo US 2014/0134292 A1 May 15, 2014 philus bacteria have a diminished ability to release ammonia consisting of: CNCM 1-2980, DSM21892, CNCM 1-3617, from urea compared to wild-type S. thermophilus; b) fer CNCM 1-3617, CHCC4325, DSM18344, and DSM181 1 1, in menting the milk with the ur(—) Streptococcus thermophilus a process for producing cottage cheese. bacteria; and c) optionally making further adequate steps [0021] Particular aspects provide methods for producing a resulting in the produced cheese, which in some aspects is dairy product such as cheese (e.g., cottage cheese, cheddar cottage cheese. See, e.g., methods of making cottage cheese cheese, mozzarella, pizza cheese, blue cheese, Swiss cheese, in Us. Pat. Nos. 6,482,460; 6,238,717; 3,298,836; WO91/ or any other type of cheese) or yogurt comprising: a) inocu 00690; and Us. Pat. No. 3,968,256; all ofwhich are hereby lating milk with Streptococcus thermophilus bacteria and a incorporated by reference in their entirety.