PROCESS FOR THE PREPARATION OF CHEESE

Abstract

The present invention provides a process for the preparation of a cheese, the process comprising the steps of (i) providing milk; (ii) acidifying the milk, where the mild is acidified with carbon dioxide to a pH of 6.2 to 6.6 when measured at a temperature of 29 to 38 C. to provide an acidified milk; (iii) inoculating the acidified milk with a starter culture, wherein the inoculation is a direct vat inoculation, to provide an inoculated acidified milk and making the cheese therefrom; wherein the inoculated acidified milk contains a coagulant, wherein the coagulant comprises at least a microbial protease coagulating agent.

Claims

1. A process for the preparation of a cheese, wherein: the process comprises: (i) providing milk, (ii) acidifying the milk, wherein the milk is acidified with carbon dioxide to a pH of 6.2 to 6.6 when measured at a temperature of 29 to 38 C. to provide an acidified milk, and (iii) inoculating the acidified milk with a starter culture to provide an inoculated acidified milk and making the cheese therefrom; the inoculation is a direct vat inoculation; the inoculated acidified milk contains a coagulant; and the coagulant comprises at least a microbial protease coagulating agent.

2. A process according to claim 1, wherein the cheese is selected from semi-hard cheeses, continental cheeses, gouda cheese, edam cheese, masdam cheese, cheddar cheese, cottage cheese, pasta filata cheese, mozzarella cheese, pizza cheese, hard cheeses, parmesan cheese, swiss type cheese, emmental type cheese, soft cheeses, Tvarog cheese, and lactic curd.

3. A process according to claim 1, wherein the milk is pasteurised milk.

4. A process according to claim 1, wherein the milk is selected from cow milk, sheep milk and goat milk.

5. A process according to claim 1, wherein the milk is selected from whole milk, reconstituted skim milk powder, skim milk, and semi-skim milk.

6. A process according to claim 1, wherein the milk is standardised, homogenised or standardised and homogenised.

7. A process according to claim 1, wherein the milk is acidified by the addition of solid carbon dioxide to the milk, by the bubbling of gaseous carbon dioxide through the milk or a combination thereof.

8. A process according to claim 1, wherein the milk is acidified with carbon dioxide to a pH of 6.3 to 6.6 when measured at a temperature of 29 to 38 C.

9-15. (canceled)

16. A process according to claim 1, wherein the milk is acidified solely with carbon dioxide.

17. A process according to claim 1, wherein the milk is acidified in part with carbon dioxide and in part with at least a secondary acidifying agent.

18. A process according to claim 17, wherein the secondary acidifying agent is selected from a culture, an organic, an inorganic acid, and combinations thereof.

19. A process according to claim 1, wherein the coagulant consists of or consists essentially of a microbial protease coagulating agent.

20. A process according to claim 1, wherein the coagulant comprises a microbial protease coagulating agent and a secondary coagulating agent.

21. (canceled)

22. A process according to claim 1, wherein the microbial protease coagulating agent is a single enzyme coagulating agent.

23. A process according to claim 1, wherein the microbial protease coagulating agent has k-casein cleaving activity.

24. A process according to claim 1, wherein the microbial protease coagulating agent predominantly has k-casein cleaving activity.

25. A process according to claim 1, wherein the microbial protease coagulating agent solely has k-casein cleaving activity.

26. A process according to claim 1, wherein the microbial protease coagulating agent is k-casein specific cleaving enzyme.

27. A process according to claim 1, wherein the microbial protease coagulating agent has pepsin activity.

28. A process according to claim 1, wherein the microbial protease coagulating agent predominantly has pepsin activity.

29. A process according to claim 1, wherein the microbial protease coagulating agent solely has pepsin activity.

30. A process according to claim 1, wherein the microbial protease coagulating agent is a pepsin.

31. A process according to claim 1, wherein the microbial protease coagulating agent has mucorpepsin (EC 3.4.23.23) activity.

32. A process according to claim 1, wherein the microbial protease coagulating agent predominantly has mucorpepsin (EC 3.4.23.23) activity.

33. A process according to claim 1, wherein the microbial protease coagulating agent solely has mucorpepsin (EC 3.4.23.23) activity.

34. A process according to claim 1, wherein the microbial protease coagulating agent is a mucorpepsin (EC 3.4.23.23).

35. A process according to claim 1, wherein the microbial protease coagulating agent is a single enzyme coagulating agent consisting of a mucorpepsin (EC 3.4.23.23) having k-casein specific cleaving activity.

36. A process according to claim 1, wherein the microbial protease coagulating agent comprises mucorpepsin (EC 3.4.23.23) MARZYME.

37. A process according to claim 1, wherein the microbial protease coagulating agent is solely mucorpepsin (EC 3.4.23.23) MARZYME.

38. A process according to claim 1, further comprising the step of measuring the pH of the milk of step (i), and adding carbon dioxide to the milk in amount required to provide acidified milk having a pH of 6.2 to 6.6 when measured at a temperature of 29 to 38 C.

39. A process according to claim 38, wherein the amount of carbon dioxide added is 90 grams [0.2 lbs] of carbon dioxide per 1000 lbs of milk per 0.08 unit pH reduction.

40-43. (canceled)

44. A process according to claim 1, wherein no calcium chloride is added during the process.

Description

DETAILED DESCRIPTION

[0023] As discussed herein, the present invention provides a process for the preparation of a cheese, the process comprising the steps of

(i) providing milk;
(ii) acidifying the milk, wherein the milk is acidified with carbon dioxide to a pH of 6.2 to 6.6 when measured at a temperature of 29 to 38 C. to provide an acidified milk;
(iii) inoculating the acidified milk with a starter culture, wherein the inoculation is a direct vat inoculation, to provide an inoculated acidified milk and making the cheese therefrom;
wherein the inoculated acidified milk contains a coagulant, wherein the coagulant comprises at least a microbial protease coagulating agent.

[0024] Cheese

[0025] The present process may be used to prepare any cheese. In one aspect the cheese is selected from Abbaye de Belloc, Abbaye de Citeaux, Abbaye du Mont des Cats, Abertam, Abondance, Acapella, Ackawi, Acorn, Adelost, Affidelice au Chablis, Afuega'l Pitu, Airag, Airedale, Aisy Cendre, Allgauer Emmentaler, Alverca, Ambert, American Cheese, Ami du Chambertin, Anejo Enchilado, Anneau du Vic-Bilh, Anthoriro, Appenzell, Aragon, Ardi Gasna, Ardrahan, Armenian String, Aromes au Gene de Marc, Asadero, Asiago, Aubisque Pyrenees, Autun, Avaxtskyr, Baby Swiss, Babybel, Baguette Laonnaise, Bakers, Baladi, Balaton, Bandal, Banon, Barry's Bay Cheddar, Basing, Basket Cheese, Bath Cheese, Bavarian Bergkase, Baylough, Beaufort, Beauvoorde, Beenleigh Blue, Beer Cheese, Bel Paese, Bergader, Bergere Bleue, Berkswell, Bethmale des Pyrnes, Bethmale of the Pyrenees, Beyaz Peynir, Bierkase, Bishop Kennedy, Blarney, Bleu d'Auvergne, Bleu de Gex, Bleu de Laqueuille, Bleu de Septmoncel, Bleu de Termignon Alpage, Bleu Des Causses, Blue, Blue Castello, Blue of Termignon, Blue Rathgore, Blue Vein (Australian), Blue Vein Cheeses, Bocconcini, Bocconcini (Australian), Boeren Leidenkaas, Bonchester, Bosworth, Bougon, Boule Du Roves, Boulette d'Avesnes, Boursault, Boursin, Bouyssou, Bra, Braudostur, Breakfast Cheese, Brebis du Lavort, Brebis du Lochois, Brebis du Puyfaucon, Bresse Bleu, Brick, Brie, Brie au poivre, Brie de Meaux, Brie de Melun, Brie with pepper, Brillat-Savarin, Brin, Brin d'Amour, Brin d'Amour, Brinza (Burduf Brinza), Briquette de Brebis, Briquette du Forez, Broccio, Broccio Demi-Affine, Brousse du Rove, Bruder Basil, Brusselae Kaas (Fromage de Bruxelles), Bryndza, Buchette d'Anjou, Buffalo, Burgos, Butte, Butterkase, Button (Innes), Buxton Blue, Cabecou, Caboc, Cabrales, Cachaille, Caciocavallo, Caciotta, Caerphilly, Cairnsmore, Calenzana, Cambazola, Camembert de Normandie, Canadian Cheddar, Canestrato, Cantal, Caprice des Dieux, Capricorn Goat, Capriole Banon, Caravane, Carre de I'Est, Casciotta di Urbino, Cashel Blue, Castellano, Castelleno, Castelmagno, Castelo Branco, Castigliano, Cathelain, Celtic Promise, Cendre d'Olivet, Cerney, Chabichou, Chabichou du Poitou, Chabis de Gatine, Chaource, Charolais, Chaumes, Cheddar, Cheddar Clothbound, Cheshire, Chevres, Chevrotin des Aravis, Chontaleno, Civray, Coeur de Camembert au Calvados, Coeur de Chevre, Cojack, Colby, Colby-Jack, Cold Pack, Comte, Coolea, Cooleney, Coquetdale, Corleggy, Cornish Pepper, Cotherstone, Cotija, Cottage Cheese, Cottage Cheese (Australian), Cougar Gold, Coulommiers, Coverdale, Crayeux de Roncq, Cream Cheese, Cream Havarti, Crema Agria, Crema Mexicana, Creme Fraiche, Crescenza, Croghan, Crottin de Chavignol, Crottin du Chavignol, Crowdie, Crowley, Cuajada, Curd, Cure Nantais, Curworthy, Cwmtawe Pecorino, Cypress Grove Chevre, Danablu (Danish Blue), Danbo, Danish Fontina, Daralagjazsky, Dauphin, Delice des Fiouves, Denhany Dorset Drum, Derby, Dessertnyj Belyj, Devon Blue, Devon Garland, Dolcelatte, Doolin, Doppelrhamstufel, Dorset Blue Vinney, Double Gloucester, Double Worcester, Dreux a la Feuille, Dry Jack, Duddleswell, Dunbarra, Dunlop, Dunsyre Blue, Duroblando, Durrus, Dutch Mimolette (Commissiekaas), Edam, Edelpilz, Emental Grand Cru, Emlett, Emmental, Epoisses de Bourgogne, Esbareich, Esrom, Etorki, Evansdale Farmhouse Brie, Evora De L'Alentejo, Exmoor Blue, Explorateur, Farmer, Feta, Feta (Australian), Figue, Filetta, Fin-de-Siecle, Finlandia Swiss, Finn, Fiore Sardo, Fleur du Maquis, Flor de Guia, Flower Marie, Folded, Folded cheese with mint, Fondant de Brebis, Fontainebleau, Fontal, Fontina Val d'Aosta, Formaggio di capra, Fougerus, Four Herb Gouda, Fourme d'Ambert, Fourme de Haute Loire, Fourme de Montbrison, Fresh Jack, Fresh Mozzarella, Fresh Ricotta, Fresh Truffles, Fribourgeois, Friesekaas, Friesian, Friesla, Frinault, Fromage a Raclette, Fromage Corse, Fromage de Montagne de Savoie, Fromage Frais, Fruit Cream Cheese, Frying Cheese, Fynbo, Gabriel, Galette du Paludier, Galette Lyonnaise, Galloway Goat's Milk Gems, Gammelost, Gaperon a I'Ail, Garrotxa, Gastanberra, Geitost, Gippsland Blue, Gjetost, Gloucester, Golden Cross, Gorgonzola, Gornyaltajski, Gospel Green, Gouda, Goutu, Gowrie, Grabetto, Graddost, Grafton Village Cheddar, Grana, Grana Padano, Grand Vatel, Grataron d'Areches, Gratte-Paille, Graviera, Greuilh, Greve, Gris de Lille, Gruyere, Gubbeen, Guerbigny, Halloumi, Halloumy (Australian), Haloumi-Style Cheese, Harbourne Blue, Havarti, Heidi Gruyere, Hereford Hop, Herrgardsost, Herriot Farmhouse, Herve, Hipi Iti, Hubbardston Blue Cow, Humboldt Fog, Hushallsost, Iberico, Idaho Goatster, Idiazabal, II Boschetto al Tartufo, lie d'Yeu, Isle of Mull, Jarlsberg, Jermi Tortes, Jibneh Arabieh, Jindi Brie, Jubilee Blue, Juustoleipa, Kadchgall, Kaseri, Kashta, Kefalotyri, Kenafa, Kernhem, Kervella Affine, Kikorangi, King Island Cape Wickham Brie, King River Gold, Klosterkaese, Knockalara, Kugelkase, L'Aveyronnais, L'Ecir de I'Aubrac, La Taupiniere, La Vache Qul Rit, Laguiole, Lairobell, Lajta, Lanark Blue, Lancashire, Langres, Lappi, Laruns, Lavistown, Le Brin, Le Fium Orbo, Le Lacandou, Le Roule, Leafield, Lebbene, Leerdammer, Leicester, Leyden, Limburger, Lincolnshire Poacher, Lingot Saint Bousquet d'Orb, Liptauer, Little Rydings, Livarot, Llanboidy, Llanglofan Farmhouse, Loch Arthur Farmhouse, Loddiswell Avondale, Longhorn, Lou Palou, Lou Pevre, Lyonnais, Maasdam, Macconais, Mahoe Aged Gouda, Mahon, Malvern, Mamirolle, Manchego, Manouri, Manur, Marble Cheddar, Marbled Cheeses, Maredsous, Margotin, Maribo, Maroilles, Mascares, Mascarpone, Mascarpone (Australian), Mascarpone Torta, Matocq, Maytag Blue, Meira, Menallack Farmhouse, Menonita, Meredith Blue, Mesost, Metton (Cancoillotte), Meyer Vintage Gouda, Mihalic Peynir, Milleens, Mimolette, Mine-Gabhar, Mini Baby Bells, Mixte, Molbo, Monastery Cheeses, Mondseer, Mont D'or Lyonnais, Montasio, Monterey Jack, Monterey Jack Dry, Morbier, Morbier Cru de Montagne, Mothais a la Feuille, Mozzarella, Mozzarella (Australian), Mozzarella di Bufala, Mozzarella Fresh, in water, Mozzarella Rolls, Muenster, Munster, Murol, Mycella, Myzithra, Naboulsi, Nantais, Neufchatel, Neufchatel (Australian), Niolo, Nokkelost, Northumberland, Oaxaca, Olde York, Olivet au Foin, Olivet Bleu, Olivet Cendre, Orkney Extra Mature Cheddar, Orla, Oschtjepka, Ossau Fermier, Ossau-Iraty, Oszczypek, Oxford Blue, P'tit Berrichon, Palet de Babligny, Paneer, Panela, Pannerone, Pant ys Gawn, Parmesan (Parmigiano), Parmigiano Reggiano, Pas de I'Escalette, Passendale, Pasteurized Processed, Pate de Fromage, Patefine Fort, Pave d'Affinois, Pave d'Auge, Pave de Chirac, Pave du Berry, Pecorino, Pecorino in Walnut Leaves, Pecorino Romano, Peekskill Pyramid, Pelardon des Cevennes, Pelardon des Corbieres, Penamellera, Penbryn, Pencarreg, Pepper jack, Perail de Brebis, Petit Morin, Petit Pardou, Petit-Suisse, Picodon de Chevre, Picos de Europa, Pinconning, Piora, Pithtviers au Foin, Plateau de Herve, Plymouth Cheese, Podhalanski, Poivre d'Ane, Polkolbin, Pont I'Eveque, Port Nicholson, Port-Salut, Postel, Pouligny-Saint-Pierre, Pourly, Prastost, Pressato, Prince-Jean, Processed Cheddar, Provel, Provolone, Provolone (Australian), Pyengana Cheddar, Pyramide, Quark, Quark (Australian), Quartirolo Lombardo, Quatre-Vents, Quercy Petit, Queso Blanco, Queso Blanco con Frutas-Pina y Mango, Queso de Murcia, Queso del Montsec, Queso del Tietar, Queso Fresco, Queso Fresco (Adobera), Queso Iberico, Queso Jalapeno, Queso Majorero, Queso Media Luna, Queso Para Frier, Queso Quesadilla, Rabacal, Raclette, Ragusano, Raschera, Reblochon, Red Leicester, Regal de la Dombes, Reggianito, Remedou, Requeson, Richelieu, Ricotta, Ricotta (Australian), Ricotta Salata, Ridder, Rigotte, Rocamadour, Rollot, Romano, Romans Part Dieu, Roncal, Roquefort, Roule, Rouleau De Beaulieu, Royalp Tilsit, Rubens, Rustinu, Saaland Pfarr, Saanenkaese, Saga, Sage Derby, Sainte Maure, Saint-Marcellin, Saint-Nectaire, Saint-Paulin, Salers, Samso, San Simon, Sancerre, Sap Sago, Sardo, Sardo Egyptian, Sbrinz, Scamorza, Schabzieger, Schloss, Selles sur Cher, Selva, Serat, Seriously Strong Cheddar, Serra da Estrela, Sharpam, Shelburne Cheddar, Shropshire Blue, Siraz, Sirene, Smoked Gouda, Somerset Brie, Sonoma Jack, Sottocenare al Tartufo, Soumaintrain, Sourire Lozerien, Spenwood, Sraffordshire Organic, St. Agur Blue Cheese, Stilton, Stinking Bishop, String, Sussex Slipcote, Sveciaost, Swaledale, Sweet Style Swiss, Swiss, Syrian (Armenian String), Tala, Taleggio, Tamie, Tasmania Highland Chevre Log, Taupiniere, Teifi, Telemea, Testouri, Tete de Moine, Tetilla, Texas Goat Cheese, Tibet, Tillamook Cheddar, Tilsit, Timboon Brie, Toma, Tomme Brulee, Tomme d'Abondance, Tomme de Chevre, Tomme de Romans, Tomme de Savoie, Tomme des Chouans, Tommes, Torta del Casar, Toscanello, Touree de L'Aubier, Tourmalet, Trappe (Veritable), Trois Comes De Vendee, Tronchon, Trou du Cru, Truffe, Tupi, Turunmaa, Tymsboro, Tyn Grug, Tyning, Ubriaco, Ulloa, Vacherin-Fribourgeois, Valencay, Vasterbottenost, Venaco, Vendomois, Vieux Corse, Vignotte, Vulscombe, Waimata Farmhouse Blue, Washed Rind Cheese (Australian), Waterloo, Weichkaese, Wellington, Wensleydale, White Stilton, Whitestone Farmhouse, Wigmore, Woodside Cabecou, Xynotyro, Yarg Cornish, Yarra Valley Pyramid, Yorkshire Blue, Zamorano, Zanetti Grana Padano, and Zanetti Parmigiano Reggiano. In another aspect the cheese is selected from mozzarella, cheddar, cottage cheese, parmesan, and Swiss cheese. In another aspect the cheese is selected from semi-hard cheeses (such as continental cheeses and including gouda, edam, masdam), cheddar, cottage cheese, pasta filata cheese (such as mozzarella, pizza cheese), hard cheeses (such as parmesan, swiss type cheese, emmental type cheese), soft cheeses, Tvarog and lactic curd.

[0026] The milk used in the present process may be pasteurised or non-pasteurized (i.e. raw). In one aspect the milk is pasteurised milk. The milk may be standardised, homogenised or standardised and homogenised. It may also be the subject of one or more other treatments such ultra heat treatment (UHT).

[0027] The milk may be selected from, for example and without limitation, whole milk, reconstituted skim milk powder, skim milk, semi-skim milk and mixtures thereof.

[0028] The milk is obtained from any animal the milk of which is suitable for human consumption.

[0029] Such animals include, for example and without limitation, cow, camel, donkey, goat, horse, reindeer, sheep, water buffalo, and yak. The milk may also be a mixture of milks from one or more of the above-mentioned animals. In some aspects, the milk is selected from cow milk, sheep milk, goat milk and combinations thereof. In one aspect the milk is cow milk.

[0030] Acidification

[0031] It is a feature of the present invention that the milk is acidified with carbon dioxide, such that the pH of the acidified milk is about 6.2 to about 6.6. The milk is acidified by contacting the milk with a sufficient amount of carbon dioxide for time sufficient to acidify the milk to the required pH. The carbon dioxide may be delivered to and contact with the milk in any suitable manner. For example, the milk may be acidified by the addition of solid carbon dioxide to the milk. The solid carbon dioxide may be added in the form of pellets or larger blocks such as dry ice. In addition or in an alternative, the milk may be acidified by the bubbling of gaseous carbon dioxide through the milk. This may be performed by use of a sparging unit or any other appropriate apparatus. The operation and use of such units for contacting gaseous carbon dioxide with a liquid is well understood by one skilled in the art.

[0032] It is a requirement of the present invention that the pH of the milk is reduced from its initial value to the required range of 6.2 to 6.6. This will typically be achieved solely by addition of carbon dioxide to the milk i.e. the milk is acidified solely with carbon dioxide. In some aspects, carbon dioxide is the only source of acid used to acidify the milk. In certain aspects, the milk is acidified in part with carbon dioxide and in part with a secondary acidifying agent. Suitable secondary acidifying agent may be identified by one skilled in the art and include culture, such a lactic acid bacteria culture, organic or inorganic acids, such as hydrochloric acid or citric acid, and combinations thereof.

[0033] In respect of the acidification with carbon dioxide, one skilled in the art may calculate by routine experimentation by measurement of pH and without undue burden the amount of carbon dioxide required to acidify a given sample of milk to the desired extent. One skilled in the art may calculate by routine experimentation the amount of carbon dioxide and/or secondary acidifying agent needed to reach the desired measurement of pH and without undue burden the amount of carbon dioxide required to acidify a given sample of milk to the desired extent by routine methods that are well understood by a person of ordinary skill. In some aspects, a pH probe may be used to measure the pH of the milk and determine when the milk has reached the desired pH, at which time the addition of carbon dioxide and/or secondary acidifying agent can be stopped. In one aspect the present process further comprises the step of measuring the pH of the milk of step (i), and adding carbon dioxide to the milk in amount required to provide acidified milk having a pH of 6.2 to 6.6 when measured at a temperature of 29 to 38 C., wherein the amount of carbon dioxide added is 90 grams [0.2 lbs] of carbon dioxide per 1000 lbs of milk per 0.08 unit pH reduction.

[0034] In particular aspects the milk is acidified with carbon dioxide to a pH of, for example, 6.3 to 6.6, 6.35 to 6.6, pH of 6.3 to 6.55, pH of 6.4 to 6.6, pH of 6.35 to 6.57, pH of 6.4 to 6.57, pH of 6.41 to 6.6, pH of 6.41 to 6.57, pH of 6.41, 6.44 or 6.57.

[0035] In particular aspects the milk is acidified with carbon dioxide to a pH of, for example, 6.3 to 6.6, 6.35 to 6.6, pH of 6.3 to 6.55, pH of 6.4 to 6.6, pH of 6.35 to 6.57, pH of 6.4 to 6.57, pH of 6.41 to 6.6, pH of 6.41 to 6.57, pH of 6.41, 6.44 or 6.57, when measured at a temperature of 29 to 38 C.

[0036] The temperature at which the pH is measured may be from 31 to 36 C., such as 33 to 35 C., preferably about 34 C.

[0037] Coagulant

[0038] As discussed herein the inoculated acidified milk from which the cheese is prepared contains a coagulant. The coagulant comprises at least a microbial protease coagulating agent. The microbial protease coagulating agent may be the sole coagulant, in other words the coagulant consists of or consists essentially of the microbial protease coagulating agent. However, in one aspect the coagulant comprises the microbial protease coagulating agent and a secondary coagulating agent. The secondary coagulating agent is selected from animal-based coagulants such as rennet, vegetable-based coagulants such as Ficine and Bromeline, microbial based coagulants such as fermented produced chymosin coagulant or proteases obtained from e.g. Mucor miehei or Mucor pusillus or Cryphonectria parasitica.

[0039] The microbial protease coagulating agent is preferably a single enzyme coagulating agent. However, combinations of enzymes are also envisaged.

[0040] In one aspect, the microbial protease coagulating agent has k-casein cleaving activity. In one aspect the microbial protease coagulating agent predominantly has k-casein cleaving activity (that is the k-casein cleaving activity is greater than any side activities). In one aspect the microbial protease coagulating agent solely has k-casein cleaving activity (that is the microbial protease coagulating agent has no significant side activities). In one aspect the microbial protease coagulating agent is k-casein specific cleaving enzyme.

[0041] In one aspect, the microbial protease coagulating agent has pepsin activity. In one aspect the microbial protease coagulating agent predominantly has pepsin activity (that is the pepsin activity is greater than any side activities). In one aspect the microbial protease coagulating agent solely has pepsin activity (that is the microbial protease coagulating agent has no significant side activities). In one aspect the microbial protease coagulating agent is a pepsin.

[0042] In one aspect, the microbial protease coagulating agent has mucorpepsin (EC 3.4.23.23) activity. In one aspect the microbial protease coagulating agent predominantly has mucorpepsin (EC 3.4.23.23) activity (that is the mucorpepsin (EC 3.4.23.23) activity is greater than any side activities). In one aspect the microbial protease coagulating agent solely has mucorpepsin (EC 3.4.23.23) activity (that is the microbial protease coagulating agent has no significant side activities). In one aspect the microbial protease coagulating agent is a mucorpepsin (EC 3.4.23.23).

[0043] In yet other aspects the microbial protease coagulating agent is a single enzyme coagulating agent consisting of a mucorpepsin (EC 3.4.23.23) having k-casein specific cleaving activity. In yet other aspects the microbial protease coagulating agent consists essentially of a mucorpepsin (EC 3.4.23.23) having k-casein specific cleaving activity. In further aspects the microbial protease coagulating agent is at least mucorpepsin (EC 3.4.23.23), such as MARZYME available from Danisco A/S. Preferably the microbial protease coagulating agent consists of mucorpepsin (EC 3.4.23.23), such as MARZYME available from Danisco A/S.

EXAMPLES

[0044] The invention will now be described with reference to the following non-limiting example.

[0045] The history regarding conversion from bulk starter to DVI (Direct Vat Inoculation) culture system has shown that generally the pH acidification curve is slower from the beginning when converting to DVI culture systems from bulk starter and can result in higher end pHs and higher moistures (sometimes out of specification) because of this in-process condition. In addition, we have seen in commercial applications that the in-process whey fats can be increased 0.3% average (from 0.5% to 0.8%) when using DVI vs. bulk starter resulting mainly from this increase in pH from the beginning of the process, hindering coagulation efficiency. This has a direct impact on further processing of the whey (fat needs to be removed prior to further processing and higher whey fats can slow down this process) as well as a financial impact to the processing plant by losing this component (fat) in the cheese.

[0046] This invention has employed the use of carbon dioxide when using DVI culture system to create carbonic acid to lower the starting pH and thus lower the in-process whey fats and mimic the pH acidification curves and final pH and moistures in the cheese. The initial testing was done by lowering the milk pH to 6.55 target using dry ice (solid carbon dioxide) during the milk fill step in the process. This is a 0.08 average reduction in the starting milk pH. Initial work was done on Monterey Jack and White and Colored Cheddar cheeses in a commercial setting.

[0047] The initial testing was done using 55,000 pound capacity horizontal agitated enclosed cheese vats. Milk is standardized with a protein concentrate obtained from ultrafiltration technology to a milk fat target of 5.0%, protein target of 4.45%, lactose target of 4.6%, and pH target of 6.63. Depending upon casein:fat ratio, total solids can range from 14.8-15.5%, thus milk equivalent in the vat is 68,000-75,000 pounds. Standardized milk is pasteurized at 165 F. for minimum of 16 seconds and cooled to vat set temperature of 88-90 F. and pumped into vat. Dry ice is added when vat fill reaches 6000-8000 lbs. Dry ice is added either via pellet form or chunks of 33 pieces and allowed to melt into the vat milk. Dry Ice is added at a rate of 15 lbs per 72,000 pounds milk equivalent. Color (if needed) and calcium chloride are added to the vat as well as frozen pellet DVI cultures which consist of either mesophilic strains or a mesophilic/thermophilic strain blend of frozen pellets. Usage rate is 2375 DCU (Danisco Culture Unitsa commercial unit) per 75,000 pounds milk equivalent. Once the vat is full, the milk pH is measured. Diluted coagulant (FPCFermentation Produced Chymosinor Marzyme Supreme microbial) is added via automated system to the vat and stirred in for 3-4 minutes, reversing agitators for 45 seconds to promote slowing of the milk mass, to achieve more even coagulation. Vat is allowed to coagulate and is cut at proper time when a spatula has made a clean slice of the cheese mass. Cheese mass is cut into cubes and allowed to heal for 1-5 minutes. Jacketed heat is applied to the vat and the cheese and whey temperature is brought up to 101-103 F. Once cooking is complete, the cheese and whey is transferred to an enclosed belt system to form a mat of cheese and drain the whey. The pump over pH is taken at this time as well as pump over pH of the curd and whey. In addition, the whey fat is measured. In the case of Monterey Jack (MJ), a warm water spray (86-88 F.) is put on the curd in this vessel to allow for reduction of the concentration of lactose to control moisture and pH of the final product. A sufficient amount of time is employed to mat the curd and drain the whey, then the cheese is automatically cut using a mill machine into approximately -14 pieces of cheese curd. CM whey fat (cheese machine whey fat, also called whey fat before salt) is taken at this time as well as mill pH of curd and whey. These milled cheese pieces are transferred to a salting belt which applies 2 salting applications via automated system and then allows approximately 10 minutes of mellowing time. Salted curd is then transferred to a distributor which transfers the curds to towers which press the cheese curd together and vacuum out the whey to produce 40 lbs blocks of pressed cheese. These 40 lbs blocks are bagged, weighed, labelled and then sent to a pre-cooler which brings the temperature to near 50 F. Packaged blocks go from the pre-cooler to be palletized and onto racks in a 35 degree Fahrenheit cooler warehouse until shipment. Finished goods samples are run for moisture, fat, salt and pH at 5 days after make.

[0048] Results of the initial testing showed that by using dry ice (carbon dioxide) to lower the initial vat milk pH, the DVI cultures can more mimic the bulk starter culture system make resulting in final moistures and pHs which meet specification. In addition, the whey fats results when using DVI and employing the carbon dioxide technology can reduce the whey fats to levels seen with bulk starter. Further, Marzyme Supreme can have similar results as FPC when looking at whey fat retention in the curd and employing the carbon dioxide technology.

[0049] The following results were seen using carbon dioxide technology with DVI vs. bulk starter and Marzyme Supreme vs. FPC in commercial vats using the procedures outlined above:

TABLE-US-00001 Total number Milk Whey CM Whey Day Type DVI/BS of Vats pH Fat Fat 1 MJ CO2 + DVI + FPC 5 6.538 0.184 0.46 1 MJ No CO2 + Bulk 28 6.628 0.174 0.516 Starter + FPC 2 WC CO2 + DVI + 5 6.57 0.22 0.50 Marzyme Supreme 2 WC No CO2 + Bulk 20 6.643 0.196 0.514 Starter + Marzyme Supreme 4 CC CO2 + DVI + FPC 36 6.59 0.208 0.44 5 CC CO2 + Bulk 35 6.58 0.18 0.387 Starter + FPC [0050] Where MJ=Monterey Jack; WC=White Cheddar; CC=Colored Cheddar [0051] CO.sub.2 is 15 lbs dry ice per 75,000 pounds milk equivalent [0052] Where Whey Fat is measured during pump over of curd and whey to enclosed belt system, and CM Whey Fat is taken at milling of the cheese curd [0053] Whey Fat and CM Whey Fat are expressed in % fat by weight values of the whey

[0054] Since final moistures and pHs are important for specification to the cheese processing plant, averages for days 4 and 5 were collected and analyzed for vats run with DVI vs. bulk starter. Specification for moisture cannot go above 39% for standard of identity of cheddar cheese, but of course there is an advantage to reach as near 39% because of final payment of cheese (price per pound final product). In addition, standard of identity for cheddar cheese for pH is below 5.35 pH. Commercial dairies target final pH between 5.05 and 5.20 for mild cheddar cheese. Final analysis is reflected below:

TABLE-US-00002 Total Final number Moisture Final Day Type Vat treatment of Vats in % pH 4 CC CO2 + DVI + FPC 36 37.64 5.131 5 CC CO2 + Bulk Starter + FPC 35 37.72 5.094

[0055] Both DVI and Bulk Starter using the Carbon Dioxide technology were within specification and within the normal parameters of the commercial operations of this facility.

[0056] As benefits were seen with this addition of Carbon Dioxide with DVI culture systems and Marzyme Supreme, additional trials have been completed by dropping the starting milk pH to 6.40-6.45 pH. Findings have been a drop by over 50% in the amount of Marzyme Supreme needed for coagulation (75 weight oz. per 75,000 pound milk equivalents per vat drop to 35 weight oz. per 75,000 milk equivalents per vat); elimination of calcium chloride; decrease in CM whey fats; final moisture and pHs meeting specification targets; fines reduction of 25-30%, increasing final cheese yield.

[0057] Carbon dioxide was added via sparging unit to these test vats at a rate of 17 lbs per vat at about 50 pounds per square inch pressure. Milk equivalent to the vat was 74,500 pounds.

[0058] The following data is from days 9 and 10 reflecting these changes:

TABLE-US-00003 Total number Milk Whey CM Whey Day Type Milk additions of Vats pH Fat Fat 9 MJ CO2 + DVI + 13 6.411 0.259 0.442 Marzyme Supreme 10 WC CO2 + DVI + 10 6.442 0.312 0.411 Marzyme Supreme [0059] Whey Fat and CM Whey Fat are expressed in % fat by weight values of the whey

[0060] Additional make information is as follows:

TABLE-US-00004 Total number PO Mill Final Final Day Type Milk additions of Vats pH pH pH Moisture 9 MJ CO2 + DVI + 13 Recorded 6.109 5.538 5.189 42.17% Marzyme Supreme Target 6.1-.25 5.5-.6 5.15-.25 41-43% 10 WC CO2 + DVI + 10 Recorded 6.225 5.385 5.113 36.87% Marzyme Supreme Target 6.1-.25 5.4-.6 5.05-.20 36.5-39%.sup.

[0061] All make parameters fall within commercial production targets, with production on day 10 the mill pH a little low, however final pH and moisture are within targets.

[0062] This invention employs the fact that by adding carbon dioxide to vat milk we can reduce the milk pH by conversion to carbonic acid in the system. Using Dry Ice on an experimental design can mimic usage and in-process parameters such as with a sparging unit. By addition of carbon dioxide, the DVI method of culture addition can mimic bulk starter for in-process make parameters and final pH and moisture specification. Marzyme Supreme can replace FPC as coagulant by reducing in-process whey fats, thus leaving more fat in the cheese as well as reducing the usage rate substantially.

[0063] Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in chemistry, biology or related fields are intended to be within the scope of the following claims.