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Tepzz¥5 ¥664A T (19) TZZ¥_¥_T (11) EP 3 513 664 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 24.07.2019 Bulletin 2019/30 A23L 29/206 (2016.01) A23L 25/00 (2016.01) A23C 19/055 (2006.01) (21) Application number: 18203176.5 (22) Date of filing: 13.01.2014 (84) Designated Contracting States: • CASINO, Monte AL AT BE BG CH CY CZ DE DK EE ES FI FR GB San Bruno, CA California 94066 (US) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO • BROWN, Patrick O’Reilly PL PT RO RS SE SI SK SM TR Stanford, CA California 94305 (US) • EISEN, Michael (30) Priority: 11.01.2013 US 201361751818 P Berkeley, CA California 94703 (US) • COHN, Elysia (62) Document number(s) of the earlier application(s) in Mountain View, CA California 94041 (US) accordance with Art. 76 EPC: • PREVOT, Jean 14738061.2 / 2 943 077 Foster City, CA California 94404 (US) (71) Applicant: Impossible Foods Inc. (74) Representative: Novitas Patent AB Redwood City, CA 94063 (US) P.O. Box 55557 102 04 Stockholm (SE) (72) Inventors: • HOLZ-SCHIETINGER, Celeste Remarks: East Palo Alto, CA California 94303 (US) •This application was filed on 29-10-2018 as a • KLAPHOLZ, Sue divisional application to the application mentioned Stanford, CA California 94305 (US) under INID code 62. • VARADAN, Ranjani •Claims filed after the date of filing /date of receipt of Fremont, CA California 94536 (US) the divisional application (Rule 68(4) EPC). (54) METHOD OF PRODUCING A FLAVOURED CULTURED NON-DAIRY PRODUCT (57) Provided herein are methods and compositions for the production of cheese replicas. Generally the cheese replicas are produced by inducing the enzymatic curdling of non-dairy milks. EP 3 513 664 A1 Printed by Jouve, 75001 PARIS (FR) EP 3 513 664 A1 Description CROSS-REFERENCE TO RELATED APPLICATIONS 5 [0001] This application claims priority to U.S. Application Serial No. 61/751,818, filed January 11, 2013, and is related to co-pending Application Serial No PCT/US12/46552, filed July 12, 2012, all of which are incorporated herein by refer- ence. BACKGROUND OF THE INVENTION 10 [0002] Cheese making has relied on dairy milks as the major ingredient for more than 4000 years. Dairy cheese is usually made from curds formed from dairy milk. Dairy milks can readily be made to form curds suitable for making cheese by contacting the dairy milk with rennet (an aspartic protease which cleaves kappa-casein) at mildly acidic pH. Some cheeses, e.g., cream cheese, cottage cheese and paneer, are made without rennet. In the absence of rennet, 15 dairy cheese may be induced to curdle with acid (e.g., lemon juice, vinegar, etc.) or a combination of heat and acid. Acid coagulation can also occur naturally from starter culture fermentation. The strength of the curds depends on the type of coagulation. Most commercially produced cheeses use some type of rennet (animal, vegetable or microbial-derived) in their production. Commodity cheeses or "processed cheeses" such as bulk cheddar, food-service mozzarella pizza and "cheese products," or "cheese foods" such as American cheese, American singles, Velveeta, and Cheese Whiz are 20 typically produced from dairy-derived ingredients and other additives using industrial processes which sometimes little resemble traditional cheese making. [0003] The global dairy sector contributes an estimated 4 percent to the total global anthropogenic green house gas emissions. Producing 1 kg of cheddar cheese requires an average of 10,000 L of fresh water. Additionally, many indi- viduals cannot digest and metabolize lactose. In these individuals enteric bacteria ferment the lactose, resulting in various 25 abdominal symptoms, which can include abdominal pain, bloating, flatulence, diarrhea, nausea, and acid reflux. Addi- tionally, the presence of lactose and its fermentation products raises the osmotic pressure of the colon contents. Ap- proximately 3.4% of children in the U.S.A. are reported to have allergies to dairy milks. Many individuals choose to avoid milk for ethical or religious reasons. [0004] Non-dairy milks, including plant-derived milks avoid many of the environmental, food sensitivity, ethical and 30 religious problems associated with dairy milk and they can be made free of lactose, making the generation of dairy substitutes using the plant derived milks attractive. However, rennet is not an effective agent for inducing non-dairy proteins or emulsions, including plant-derived milks (e.g., almond milk, chestnut milk, pecan milk, hazelnut milk, cashew milk, pine nut milk, or walnut milk), to curdle. Consequently, traditional cheese making techniques have not been suc- cessfully used to produce non-dairy cheese replicas. 35 [0005] Flavor and aroma in dairy cheese results in part from the degradation of lactose, proteins and fats, carried out by ripening agents, which include bacteria and enzymes in the milk, bacterial cultures added during the cheese-making process, rennet, other proteases, lipases, added molds and/or yeasts and bacteria and fungi that opportunistically colonize the cheese during ripening and aging. In addition, the bacterial cultures and fungi used in traditional dairy cheesemaking use microorganisms adapted to growing and producing flavors in dairy milks. Hence, traditional cheese 40 culturing techniques have not been successfully used to produce non-dairy cheese replicas. [0006] Cheese replicas made principally of non-dairy ingredients are commercially available. Many of these cheese replicas include some dairy ingredients, for example, casein. Some commercially available cheese replicas contain no animal products. These include fermented cheese replicas made from nut milks from which insoluble carbohydrates have not been effectively removed, and made without using a protein crosslinking agent and several products in which 45 a starch is a principal ingredient or containing agar, carrageenan or tofu to provide the desired texture. The few fermented products contain Lactobacillus acidophilus, a microbe often used in dairy yogurts. Most tasters consider none of the currently available cheese replicas to adequately replicate the taste, aroma and mouthfeel of dairy cheeses. [0007] Complex carbohydrates in currently available cheese replicas made from nut milks have unfavorable effects on the texture, resulting in a product with a grainy mouthfeel and lacking the creaminess of dairy cheeses. 50 [0008] Starches that comprise the major gelling agent in many currently available cheese replicas lead to a relatively high carbohydrate content, which may be undesirable to consumers, for example those wishing to limit carbohydrate intake. [0009] Because of these deficiencies, there is currently no cheese replica that is acceptable to most consumers as an alternative to traditional dairy cheeses. 55 [0010] Thus, it is clear that there is a great need in the art for an improved method and system for producing non-dairy cheese replicas while avoiding the shortcomings and drawbacks of the cheese replicas that have previously been available to consumers. 2 EP 3 513 664 A1 SUMMARY OF THE INVENTION [0011] The invention relates to methods and compositions for producing non-dairy milk and cheese products, including without limitation, plant-derived milk and cheese products, as an alternative to dairy products for human consumption. 5 [0012] In one aspect, this document features a non-dairy cheese replica that includes a coacervate comprising one or more isolated and purified proteins from a non-animal source. The one or more isolated and purified proteins can be plant proteins (e.g., seed storage proteins, pea proteins, Lupine proteins, proteins from a legume, chickpea proteins, or lentil proteins. The pea proteins can include pea vicilins and/or pea legumins. The non-dairy cheese replica can include one or more microbes selected from bacteria, molds, and yeast. 10 [0013] This document also features a non-dairy cheese replica that includes a cold set gel comprising one or more isolated and purified proteins from a non-animal source and a salt. The non-dairy cheese replica can include one or more heat-labile ingredients such as one or more fats, microbes, volatile compounds, or enzymes. The non-dairy cheese replica can include one or more microbes selected from bacteria, molds, and yeast. [0014] The one or more microbes in the non-dairy cheese replicas can be selected from the group consisting of a 15 Penicillium species, a Debaryomyces species, a Geotrichum species, a Corynebacterium species, a Streptococcus species, a Verticillium species., a Kluyveromyces species, a Saccharomyces species, a Candida species, a Rhodospori- dum species, a Cornybacteria species, a Micrococcus species, a Lactobacillus species, a Lactococcus species, a Sta- phylococcus species, a Halomonas species, a Brevibacterium species, a Psychrobacter species, a Leuconostocaceae species, a Pediococcus species, a Propionibacterium species, and a lactic acid bacterium. 20 [0015] In some embodiments, the one or more microbes are selected from the group consisting of Lactococcus lactis lactis (LLL), Leuconostoc mesenteroides cremoris (LM), Lactococcus lactis cremoris (LLC), Pediococcus pentosaceus, Clostridium butyricum, Lactobacillus delbrueckii lactis, Lactobacillus delbrueckii bulgaricus, Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus rhamnosus, Staphylococcus xylosus (SX), Lactococcus lactis biovar diacetylactis (LLBD), Penicillium roqueforti, Penicillium candidum, Penicillium camemberti, Penicillium nalgiov- 25 ensis Debaryomyces hansenii, Geotrichum
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