Milk product

Abstract

The present invention relates to bio-transformed milk products, for example evaporated milk or similar products, that have improved properties and / or characteristics relative to equivalent non-bio-transformed milk products; or have comparable sensory characteristics to non-bio- transformed milk products that comprise greater quantities of milk solids. The invention also encompasses methods for making said bio-transformed milk products.

Description

[0001] MILK PRODUCT

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to bio-transformed milk products that have improved properties and / or characteristics relative to equivalent non-bio-transformed milk products; or have comparable sensory characteristics to non-bio-transformed milk products that comprise greater quantities of milk solids. The invention also encompasses methods for making said bio-transformed milk products.

[0004] BACKGROUND OF THE INVENTION

[0005] Milk products, in particular concentrated milk products, such as evaporated milk, condensed milk, and creamers are manufactured and consumed globally.

[0006] The ability to produce a product that has desirable consumer characteristics, such as longevity, mouthfeel and creaminess, is of key importance.

[0007] The sensory characteristics of milk products are tightly associated with components of the product. Typically, products with greater quantities of milk solids, such as milk protein and milk fats - or other non-milk solids - are associated with increased perception of creaminess and / or an improved mouthfeel, which is desirable for numerous applications.

[0008] In concentrated milk products, the content of milk solids (e.g., proteins and / or fats) is increased relative to those found in fresh / unprocessed milk. As a direct consequence of this, these products are typically associated with desirable sensory characteristics such as creaminess, which facilitate their use in specific applications and appeal to consumers.

[0009] Whilst increasing the milk solids content in concentrated milk products yields favourable sensory characteristics, it may be more costly as greater quantities of raw materials (e.g., milk) are required; and can lead to a reduction in the stability, such as the long term storage stability, of the product, or the ability of the product to withstand heat treatment (e.g., sterilization).

[0010] There is, as such, an unmet need for concentrated milk products that have a reduced economic cost, and / or have improved stability whilst retaining consistent and favourable sensory profiles that consumers demand. Moreover, there is an unmet need for methods / processes for making such products. SUMMARY OF THE INVENTION

[0011] The present inventors have surprisingly found that treatment of concentrated milk products and their precursors with a transglutaminase enzyme results in concentrated milk products with improved sensory and stability characteristics. More specifically, bio-transformed (i.e., transglutaminase treated) milk products having lower total milk solids and / or lower total milk protein content retained the sensory characteristics of non-bio-transformed milk products with a higher total milk solids and / or lower total milk protein content, thereby producing a consistent consumer product that requires less milk-derived solids to manufacture.

[0012] In other words, the inventors have surprisingly found that treatment of concentrated milk product precursors allows the production of concentrated milk products that require lower amounts of milk solids, such as milk proteins. These improved products retain desirable consumer characteristics whilst simultaneously facilitating a reduction in economic costs associated with reducing raw material (i.e., milk solids) requirements.

[0013] The present invention may therefore provide advantages associated with milk products that are cheaper to produce, have improved stability, and / or retain a favourable sensory profile.

[0014] In one aspect, there is provided a method for the production of a concentrated milk product having:

[0015] (a) a milk protein content of at least 3.5% by weight; and / or

[0016] (b) a total milk solids content of at least 15% by weight; wherein the method comprises the step of adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product.

[0017] In one aspect, there is provided a concentrated milk product produced by a method of the present invention.

[0018] In a further aspect, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0019] (a) a milk protein content of at least 3.5% by weight; and / or

[0020] (b) a total milk solids content of at least 15% by weight; and

[0021] (c) cross-linked milk proteins; and

[0022] (d) trace amounts of an inactivated or non-functional transglutaminase; and optionally

[0023] (e) carrageenan. By providing the concentrated milk products and the methods for their manufacture herein, the present invention addresses and overcomes the unmet need for concentrated milk products that retain favourable sensory profiles whilst being more commercially economical to manufacture than currently available products.

[0024] BRIEF DESCRIPTION OF THE FIGURES

[0025] Figure 1 - Illustrative manufacturing flow diagram

[0026] An illustrative manufacturing flow diagram demonstrating a typical process for the manufacture of concentrated milk products. * indicates that this step includes homogenization, wet-mixing, standardization, and other processes; **indicates that after UHT sterilization, an aseptic homogenization step could optionally be used.

[0027] Figure 2 - Bio-transformation facilitates a reduction in milk solids

[0028] Figure 2 illustrates an example work flow for the preparation of bio-transformed concentrated milk products such as those tested in Example 2.

[0029] Figure 3 - Reduced sedimentation in bio-transformed milk products

[0030] Reduced sedimentation of bio-transformed milk products during storage. Time and temperature dependent sedimentation of concentrated milk products was assessed by visual inspection. Non-bio-transformed (‘sample T) concentrated milk products succumb to sedimentation overtime, whereas bio-transformed (‘sample 2’) concentrated milk products are more stable and do not produce sediment.

[0031] Figure 4 - Increasing pH increases heat stability

[0032] Figure 4 illustrates an example work flow for the preparation of bio-transformed concentrated milk products, such as those tested in Example 4, which have a variety of different pH values prior to sterilization.

[0033] DETAILED DESCRIPTION OF THE INVENTION

[0034] It is an object of the present invention to provide concentrated milk products that retain favourable sensory profiles; are economical to manufacture; and have adequate, or improved, stability by providing such products and methods for their manufacture.

[0035] Concentrated milk products

[0036] The present invention is concerned with concentrated milk products and methods for their manufacture. Herein, a concentrated milk product may refer to any product that is derived from milk and has undergone at least one process whereby one or more substituents of milk has been concentrated (i.e. , increased) relative to unprocessed fresh milk.

[0037] In particular, concentrated milk products of the present invention may refer to milk products wherein the total amount of milk solids and / or milk proteins is increased relative to fresh or unprocessed milk, resulting in altered sensory characteristics (e.g., increased creaminess).

[0038] Well-known concentrated milk products include condensed milks, evaporated milks, and creamers.

[0039] In one embodiment, the concentrated milk product is a condensed milk product.

[0040] In one embodiment, the concentrated milk product is an evaporated milk product.

[0041] In one embodiment, the concentrated milk product is a creamer product.

[0042] The definitions and / or characterization of specific milk products may vary between geographical regions and under different regulatory authorities, however the person skilled in the art is nonetheless able to determine what constitutes a specific concentrated milk product based on the components thereof (e.g., milk protein content, etc.).

[0043] Suitably, the concentrated milk product may have a milk protein content of at least 3.5% by weight.

[0044] Suitably, the concentrated milk product may have a milk protein content of at least 4% by weight.

[0045] Suitably, the concentrated milk product may have a milk protein content of at least 4.5% by weight.

[0046] Suitably, the concentrated milk product may have a milk protein content of at least 5% by weight.

[0047] Suitably, the concentrated milk product may have a milk protein content of at least 3% by weight, such as at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10% by weight. Suitably, the concentrated milk product may have a milk protein content of about 3% by weight to about 10% by weight, such as about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 9%, about 6% to about 8%, or about 6% to about 7% by weight.

[0048] Suitably, the concentrated milk product may have a total milk solids content of at least 15% by weight.

[0049] Suitably, the concentrated milk product may have a total milk solids content of at least 16% by weight.

[0050] Suitably, the concentrated milk product may have a total milk solids content of at least 17% by weight.

[0051] Suitably, the concentrated milk product may have a total milk solids content of at least 18% by weight.

[0052] Suitably, the concentrated milk product may have a total milk solids content of at least 19% by weight.

[0053] Suitably, the concentrated milk product may have a total milk solids content of at least 19.5% by weight.

[0054] Suitably, the concentrated milk product may have a total milk solids content of at least 20% by weight.

[0055] Suitably, the concentrated milk product may have a total milk solids content of at least 20% by weight.

[0056] Suitably, the concentrated milk product may have a total milk solids content of at least 20.5% by weight.

[0057] Suitably, the concentrated milk product may have a total milk solids content of about 20.5% by weight. Suitably, the concentrated milk product may have a total milk solids content of at least 21% by weight.

[0058] Suitably, the concentrated milk product may have a total milk solids content of about 15% by weight, about 15.5% by weight, about 16% by weight, 16.5% by weight, about 17% by weight, about 17.5% by weight, about 18% by weight, about 18.5% by weight, about 19% by weight, about 19.5% by weight, about 20% by weight, about 20.5% by weight, about 21% by weight, about 21.5% by weight, about 22% by weight.

[0059] Suitably, the concentrated milk product may have a total milk solids content of about 15% by weight to about 30% by weight, such as about 15% to about 25%, about 15% to about 24%, about 15% to about 23%, about 15% to about 22%, about 15% to about 21 %, about 15% to about 20%, about 17% to about 30%, about 17% to about 25%, about 17% to about 24%, about 17% to about 23%, about 17% to about 22%, about 17% to about 21 %, about 17% to about 20%, about 19% to about 30%, about 19% to about 25%, about 19% to about 24%, about 19% to about 23%, about 19% to about 22%, about 19% to about 21 %, about 19% to about 20%, about 20% to about 30%, about 20% to about 25%, about 20% to about 24%, about 20% to about 23%, about 20% to about 22%, or about 20% to about 21%.

[0060] Suitably, the concentrated milk product may have a total milk solids content of about 19% by weight to about 22% by weight.

[0061] Suitably, the concentrated milk product may have a total milk solids content of about 19% by weight to about 21 % by weight.

[0062] Suitably, the concentrated milk product may have a total milk solids content of about 20% by weight to about 21 % by weight.

[0063] In one embodiment there is provided a concentrated milk product having:

[0064] (a) a milk protein content of at about 3% by weight to about 10% by weight, such as about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 9%, about 6% to about 8%, or about 6% to about 7% by weight; and / or (b) a total milk solids content of at about 15% by weight to about 30% by weight, such as about 15% to about 25%, about 15% to about 24%, about 15% to about 23%, about 15% to about 22%, about 15% to about 21 %, about 15% to about 20%, about 17% to about 30%, about 17% to about 25%, about 17% to about 24%, about 17% to about 23%, about 17% to about 22%, about 17% to about 21%, about 17% to about 20%, about 19% to about 30%, about 19% to about 25%, about 19% to about 24%, about 19% to about 23%, about 19% to about 22%, about 19% to about 21%, about 19% to about 20%, about 20% to about 30%, about 20% to about 25%, about 20% to about 24%, about 20% to about 23%, about 20% to about 22%, or about 20% to about 21%.

[0065] In one embodiment, the concentrated milk product has a total lipid content of about 1% to about 10% by weight, such as about 1 % to about 9%, about 1 % to about 8%, about 1% to about 7%, about 1 % to about 6%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, 5% to about 9%, about 5% to about 8%, about 5% to about 7%, or about 5% to about 6%.

[0066] In one embodiment, the concentrated milk product has a total milk lipid and / or fat content of about 1 % to about 10% by weight, such as about 1 % to about 9%, about 1% to about 8%, about 1% to about 7%, about 1% to about 6%, about 1 % to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, 5% to about 9%, about 5% to about 8%, about 5% to about 7%, or about 5% to about 6%.

[0067] In one embodiment, the concentrated milk product has a total lipid content of about 4% to about 5% by weight.

[0068] In one embodiment, there is provided a concentrated milk product produced by a method according to the present invention. The methods of the invention concern bio-transformation of milk proteins as a result of transglutaminase activity. Hence, a concentrated milk product or any post-transglutaminase treatment precursor thereof may be distinguished by the presence of inter- and intramolecular cross-links between any one or more milk proteins / polypeptides. As discussed further herein, the presence of such cross-links could be identified by the person skilled in the art in numerous ways.

[0069] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0070] (a) a milk protein content of at least 3.5% by weight; and / or

[0071] (b) a total milk solids content of about 15% by weight to about 26% by weight; and,

[0072] (c) cross-linked milk proteins; and

[0073] (d) trace amounts of an inactivated or non-functional transglutaminase.

[0074] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0075] (a) a milk protein content of at least 3.5% by weight; and / or

[0076] (b) a total milk solids content of about 15% by weight to about 26% by weight; and,

[0077] (c) cross-linked milk proteins.

[0078] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0079] (a) a milk protein content of at about 3% by weight to about 10% by weight, such as about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 9%, about 6% to about 8%, or about 6% to about 7% by weight; and / or

[0080] (b) a total milk solids content of at about 15% by weight to about 30% by weight, such as about 15% to about 25%, about 15% to about 24%, about 15% to about 23%, about 15% to about 22%, about 15% to about 21 %, about 15% to about 20%, about 17% to about 30%, about 17% to about 25%, about 17% to about 24%, about 17% to about 23%, about 17% to about 22%, about 17% to about 21%, about 17% to about 20%, about 19% to about 30%, about 19% to about 25%, about 19% to about 24%, about 19% to about 23%, about 19% to about 22%, about 19% to about 21%, about 19% to about 20%, about 20% to about 30%, about 20% to about 25%, about 20% to about 24%, about 20% to about 23%, about 20% to about 22%, or about 20% to about 21%; and,

[0081] (c) cross-linked milk proteins.

[0082] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0083] (a) a milk protein content of at about 3% by weight to about 10% by weight; and / or

[0084] (b) a total milk solids content of at about 15% by weight to about 30% by weight; and,

[0085] (c) cross-linked milk proteins.

[0086] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has:

[0087] (a) a milk protein content of at about about 6% to about 7% by weight; and / or

[0088] (b) a total milk solids content of at about 20% to about 21%; and,

[0089] (c) cross-linked milk proteins.

[0090] In one embodiment, the concentrated milk product has about 20.5% total milk solids, about 5.2% milk fat, and cross-linked milk proteins.

[0091] In one embodiment, the concentrated milk product comprises trace amounts of an inactivated or non-functional transglutaminase.

[0092] It will be understood that the transglutaminase may be inactivated or otherwise rendered non enzymatically functional in a method according to the invention (e.g., heat treatment) such that it can no longer act to catalyse transglutamination. Residual inactivated transglutaminase protein / peptides may be detectable in the concentrated milk product using highly sensitive detection techniques such as western blotting or mass spectrometry.

[0093] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has a total milk solids content of about 20% by weight to about 21 % by weight, and comprises cross-linked milk proteins.

[0094] In one embodiment, there is provided a concentrated milk product, wherein the concentrated milk product has a total milk solids content of about 20% by weight to about 21 % by weight, and comprises trace amounts of inactivated or non-functional transglutaminase. The concentrated milk products of the invention may be processed to remove one or more components present in milk, e.g., by skimming to reduce fat or any other processes.

[0095] Certain products, such as creamers, may be processed to remove lactose or other components present in milk.

[0096] In one embodiment, the concentrated milk product has been processed to remove one or more components present in fresh milk.

[0097] In one embodiment, the concentrated milk product is lactose free or substantially lactose free.

[0098] In one embodiment, the concentrated milk product is not a fermented milk product.

[0099] In one embodiment, the concentrated milk product is not derived from any fermented milk product precursors.

[0100] In one embodiment, the concentrated milk product is not a cheese product or a yoghurt product. In one embodiment, the concentrated milk product is not a cheese product In one embodiment, the concentrated milk product is not a yoghurt product.

[0101] Additional non-milk components

[0102] The concentrated milk product may comprise any number of suitable components that are not directly derived from milk.

[0103] Many suitable food-grade additives are known to the person skilled in the art.

[0104] In one embodiment, the concentrated milk product comprises carrageenan.

[0105] Stabilizers and or buffers may be added to the concentrated milk product.

[0106] In one embodiment the concentrated milk product comprises phosphates, such as sodium or potassium phosphates.

[0107] In one embodiment the concentrated milk product comprises sodium phosphate, potassium phosphate, or a mixture of sodium and potassium phosphates. Phosphates may be added to the milk product in order to improve stability. However, the addition of phosphates in high quantities can result in a negative consumer perception of increased saltiness.

[0108] The present inventors surprisingly found that the bio-transformed concentrated milk products of the present invention require reduced amounts of phosphates to provide heat stability and / or long term storage stability.

[0109] Suitable amounts of stabilizers may be determined by a person skilled in the art. Further, it will be understood that stability may be assessed with respect to different parameters (e.g., heat stability [see e.g., Example 4 herein] or temporal stability [see e.g., Examples 2 and 3 herein]). As such, different types of stability may be independently assessed and / or determined.

[0110] In one embodiment the concentrated milk product comprises 0.01 % to 0.3% by weight phosphates.

[0111] In one embodiment the concentrated milk product comprises 0.02% to 0.2% phosphates.

[0112] In one embodiment the concentrated milk product comprises less than 0.16% phosphates.

[0113] In one embodiment the concentrated milk product has a pH of 6.1 or above.

[0114] The pH of the concentrated milk product may be different to that of a precursor. For example, processes such as sterilization may change the pH of a precursor that was at a different pH or had been adjusted to a specific pH.

[0115] Characteristics of the concentrated milk products

[0116] The concentrated milk products of the present invention have numerous advantageous characteristics.

[0117] Firstly, the bio-transformed concentrated milk products have similar sensory profiles to non- bio-transformed milk products that comprise similar amounts of milk solids and / or milk proteins. As such, the concentrated milk products of the invention are able to retain a sensory profile that is desired by consumers whilst utilizing less raw materials. Secondly, the bio-transformed concentrated milk products of the invention have improved stability, which, for example, enables their long term storage and results in an improved shelf life.

[0118] The improved stability of the bio-transformed concentrated milk products of the invention is apparent in at least two different ways: firstly, heat stability of the product (or bio-transformed precursors thereof) is increased such that they can withstand heat treatment (e.g., sterilization) that may be required; secondly, the temporal stability of the product (or bio-transformed precursors thereof) is increased such that they can tolerate long term storage without a reduction or change in the product characteristics.

[0119] In particular, long term storage stability may be assessed on the basis of:

[0120] • Unaltered sensory profile (e.g., taste, mouthfeel, creaminess, smell) as compared with a newly prepared product;

[0121] • Absence of liquid phase separation;

[0122] • Absence of the formation of solid phase separation (e.g., sediment formation); and / or

[0123] • Unaltered appearance (e.g., colour) as compared with a newly prepared product.

[0124] It will be understood that long term stability may be assessed on the basis of the absence of essentially any change of a product over time as compared with its initial state or characteristics. The above parameters are therefore illustrative examples and not an exhaustive list.

[0125] In one embodiment, the concentrated milk product is resistant to: (a) sedimentation; (b) liquid phase separation; (c) discoloration; and / or (d) alterations in sensory profile, when stored.

[0126] Storage may refer to any suitable storage condition, including but not limited to storage - for any period of time - at ambient temperatures, such as about 20°C to about 25°C; cooled temperatures, such as about 4°C to 20°C; or above ambient temperatures.

[0127] In one embodiment, the concentrated milk product has an improved sensory profile as compared with a concentrated milk product with substantially the same proportion of total milk solids and / or milk proteins that has not been subjected to transglutaminase treatment. In one embodiment, the concentrated milk product has a sensory profile that is substantially the same as a concentrated milk product with a higher proportion of total milk solids and / or protein that has not been subjected to transglutaminase treatment.

[0128] In one embodiment, the concentrated milk product with a higher proportion of total milk solids and / or protein that has not been subjected to transglutaminase treatment has 0.5%, 1 %, 1.5%, 2% or more total milk solids and / or milk proteins than the concentrated milk product of the invention with comparable sensory characteristics.

[0129] The sensory profile referred to herein includes the perception of: (a) colour; (b) texture and / or consistency; (c) mouthfeel; (d) saltiness; (e) creaminess; and / or (f) overall taste.

[0130] The sensory characteristics may be based on sight, smell, and / or taste.

[0131] In one embodiment, the concentrated milk product is stable at:

[0132] (a) temperatures up to about 30°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 120 days, at least 140 days, at least 160 days, at least 180 days, at least 200 days, at least 220 days, at least 240 days, at least 260 days, at least 280 days, at least 300 days, at least 320 days, at least 340 days, or at least 360 days; and / or

[0133] (b) temperatures between about 21°C and about 25°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 120 days, at least 140 days, at least 160 days, at least 180 days, at least 200 days, at least 220 days, at least 240 days, at least 260 days, at least 280 days, at least 300 days, at least 320 days, at least 340 days, or at least 360 days.

[0134] (c) temperatures about 37°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, or at least 90 days, optionally wherein stability is determined by inspection.

[0135] Condensed milk, evaporated milk, and creamers

[0136] Concentrated milk products include condensed milks, evaporated milks, and creamers and are well known in the art. Moreover, standard methods of manufacture of such products are also well known. These methods may be adapted to comprise a method (i.e., the method steps) according to the present invention or to incorporate a bio-transformed precursor or biotransformed concentrated milk product according to the present invention The bio-transformation described herein facilitates the production of milk products with equivalent sensory profiles whilst using comparably lower amounts of total milk solids (and / or total milk protein).

[0137] Hence, the concentrated milk product of the invention may comprise a product as described in Table 1 , wherein the concentrated milk product has an improved sensory profile as compared with an equivalent non-bio-transformed product.

[0138] Preferably, the concentrated milk product of the invention has a lower amount of milk proteins and / or milk solids as compared with any value shown in Table 1 , when used in an equivalent product.

[0139] Suitably, the concentrated milk product of the invention has 0.1 %, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1 %, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.5%, 3%, or more than 3% less milk protein than a prior art concentrated milk product or milk product as described in Table 1.

[0140] Suitably, the concentrated milk product of the invention has 0.1 %, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1 %, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.5%, 3%, or more than 3% less total milk solids than a prior art concentrated milk product or milk product as described in Table 1.

[0141] Suitably, the concentrated milk product of the invention has 0.5 % to 1.5% less total milk solids, total milk fats, and / or total milk protein than a prior art concentrated milk product or milk product as described in Table 1.

[0142] Table 1 : Illustrative compositions of concentrated milk products

[0143] It will be understood that the values in table 1 are illustrative and other suitable values will be known. The definition of each product may vary due to the regulatory requirements in different jurisdictions; this will not impact the applicability of the present invention to such a product.

[0144] Suitably, the concentrated milk product of the invention may be an: evaporated milk, evaporated skimmed milk, evaporated partly skimmed milk, evaporated high-fat milk, evaporated milk with added vegetable lipids, condensed milk, condensed skimmed milk, condensed partly skimmed milk, condensed high-fat milk, a liquid creamer product, a powdered creamer product.

[0145] In one embodiment, the method of the present invention is a method for the manufacture of a condensed milk product, such as a condensed milk product described herein.

[0146] In one embodiment, the method of the present invention is a method for the manufacture of an evaporated milk product, such as an evaporated milk product described herein.

[0147] In one embodiment the concentrated milk product is a liquid product.

[0148] In one embodiment the concentrated milk product is a solid product, such as a powdered product.

[0149] Where the concentrated milk product is a powdered milk product the recited total milk solids and total milk protein values may refer to:

[0150] (a) total weight % of the dry weight of the formula; and / or

[0151] (b) total weight % of the weight of a liquid formula, such as a liquid formula prepared from the dry formula according to manufacturer instructions.

[0152] The bio-transformed precursors or concentrated milk products according to the invention may be used as the basis for the production of another concentrated milk product, such as wherein the advantageous properties of the cross-linked proteins demonstrated herein are also beneficial. In general, it is envisaged that the bio-transformed milk proteins will improve the sensory profiled and stability of the products to which they are added. The bio-transformed precursors or concentrated milk products according to the invention may be used as the basis for the production of a creamer product.

[0153] In one embodiment, the method of the present invention is a method for the manufacture of a creamer product.

[0154] Creamers may comprise from 0% milk fats to 30% milk fats, with optional vegetable lipids added up to a total lipid (i.e. , milk fats plus vegetable lipids) content of 30%.

[0155] The bio-transformed (i.e., cross-linked) milk proteins generated by the method of the present invention may be utilized in the manufacture of concentrated milk products, such as a creamer powder product or liquid creamer product.

[0156] Precursors

[0157] The present invention is concerned with concentrated milk products and their manufacture. As such, the present invention also concerns precursors of said products. In particular, the invention is concerned with precursors to which transglutaminase is added in order to effect an enzymatic change in the protein substituents of said precursor to thereby produce a biotransformed composition. The resultant bio-transformed composition may itself be a precursor (e.g., if it requires further processing that changes its composition) or may be considered to be the final concentrated milk product.

[0158] As used herein, the term precursor may refer to any composition (e.g., a liquid, or solid, such as a powder) that comprises milk proteins and / or other milk solids and is directly involved in the manufacture of a concentrated milk product according to the invention.

[0159] Precursors referred to herein may be bio-transformed (i.e., precursors present after a step of transglutaminase addition) or not bio-transformed (i.e., precursors present prior to a step of transglutaminase addition).

[0160] In one embodiment, the precursor comprises or consists of: (a) raw or untreated milk; (b) skimmed milk; (c) concentrated milk; (d) reconstituted milk powder; (e) evaporated milk; (f) reverse osmosis filtered milk; or (g) or a mixture of any of (a) to (f).

[0161] In one embodiment the milk is or is derived from Cow’s milk, Sheep’s milk, or Goat’s milk In one embodiment precursor has a milk protein content of at least 3% by weight, such as at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10% by weight.

[0162] In one embodiment the precursor has a milk protein content of at about 3% by weight to about 10% by weight, such as about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 9%, about 6% to about 8%, or about 6% to about 7% by weight.

[0163] In one embodiment the precursor has a total milk solids content of at least 15% by weight.

[0164] In one embodiment the precursor has a total milk solids content of at least 16% by weight.

[0165] In one embodiment the precursor has a total milk solids content of at least 17% by weight.

[0166] In one embodiment the precursor has a total milk solids content of at least 18% by weight.

[0167] In one embodiment the precursor has a total milk solids content of at least 19% by weight.

[0168] In one embodiment the precursor has a total milk solids content of at least 19.5% by weight.

[0169] In one embodiment the precursor has a total milk solids content of at least 20% by weight.

[0170] In one embodiment the precursor has a total milk solids content of at least 20.5% by weight.

[0171] In one embodiment the precursor has a total milk solids content of at least 21 % by weight.

[0172] In one embodiment the precursor has a total milk solids content of about 20% by weight.

[0173] In one embodiment the precursor has a total milk solids content of about 20.5% by weight.

[0174] In one embodiment the precursor has a total milk solids content of about 15% by weight, about 15.5% by weight, about 16% by weight, 16.5% by weight, about 17% by weight, about 17.5% by weight, about 18% by weight, about 18.5% by weight, about 19% by weight, about 19.5% by weight, about 20% by weight, about 20.5% by weight, about 21% by weight, about 21.5% by weight, or about 22% by weight. In one embodiment the precursor has a total milk solids content of at about 15% by weight to about 30% by weight, such as about 15% to about 25%, about 15% to about 24%, about 15% to about 23%, about 15% to about 22%, about 15% to about 21 %, about 15% to about 20%, about 17% to about 30%, about 17% to about 25%, about 17% to about 24%, about 17% to about 23%, about 17% to about 22%, about 17% to about 21%, about 17% to about 20%, about 19% to about 30%, about 19% to about 25%, about 19% to about 24%, about 19% to about 23%, about 19% to about 22%, about 19% to about 21%, about 19% to about 20%, about 20% to about 30%, about 20% to about 25%, about 20% to about 24%, about 20% to about 23%, about 20% to about 22%, or about 20% to about 21%.

[0175] In one embodiment the precursor has a total milk solids content of about 19% by weight to about 22% by weight.

[0176] In one embodiment the precursor has a total milk solids content of about 19% by weight to about 21 % by weight.

[0177] In one embodiment the precursor has a total milk solids content of about 20% by weight to about 21 % by weight.

[0178] In one embodiment the precursor has:

[0179] (a) a milk protein content of at about 3% by weight to about 10% by weight, such as about 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, about 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, about 5% to about 9%, about 5% to about 8%, about 5% to about 7%, about 5% to about 6%, about 6% to about 9%, about 6% to about 8%, or about 6% to about 7% by weight; and / or

[0180] (b) a total milk solids content of at about 15% by weight to about 30% by weight, such as about 15% to about 25%, about 15% to about 24%, about 15% to about 23%, about 15% to about 22%, about 15% to about 21%, about 15% to about 20%, about 17% to about 30%, about 17% to about 25%, about 17% to about 24%, about 17% to about 23%, about 17% to about 22%, about 17% to about 21%, about 17% to about 20%, about 19% to about 30%, about 19% to about 25%, about 19% to about 24%, about 19% to about 23%, about 19% to about 22%, about 19% to about 21%, about 19% to about 20%, about 20% to about 30%, about 20% to about 25%, about 20% to about 24%, about 20% to about 23%, about 20% to about 22%, or about 20% to about 21%. In one embodiment, the precursor has a total lipid content of about 1 % to about 10% by weight, such as about 1 % to about 9%, about 1 % to about 8%, about 1 % to about 7%, about 1 % to about 6%, about 1 % to about 5%, about 1% to about 4%, about 1 % to about 3%, about 1% to about 2%, 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, 5% to about 9%, about 5% to about 8%, about 5% to about 7%, or about 5% to about 6%.

[0181] In one embodiment, the precursor has a total milk lipid and / or fat content of about 1 % to about 10% by weight, such as about 1 % to about 9%, about 1 % to about 8%, about 1 % to about 7%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%, about 1 % to about 3%, about 1% to about 2%, 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, about 2% to about 3%, 3% to about 9%, about 3% to about 8%, about 3% to about 7%, about 3% to about 6%, about 3% to about 5%, about 3% to about 4%, 4% to about 9%, about 4% to about 8%, about 4% to about 7%, about 4% to about 6%, about 4% to about 5%, 5% to about 9%, about 5% to about 8%, about 5% to about 7%, or about 5% to about 6%.

[0182] In one embodiment, the precursor has a total lipid content of about 4% to about 5% by weight.

[0183] The precursor may be a bio-transformed precursor or a non-bio-transformed precursor.

[0184] In one embodiment, the precursor according to the preceding embodiments is a biotransformed precursor (i.e. , subsequent to transglutaminase addition).

[0185] In one embodiment, the precursor according to the preceding embodiments is a non-bio- transformed precursor (i.e., prior to transglutaminase addition).

[0186] In one embodiment, the precursor is the precursor to which transglutaminase is added (i.e., directly).

[0187] In one embodiment, provided herein in a bio-transformed precursor of a milk product, produced according to the present invention. The bio-transformed precursor of a milk product, produced according to the present invention may for example undergo further processing to extract certain components, such as lactose e.g., in order to produce creamer products. Further, the bio-transformed precursor may be processed into a powder that can be reconstituted into a concentrated milk product according to the invention.

[0188] Transglutaminase

[0189] Transglutaminases are a class of enzymes that catalyze the formation of a covalent bond between the side-chain amide (-CO-NH2) of a glutamine (Q) residue and the side-chain amine of a lysine (K).

[0190] In the context of polypeptides and proteins, the activity of transglutaminase enzymes can promote the formation of covalent cross-links between distinct polypeptide chains (e.g., of the same or different identity; intermolecular cross-linking) or within a single polypeptide (i.e., intramolecular or intermolecular cross-linking).

[0191] In the context of the present invention a transglutaminase is considered to be any protein or polypeptide with transglutaminase activity, i.e., capable of cross-linking proteins / polypeptides as described above.

[0192] The activity of transglutaminase on target proteins may be assessed by various methods known in the art. For example, SDS-PAGE analysis wherein mass shifting of protein substituents - as compared to an untreated sample - to higher molecular weight regions of the gel are indicative of an increase in unimolecular molecular mass as a result of the formation of new covalent bonds between proteins (i.e., cross-linking). Other methods such as mass- spectrometry may also be used to assess the nature and extent of cross-linking.

[0193] For example, cross-linking may be assessed on the basis of a change in biophysical parameters (e.g., size) of any of: whey proteins (e.g., p-lactoglobulin, a-lactalbumin, glycomacropeptide, serum albumin, immunoglobulins), caseins (e.g., a-s1-casein, a-s2- casein, p-casein, K-casein, and / or y-casein), or any other milk protein.

[0194] The native parameters of milk proteins are well studied and thus any size change induced by cross-linking may be easily identified by the person skilled in the art. Other molecular mass / size based analysis techniques may be used as appropriate, such as analytical (size-exclusion) chromatography, analytical ultra-centrifugation (AUG), and / or dynamic light scattering (DLS).

[0195] The transglutaminase of the present invention may be obtained from any suitable source (e.g., organism).

[0196] In one embodiment, the transglutaminase is a microbial transglutaminase.

[0197] In one embodiment, the transglutaminase is a bacterial transglutaminase.

[0198] In one embodiment, the transglutaminase a Streptomyces mobaraensis transglutaminase.

[0199] In one embodiment, the transglutaminase is a food-grade transglutaminase.

[0200] Transglutaminase addition

[0201] Addition of the transglutaminase may be performed at any suitable step of any of the methods according to the present invention.

[0202] Suitably, the transglutaminase is added to the entire batch of precursor (e.g., as opposed to a sample taken from the precursor of the invention).

[0203] In some embodiments the addition of transglutaminase may be timed such that it is performed during a step in a pre-existing process (e.g., the wet-mixing of ingredients) such that the overall manufacturing process is not subject to an increase in process time. This may have the advantage of, for example, not increasing costs and / or reducing efficiency.

[0204] In one embodiment, the transglutaminase is added at about 0.5% to about 5% by weight of the total protein content of the precursor, such as about 0.5% to about 2%, about 1% to about 2%, or about 1.5% to about 2% by weight of the total protein content of the precursor.

[0205] In one embodiment, the transglutaminase is added at about 0.5% to about 5% by weight of the total protein content of the precursor, such as about 0.5% to about 2%, about 1% to about 2%, or about 1.5% to about 2% by weight of the total protein content of the precursor, wherein the enzyme activity is about 100 to about 140 ll / g. In one embodiment, the transglutaminase is added in an amount of at least about 0.5% by weight of the total protein content of the precursor, such at least about 0.6%, 0.7%, 0.8%, 0.9%, 1 %, 1 .5%, 2%, 2.5%, or at least about 3% by weight of the total protein content of the precursor.

[0206] For example, where the transglutaminase is added at 1 .5% by weight of total protein, if a 100g precursor comprises 5g of protein, 0075g of transglutaminase is to be added.

[0207] In another embodiment, the transglutaminase is added at about 1.5% by weight of the total protein content of the precursor.

[0208] In one embodiment, the transglutaminase added to a precursor that has a milk protein content of at least 4%, such as at least 5%.

[0209] In one embodiment, the transglutaminase added to a precursor that has a milk protein content of about 3% to about 6%, such as 3.5%, 4%, 4.5%, 5%, or 5.5%.

[0210] In one embodiment, the transglutaminase added to a precursor that has a milk protein content of about 5%.

[0211] In one embodiment the transglutaminase is added in an amount of about 50 II (units of enzyme activity) per 100 grams of protein in the precursor to about 300 II per 100g of protein in the precursor, such about 60 U to 290 U, 70 U to 280 U, 80 U to 270 U, 90 U to 260U, 100 U to 250U, 110 U to 240 U, 120 U to 230 U, 130 U to 220 U, 140 U to 210 U, 140 U to 210 U, or 150 II to 200 II per gram of protein in the precursor.

[0212] In one embodiment the transglutaminase is added in an amount of about 140 II per 100g of protein in the precursor to about 210 II per 100g of protein in the precursor.

[0213] In one embodiment the transglutaminase is added in an amount of at least 50 II (units of enzyme activity) per 100 grams of protein in the precursor, such as at least 60 II, 70 II, 80 II, 90 U, 100 U, 110 U, 120 U, 130 U, 140 U, 150 U, 160 U, 170 U, 180 U, 190 U, 200 U, 210 U, 220 U, 230 U, 240 U, 250 U, 260 U, 270 U, 280 U, 290 U, or at least 300 U per gram of protein in the precursor.

[0214] Determination of enzyme activity is well known to the person skilled in the art and can be performed using any appropriate assay. The amount of transglutaminase added may be altered. For example, the amount of enzyme may be reduced if the activity of the enzyme is particularly high, or, for example if the amount of time the enzyme is in contact with the precursor is extended (e.g., a longer reaction time).

[0215] In the context of the present invention, any milk product or precursor thereof may be considered a bio-transformed milk product when acted upon by transglutaminase, that is, comprising any amount of cross-linking between substituent proteins.

[0216] Following transglutaminase addition to a precursor it may not be essential to allow a designated ‘incubation’ step since the enzyme will work to varying degrees in a variety of favourable conditions. Such conditions may already be present in a ‘normal’ work-flow for producing concentrated milk products.

[0217] The transglutaminase may be added concomitantly with one or more preexisting steps in a method for production of a concentrated milk product. For example, wherein the one or more preexisting steps is:

[0218] (a) Wet mixing of ingredients, such as when the precursor comprises recombined powdered ingredients;

[0219] (b) Any step following evaporation but prior to sterilization; and / or

[0220] (c) Following standardization of the product.

[0221] The transglutaminase may be added before, after, or concomitantly with any one or more of the steps shown in Figure 1.

[0222] In one embodiment, the transglutaminase is added to a precursor that has been subjected to heat treatment (e.g., pasteurization).

[0223] In one embodiment, the transglutaminase is added to a precursor that has been subjected to homogenization.

[0224] In one embodiment, the transglutaminase is added to a precursor that has been subjected to heat treatment (e.g., pasteurization) and homogenization.

[0225] In some embodiments the precursor is maintained under specific conditions following transglutaminase activity. In one embodiment, the precursor is maintained at a temperature of about 40°C to about 60°C, such as about 45°C to about 60°C, about 50°C to about 60°C, or about 55°C to about 60°C, following addition of the transglutaminase.

[0226] In one embodiment, the precursor is maintained at a temperature of about 55°C to about 60°C, following addition of the transglutaminase.

[0227] In one embodiment, the precursor is maintained at a temperature of about 50°C to about 60°C, following addition of the transglutaminase at about 1 % to 2% by weight of the total weight of protein, optionally wherein protein content of the precursor is about 3% to about 6%.

[0228] In one embodiment, the precursor is maintained at a temperature of about 55°C to about 60°C, following addition of the transglutaminase at about 1.5% by weight of the total weight of protein, optionally wherein protein content of the precursor is about 5%.

[0229] In one embodiment, the precursor is maintained at a temperature according to any of the foregoing embodiments for at least 10 minutes, such as at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 65 minutes, at least 70 minutes, at least 75 minutes, at least 80 minutes, or longer, subsequent to the addition of the transglutaminase.

[0230] Subsequent to its addition, the activity of the transglutaminase may be controlled e.g., by altering the temperature of the precursor. It will be understood that both high and low temperatures will reduce, inhibit, or terminated the activity of the transglutaminase due to e.g., altered enzyme kinetics and eventual irreversible denaturation (and loss of function).

[0231] In one embodiment activity of the transglutaminase is terminated, reduced, and / or inhibited subsequent to its addition.

[0232] In one embodiment activity of the transglutaminase is terminated, reduced, and / or inhibited by cooling the bio-transformed precursor and transglutaminase mixture to a temperature of less than about 40°C, such as less than about 30°C, less than about 20°C, less than about 10°C, or less than about 5°C. In one embodiment activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of at least about 70°C, such as at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, at least about 120°C, at least about 130°C, or at least about 140°C.

[0233] By way of example, the present inventors found that transglutaminase inactivation is achieved by heating the enzyme-precursor mixture under the following conditions:

[0234] • 75°C for 20 seconds;

[0235] • 75°C for 60 seconds;

[0236] • 85°C for 60 seconds;

[0237] • 90°C for 20 seconds;

[0238] • 90°C for 60 seconds;

[0239] In one embodiment, activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of at least 75°C for at least 20 seconds.

[0240] In one embodiment, activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of about 75°C to about 90°C for about 20 to about 60 seconds.

[0241] In one embodiment, activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of about 75°C for about 20 to about 60 seconds.

[0242] In one embodiment, activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of about 85°C for about 20 to about 60 seconds.

[0243] In one embodiment, activity of the transglutaminase is terminated, reduced, and / or inhibited by heating the bio-transformed precursor and transglutaminase mixture to a temperature of about 90°C for about 20 to about 60 seconds. Methods for production of a bio-transformed concentrated milk product

[0244] Provided herein is a method for the production / manufacture of a concentrated milk product, in particular a bio-transformed concentrated milk product, and other bio-transformed milk product precursors.

[0245] In one aspect, there is provided a method for the production of a concentrated milk product having:

[0246] (a) a milk protein content of at least 3.5% by weight; and / or

[0247] (b) a total milk solids content of at least 15% by weight; wherein the method comprises the step of adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product.

[0248] In one embodiment, the precursor has:

[0249] (a) a milk protein content of at least about 4% by weight, such as at least about 5% by weight; and / or

[0250] (b) a total milk solids content of about 13% by weight to about 40% by weight, preferably about 20% by weight to about 25% by weight, more preferably about 23% by weight to about 24% by weight.

[0251] In one embodiment, the transglutaminase is added:

[0252] (a) at about 0.5% to about 5% by weight of the total protein content of the precursor, such as about 0.5% to about 2%, about 1 % to about 2%, or about 1.5% to about 2% by weight of the total protein content of the precursor; and / or

[0253] (b) in an amount of about 50 II to about 300 II per 100 grams of protein in the precursor, such as about 140 II to 210 II per 100g of protein.

[0254] In one embodiment, the transglutaminase is added to a precursor:

[0255] (a) at about 0.5% to about 5% by weight of the total protein content of the precursor, such as about 0.5% to about 2%, about 1 % to about 2%, or about 1.5% to about 2% by weight of the total protein content of the precursor; and / or

[0256] (b) in an amount of about 50 II to about 300 II per 100 grams of protein in the precursor, such as about 140 II to 210 II per 100g of protein,

[0257] Wherein the precursor has a milk protein content of about 3% to about 6%, such as 3.5%, 4%, 4.5%, 5%, or 5.5%.

[0258] In one embodiment, the precursor is maintained at a temperature of about 40°C to about 60°C, such as about 45°C to about 60°C, about 50°C to about 60°C, preferably about 55°C to about 60°C, subsequent to the addition of the transglutaminase; optionally wherein the precursor is maintained at said temperature for at least 10 minutes, such as at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 65 minutes, at least 70 minutes, at least 75 minutes, at least 80 minutes, or longer, subsequent to the addition of the transglutaminase.

[0259] In one embodiment, the transglutaminase is added to a precursor wherein the milk protein content is about 5%, the temperature of the precursor is about 55°C, and wherein the transglutaminase is added at about 1 .5% by weight of the total protein content of the precursor.

[0260] The methods of the invention may comprise a step of terminating or reducing (i.e., controlling) the transglutaminase reaction.

[0261] In one embodiment, the method additionally comprises the step of terminating, reducing, and / or inhibiting the activity of the transglutaminase, subsequent to the addition of the transglutaminase.

[0262] In one embodiment, the terminating, reducing, and / or inhibiting step comprises cooling the bio-transformed precursor and transglutaminase mixture to a temperature of less than about 40°C, such as less than about 30°C, less than about 20°C, less than about 10°C, or less than about 5°C.

[0263] In another embodiment, the terminating, reducing, and / or inhibiting step comprises heating the bio-transformed precursor and transglutaminase mixture to a temperature of at least about 70°C, such as at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, at least about 120°C, at least about 130°C, or at least about 140°C.

[0264] The terminating, reducing, and / or inhibiting step may be a pre-existing step in a process or a step that is also required for an alternative (and additional) purpose. For example, a standard sterilization process would also serve to inactive the transglutaminase.

[0265] In one embodiment, the method comprises the step of heat treatment and / or sterilization of the bio-transformed precursor by heating to at least about 120°C, such as at least about 125°C, at least about 130°C, at least about 135°C, or at least about 140 °C. In one embodiment, the heat treatment and / or sterilization comprises or consists of:

[0266] (a) retort sterilization;

[0267] (b) direct steam injection sterilization; and / or

[0268] (c) indirect sterilization.

[0269] In one embodiment, the step of transglutaminase addition is performed during wet mixing of ingredients (e.g., the reconstitution of milk powder).

[0270] In one embodiment, the step of transglutaminase addition is performed subsequent to heat treatment or pasteurization of the precursor.

[0271] In one embodiment, the step of transglutaminase addition is performed subsequent to homogenization of the precursor.

[0272] In one embodiment, the step of transglutaminase addition is performed subsequent to standardization of the precursor.

[0273] In one embodiment, the step of transglutaminase addition is performed prior to sterilization.

[0274] In one embodiment, the step of transglutaminase addition is performed prior to canning / packaging.

[0275] Certain processes may alter the structures (e.g., micellar structures or other quaternary structures) of milk proteins and thus alter the availability and / or accessibility of certain milk proteins as substrates for transglutaminase enzymes.

[0276] The present inventors have found that it is particularly advantageous to perform the biotransformation, i.e., add transglutaminase, following heat treatment (e.g., pasteurization) and / or homogenization of the precursor.

[0277] Bio-transformation of the concentrated milk product precursors following heat treatment and / or homogenization yields a particularly stable and consistent product.

[0278] In one embodiment heat treatment and / or pasteurization comprises heating the precursor to at least about 60°C, such as at least about 70°C, at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, or at least about 120°C. In one embodiment heat treatment and / or pasteurization comprises heating the precursor to any of the above temperatures and maintaining it at said temperature for at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, or at least 5 minutes.

[0279] In one embodiment, the step of transglutaminase addition is performed subsequent to heat treatment / pasteurization of the precursor and homogenization, wherein said heat treatment and / or pasteurization comprises heating the precursor to at least about 60°C, such as at least about 70°C, at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, or at least about 120°C, optionally wherein said temperature is maintained for at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, or at least 5 minutes.

[0280] In one embodiment, the step of transglutaminase addition is performed subsequent to heat treatment / pasteurization of the precursor and homogenization, wherein said heat treatment and / or pasteurization comprises heating the precursor to about 120°C and maintaining said temperature for about 2 minutes.

[0281] In one embodiment the method is an industrial process.

[0282] In one embodiment, the total volume of concentrated milk product, precursor and / or biotransformed precursor is at least 100 liters, such as at least 200 liters, at least 300 liters, at least 400 liters, at least 500 liters, at least 1000 liters, at least 5000 liters, at least 10000 liters, at least 15000 liters, at least 20000 liters, at least 25000 liters, at least 30000 liters, at least 35000 liters, at least 40000 liters, at least 45000 liters, at least 50000 liters, at least 100000 liters, at least 150000 liters, or at least 200000 liters.

[0283] In one embodiment, the total volume of concentrated precursor to which the transglutaminase is added is at least 100 liters, such as at least 200 liters, at least 300 liters, at least 400 liters, at least 500 liters, at least 1000 liters, at least 5000 liters, at least 10000 liters, at least 15000 liters, at least 20000 liters, at least 25000 liters, at least 30000 liters, at least 35000 liters, at least 40000 liters, at least 45000 liters, at least 50000 liters, at least 100000 liters, at least 150000 liters, or at least 200000 liters.

[0284] In one embodiment, the method comprises the step of adding carrageenan to the precursor and / or the bio-transformed precursor.

[0285] In one embodiment, the method comprises the step of adding phosphate to the biotransformed precursor. In one embodiment, the amount of phosphate added to the bio-transformed precursor is 0.01 % to 0.3% by weight.

[0286] In one embodiment, the amount of phosphate added to the bio-transformed precursor is less than 0.16%.

[0287] In one embodiment, there is provided a method for the production of a concentrated milk product wherein the method comprises the step of:

[0288] (a) adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product, wherein:

[0289] (i) the precursor has a milk protein content of about 4% by weight to about 6% by weight, preferably 5% by weight;

[0290] (ii) the transglutaminase is added at 1.5% by weight as a percentage of the weight of the total milk protein content of the precursor; and / or

[0291] (iii) optionally, wherein the precursor is at about 50°C to about 60°C, preferably about 55°C;

[0292] (b) heating the transglutaminase enzyme and precursor mixture to- or maintaining the transglutaminase enzyme and precursor mixture at about 50°C to about 60°C, preferably about 55°C;

[0293] (c) maintaining the temperature of step (b) for at least 40 minutes, preferably at least 50 minutes;

[0294] (d) optionally, cooling the bio-transformed precursor to below about 10°C, such as below about 8°C;

[0295] (f) optionally, terminating the activity of the transglutaminase by heating the biotransformed precursor and transglutaminase mixture to a temperature of at least 75°C for at least 20 seconds; and

[0296] (g) heat treatment and / or sterilization of the bio-transformed precursor comprising heating the precursor to about 120°C and maintaining said temperature for at least 30 seconds.

[0297] In one embodiment, there is provided a method for the production of a concentrated milk product having:

[0298] (a) a milk protein content of at least 3.5% by weight; and / or

[0299] (b) a total milk solids content of at least 15% by weight; wherein the method comprises the step of: (c) adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product, wherein the precursor has a milk protein content of about 4% by weight to about 6% by weight, preferably 5% by weight; and wherein the transglutaminase is added at 1 .5% by weight as a percentage of the weight of the total milk protein content of the precursor, optionally wherein the precursor is at about 50°C to about 60°C, preferably about 55°C;

[0300] (d) heating the transglutaminase enzyme and precursor mixture to- or maintaining the transglutaminase enzyme and precursor mixture at about 50°C to about 60°C, preferably about 55°C at maintaining said temperature for at least 40 minutes, preferably at least 50 minutes;

[0301] (e) optionally, cooling the bio-transformed precursor to below about 10°C, such as below about 8°C;

[0302] (f) optionally, terminating the activity of the transglutaminase by heating the biotransformed precursor and transglutaminase mixture to a temperature of about 75°C to about 90°C for about 20 to about 60 seconds; and

[0303] (g) heat treatment and / or sterilization of the bio-transformed precursor comprising heating the precursor to about 120°C and maintaining said temperature for at least 30 seconds.

[0304] Other steps may be combined with those described above.

[0305] Unless otherwise stated, or made obvious by the context of the disclosure, all % values herein (e.g., with reference to milk solids and / or protein) are % by weight values.

[0306] All publications mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described methods and system of the 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 molecular biology or related fields are intended to be within the scope of the following claims. EXAMPLES

[0307] The invention will now be described by way of the following non-limiting examples.

[0308] Example 1 - Illustrative work-flow for the manufacture of concentrated milk products

[0309] The present invention is compatible with numerous manufacturing processes or work-flows. Figure 1 illustrated a generalized example work-flow that is compatible with the methods of the present invention.

[0310] Example 2 - Bio-transformation facilitates a reduction in milk solids

[0311] The ability to integrate bio-transformation of concentrated milk products and their precursors into work-flows was assessed and the characteristics of the resultant products analyzed.

[0312] Figure 2 illustrates an example work flow for the preparation of bio-transformed concentrated milk products such as those tested in Example 2. By way of summary, samples were prepared with the following parameters:

[0313] • Standardization using fresh milk, skim milk powder, skim milk liquid & milk fat anhydrous. Starting around 13% total milk solids (TS)

[0314] • 1stheat treatment: 110°C / 120sec

[0315] • Evaporation to 23% Total Solids

[0316] • Homogenization: 180bar + 40 bar at 75°C

[0317] • Enzymatic reaction by batch: o 55°C for 50 min (cooled to 8°C) o Enzyme dose was 2% / protein.

[0318] • Final standardization to TS target (below) with water addition

[0319] • Sterilization in cans

[0320] • Indirect UHT treatment follow by aseptic homogenization.

[0321] • Direct UHT treatment follow by aseptic homogenization.

[0322] The precursor to which transglutaminase was added comprised total milk solids of about 22% and total protein of about 5%.

[0323] The resultant concentrated milk products were standardized to produce two different groups of products:

[0324] (1) a reference (not bio-transformed) having about 21 .8% total milk solids, about 5.5% fat, and about 5.7% milk protein. (2) a bio-transformed sample having about 20.5% total milk solids, about 5.2% fat, and about 5.2% milk protein.

[0325] The Sensory profile of each group was analysed. The sensory profile was similar between reference and bio-transformed samples. Surprisingly, no perceived decrease in mouthfeel was observed in the bio-transformed samples despite a reduction in total solids content.

[0326] Reference and bio-transformed samples were also tested in cooking (creamed spinach, pasta, and rice pudding) and for other applications such as a coffee creamer etc. Again, it was observed that bio-transformed samples had no reduction in sensory performance as compared with reference despite a reduced total solids content.

[0327] Hence, it is evident that bio-transformation according to the invention allows the production of bio-transformed concentrated milk products that have similar or the same sensory characteristics as equivalent non-bio-transformed concentrated milk products that comprise higher quantities of milk solids. In other words, the methods and products of the invention facilitate a reduction in total milk solids whilst retaining culinary performance.

[0328] Bio-transformation also improves stability. Retort sterilized samples, i.e., samples sterilized in cans (see Figure 2), demonstrated improved stability of the bio-transformed product over the non-bio-transformed reference product insofar as the former was not as prone to sedimentation as the latter (see Table 2).

[0329] Table 2: milk product storage characteristics

[0330] Sedimentation was assessed and assigned a numerical category identified based on severity.

[0331] As illustrated in Table 2, Biotransformation improves the shelf life by decreasing sediment.

[0332] Example 3 - Bio-transformation improves stability

[0333] In view of the findings of improved stability evidenced by Example 2, further stability tests were performed.

[0334] The temporal stability of bio-transformed products was assessed using the same ‘quality keeping test’ (QKT). Visual inspection bio-transformed and non-bio-transformed concentrated milk products was carried out on samples that had been stored under varying conditions in order to assess phase separation, in particular the precipitation of solid sediment in the container (Table 3).

[0335] Sedimentation is an undesirable characteristic for consumers.

[0336] Two independent concentrated milk products were tested:

[0337] • Sample 1 was an evaporated milk product comprising 21.8% total milk solids (i.e., non- bio-transformed).

[0338] • Sample 2 was a transglutaminase treated evaporated milk product comprising 20.5% total milk solids (i.e., bio-transformed).

[0339] Both samples were subject to Indirect ultra heat treatment (UHT) at 138.5°C for 30s in order to sterilize the product before packaging. The packaged products were then stored under identical conditions.

[0340] Table 3: milk product storage characteristics

[0341] The corresponding visual results are reproduced in Figure 3.

[0342] Sedimentation was assessed and assigned a numerical category identified based on severity.

[0343] The bio-transformed product exhibited no sedimentation during storage, indicative of improved stability. The bio-transformed product therefore exhibited improved stability and desirable storage characteristics (i.e., no sedimentation over time).

[0344] Example 4 - Increasing pH after bio-transformation improves product stability

[0345] Heat stability of concentrated milk products may be altered by the addition of stabilizing agents such as phosphates at one or more standardization steps.

[0346] However, the addition of phosphates can be associated with an increased - and undesirable - perception of saltiness.

[0347] The present inventors thus sought to determine whether the requirement for phosphates could be reduced.

[0348] Figure 4 illustrates an example work flow for the preparation of bio-transformed concentrated milk products such as those tested herein. As can be seen, a set of 5 different bio-transformed milk products were standardized at different pH levels in order to assess their phosphate requirement and stability.

[0349] The inventors found that increasing the pH of the bio-transformed precursor required the addition of lower amounts of phosphates in order to provide heat stability (e.g., stability of the product when subjected to a sterilization procedure).

[0350] Hence, increasing the pH of the bio-transformed milk product improves heat stability (including resistance to sterilization temperatures) and reduces the need for phosphate addition, thereby also reducing saltiness. EMBODIMENTS

[0351] The invention will now be described by way of the following numbered embodiments.

[0352] Embodiment 1 . A method for the production of a concentrated milk product having:

[0353] (a) a milk protein content of at least 3.5% by weight; and / or

[0354] (b) a total milk solids content of at least 15% by weight; wherein the method comprises the step of adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product.

[0355] Embodiment 2. The method of embodiment 1 , wherein the transglutaminase is a microbial transglutaminase, preferably a bacterial transglutaminase.

[0356] Embodiment s. The method of embodiment 1 or embodiment 2, wherein the transglutaminase is added:

[0357] (a) at about 0.5% to about 5% by weight of the total protein content of the precursor, such as about 0.5% to about 2%, about 1% to about 2%, or about 1 .5% to about 2% by weight of the total protein content of the precursor; and or

[0358] (b) in an amount of about 50 II to about 300 II per 100 grams of protein in the precursor, such as about 140 II to 210 II per 100g of protein.

[0359] Embodiment 4. The method of any of embodiments 1 to 3, wherein the precursor is maintained at a temperature of about 40°C to about 60°C, such as about 45°C to about 60°C, about 50°C to about 60°C, preferably about 55°C to about 60°C, following addition of the transglutaminase.

[0360] Embodiment 5. The method of embodiment 4, wherein the precursor is maintained at said temperature for at least 10 minutes, such as at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 35 minutes, at least 40 minutes, at least 45 minutes, at least 50 minutes, at least 55 minutes, at least 60 minutes, at least 65 minutes, at least 70 minutes, at least 75 minutes, at least 80 minutes, or longer, subsequent to the addition of the transglutaminase.

[0361] Embodiment 6. The method of any of embodiments 1 to 5, wherein the step of transglutaminase addition is performed when the milk protein content of the precursor is at least 4%, such as at least 5%. Embodiment ?. The method of any of embodiments 1 to 6, wherein the step of transglutaminase addition is performed concomitantly with one or more preexisting steps in the method for production of the concentrated milk product, optionally wherein said one or more preexisting steps is:

[0362] (a) wet mixing of ingredients, such as when the precursor comprises recombined powdered ingredients;

[0363] (b) any step following evaporation but prior to sterilization; and / or

[0364] (c) following standardization of the product.

[0365] Embodiment 8. The method of any of embodiments 1 to 7, wherein the method additionally comprises the step of terminating, reducing, and / or inhibiting the activity of the transglutaminase, subsequent to the addition of the transglutaminase.

[0366] Embodiment 9. The method of embodiment 8, wherein said terminating, reducing, and / or inhibiting step comprises:

[0367] (a) cooling the bio-transformed precursor and transglutaminase mixture to a temperature of less than about 40°C, such as less than about 30°C, less than about 20°C, less than about 10°C, or less than about 5°C; and / or

[0368] (b) heating the bio-transformed precursor and transglutaminase mixture to a temperature of at least about 70°C, such as at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, at least about 120°C, at least about 130°C, or at least about 140°C.

[0369] Embodiment 10. The method of any of embodiments 1 to 9, wherein the precursor comprises or consists of cow’s milk.

[0370] Embodiment 11. The method of any of embodiments 1 to 10, wherein the precursor comprises or consists of:

[0371] (a) raw or untreated milk;

[0372] (b) skimmed milk;

[0373] (c) concentrated milk;

[0374] (d) reconstituted milk powder;

[0375] (e) evaporated milk;

[0376] (f) reverse osmosis filtered milk; or

[0377] (g) or a mixture of any of (a) to (f). Embodiment 12. The method of any of embodiments 1 to 11 , wherein the precursor has a total milk solids content of about 13% by weight to about 40% by weight, preferably about 20% by weight to about 25% by weight, more preferably about 23% by weight to about 24% by weight.

[0378] Embodiment 13. The method of any of embodiments 1 to 12, wherein the precursor comprises carrageenan.

[0379] Embodiment 14. The method of any of embodiments 1 to 13, wherein the precursor is heat treated and / or pasteurized before transglutaminase addition.

[0380] Embodiment 15. The method of embodiment 14, wherein said heat treatment and / or pasteurization comprises heating the precursor to at least about 60°C, such as at least about 70°C, at least about 80°C, at least about 90°C, at least about 100°C, at least about 110°C, or at least about 120°C, optionally wherein said temperature is maintained for at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, or at least 5 minutes.

[0381] Embodiment 16. The method of any of embodiments 1 to 16, wherein the total volume of concentrated milk product, precursor and / or bio-transformed precursor is at least 100 liters, such as at least 200 liters, at least 300 liters, at least 400 liters, at least 500 liters, at least 1000 liters, at least 5000 liters, at least 10000 liters, at least 15000 liters, at least 20000 liters, at least 25000 liters, at least 30000 liters, at least 35000 liters, at least 40000 liters, at least 45000 liters, at least 50000 liters, at least 100000 liters, at least 150000 liters, or at least 200000 liters.

[0382] Embodiment 17. The method of any of embodiments 1 to 16, wherein the pH of the biotransformed precursor and / or milk product is pH 6.1 or above.

[0383] Embodiment 18. The method of any of embodiments 1 to 17, wherein the pH of the biotransformed precursor and / or milk product is pH 6.4 or above.

[0384] Embodiment 19. The method of any of embodiments 1 to 18, wherein the method comprises the step of adding stabilizing agent to the bio-transformed precursor.

[0385] Embodiment 20. The method of embodiment 19, wherein the stabilizing agent is a phosphate, optionally wherein the phosphate is:

[0386] (a) a sodium phosphate; (b) a potassium phosphate; or

[0387] (a) a mixture of sodium phosphate and potassium phosphate.

[0388] Embodiment 21. The method of any of embodiments 1 to 20, wherein the method comprises the step of heat treatment and / or sterilization of the bio-transformed precursor by heating to at least about 120°C, such as at least about 125°C, at least about 130°C, at least about 135°C, or at least about 140 °C.

[0389] Embodiment 22. The method of embodiment 21 , wherein:

[0390] (a) the method does not comprise an inactivation step according to embodiment 8 or embodiment 9; or

[0391] (b) the heat treatment and / or sterilization is performed in addition to the inactivation step as defined by embodiment 8 or embodiment 9.

[0392] Embodiment 23. The method of any of embodiments 1 to 22, wherein any of the heat treatment and / or sterilization steps comprises or consists of:

[0393] (a) retort sterilization;

[0394] (b) direct steam injection sterilization; and / or

[0395] (c) indirect sterilization.

[0396] Embodiment 24. The method of any of embodiments 1 to 23, wherein the concentrated milk product has a total milk solids content about 20% by weight to about 21% by weight, more preferably about 20.5%.

[0397] Embodiment 25. A concentrated milk product produced by the method of any of embodiments 1 to 24.

[0398] Embodiment 26. A concentrated milk product, wherein the concentrated milk product has:

[0399] (a) a milk protein content of at least 3.5% by weight; and / or

[0400] (b) a total milk solids content of about 15% by weight to about 26% by weight; and,

[0401] (c) cross-linked milk proteins; and optionally,

[0402] (d) trace amounts of an inactivated or non-functional transglutaminase; and / or

[0403] (e) carrageenan;

[0404] Preferably, where both (d) and (e) are present. Embodiment 27. The concentrated milk product of embodiment 25 or embodiment 26, wherein the milk product comprises 0.01 % to 0.3% by weight phosphates.

[0405] Embodiment 28. The concentrated milk product of any of embodiments 25 to 27, wherein the milk product comprises 0.02% to 0.2% phosphates.

[0406] Embodiment 29. The concentrated milk product of any of embodiments 25 to 28, wherein the concentrated milk product has a total milk solids content of about 20.5% by weight.

[0407] Embodiment 30. The concentrated milk product of any of embodiments 25 to 29, wherein the concentrated milk product has a total lipid content of about 4% to about 5% by weight.

[0408] Embodiment 31 . The concentrated milk product of any of embodiments 25 to 30, wherein the concentrated milk product has a total milk protein content of about 3% to about 6% by weight.

[0409] Embodiment 32. The concentrated milk product of any of embodiments 25 to 31 , wherein the concentrated milk product is lactose free or substantially lactose free.

[0410] Embodiment 33. The concentrated milk product of any of embodiments 25 to 32, wherein the concentrated milk product is not a fermented milk product.

[0411] Embodiment 34. The concentrated milk product of any of embodiments 25 to 33, wherein the concentrated milk product is:

[0412] (a) an evaporated milk product;

[0413] (a) a condensed milk product; and / or

[0414] (a) a creamer product;

[0415] Embodiment 35. The concentrated milk product of any of embodiments 25 to 33, wherein the concentrated milk product has:

[0416] (a) an improved sensory profile as compared with a concentrated milk product with substantially the same proportion of total milk solids that has not been subjected to transglutaminase treatment; and / or

[0417] (b) a sensory profile that is substantially the same as a concentrated milk product with a higher proportion of total milk solids that has not been subjected to transglutaminase treatment. Embodiment 36. The concentrated milk product of embodiment 35, wherein said sensory profile includes:

[0418] (a) perception of mouthfeel;

[0419] (b) perception of saltiness;

[0420] (c) perception of creaminess; and / or

[0421] (d) perception of overall taste.

[0422] Embodiment 37. The concentrated milk product of any of embodiments 25 to 36, wherein the concentrated milk product is stable at:

[0423] (a) temperatures up to about 30°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 120 days, at least 140 days, at least 160 days, at least 180 days, at least 200 days, at least 220 days, at least 240 days, at least 260 days, at least 280 days, at least 300 days, at least 320 days, at least 340 days, or at least 360 days; and / or

[0424] (b) temperatures between about 21°C and about 25°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, at least 90 days, at least 100 days, at least 120 days, at least 140 days, at least 160 days, at least 180 days, at least 200 days, at least 220 days, at least 240 days, at least 260 days, at least 280 days, at least 300 days, at least 320 days, at least 340 days, or at least 360 days.

[0425] (c) temperatures about 37°C for at least 10 days, such as at least 20 days, at least 30 days, at least 40 days, at least 50 days, at least 60 days, at least 70 days, at least 80 days, or at least 90 days, optionally wherein stability is determined by inspection of:

[0426] (i) sedimentation;

[0427] (ii) liquid phase separation;

[0428] (iii) discoloration; and / or

[0429] (iv) alteration of the sensory profile according to embodiment 36.

Claims

CLAIMS1 . A method for the production of a concentrated milk product having:(a) a milk protein content of at least 3.5% by weight; and / or(b) a total milk solids content of at least 15% by weight; wherein the method comprises the step of adding a transglutaminase enzyme to a precursor of the milk product to produce a bio-transformed precursor of the milk product.

2. The method of claim 1 , wherein the precursor comprises or consists of:(a) raw or untreated milk;(b) skimmed milk;(c) concentrated milk;(d) reconstituted milk powder;(e) evaporated milk;(f) reverse osmosis filtered milk; or(g) a mixture of any of (a) to (f).

3. The method of claim 1 or 2, wherein the precursor comprises:(a) a milk protein content of at least about 5% by weight; and / or(b) a total milk solids content about 20% by weight to about 25% by weight.

4. The method of any of claims 1 to 3, wherein the transglutaminase is added:(a) at about 1 .5% to about 2% by weight of the total protein content of the precursor; and / or(b) in an amount of about 140 U to 210 U per 100g of protein.

5. The method of any of claims 1 to 4, wherein the precursor is maintained at a temperature of about 55°C to about 60°C, subsequent to the addition of the transglutaminase.

6. The method of any of claims 1 to 5, wherein the step of transglutaminase addition is performed:(a) during wet mixing of ingredients;(b) subsequent to heat treatment and / or pasteurization of the precursor;(c) subsequent to homogenization of the precursor;(d) subsequent to standardization of the precursor; and / or(e) prior to sterilization.

7. The method of claim 6, wherein said heat treatment and / or pasteurization comprises heating the precursor to at least about 120°C, optionally wherein said temperature is maintained for at least 1 minute.

8. The method of any of claims 1 to 7, wherein the method additionally comprises the step of terminating, reducing, and / or inhibiting the activity of the transglutaminase, subsequent to the addition of the transglutaminase, optionally wherein said terminating, reducing, and / or inhibiting step comprises:(a) cooling the bio-transformed precursor and transglutaminase mixture to a temperature of less than about 10°C; and / or(b) heating the bio-transformed precursor and transglutaminase mixture to a temperature of at least about 75°C.

9. The method of any of claims 1 to 8, wherein the total volume of concentrated milk product, precursor and / or bio-transformed precursor is at least 100 liters.

10. The method of any of claims 1 to 9, wherein the method comprises the step of heat treatment and / or sterilization of the bio-transformed precursor by heating to at least about 120°C, optionally wherein the heat treatment and / or sterilization comprises or consists of:(a) retort sterilization;(b) direct steam injection sterilization; and / or(c) indirect sterilization.11 . The method of any of claims 1 to 10, wherein the concentrated milk product has:(a) a milk protein content of at least about 4% by weight; and / or(b) a total milk solids content of about 20% by weight to about 21% by weight, more preferably about 20.5%.

12. A concentrated milk product produced by the method of any of claims 1 to 11 .

13. A concentrated milk product, wherein the concentrated milk product has:(a) a milk protein content of at least 3.5% by weight; and / or(b) a total milk solids content of at least 15% by weight; and(c) cross-linked milk proteins; and(d) trace amounts of an inactivated or non-functional transglutaminase; and optionally(e) carrageenan.

14. The concentrated milk product of claim 12 or claim 13, wherein the concentrated milk product has:(a) a total milk solids content of about 20.5% by weight;(b) a milk lipid content of about 4% to about 5% by weight; and / or(c) a milk protein content of about 3.5% to about 6% by weight.

15. The concentrated milk product of any of claims 12 to 14, wherein the concentrated milk product is:(a) an evaporated milk product;(b) a condensed milk product; and / or(c) a creamer product; optionally, wherein the concentrated milk product:(i) has an improved sensory profile as compared with a concentrated milk product with substantially the same proportion of total milk solids that has not been subjected to transglutaminase treatment; and / or(ii) has a sensory profile that is substantially the same as a concentrated milk product with a higher proportion of total milk solids that has not been subjected to transglutaminase treatment;(iii) is stable at temperatures up to about 30°C for at least 270 days; and / or(iv) is stable temperatures between about 21°C and about 25°C for at least 270 days.(v) is stable at temperatures about 37°C for at least 90 days, optionally, wherein stability is determined by inspection of: sedimentation; liquid phase separation; discoloration; and / or alteration of the sensory profile.

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