Heat sterilized high protein enteral compositions with whey protein which comprises whey protein micelles and a source of casein

Abstract

The present invention is directed to a liquid heat sterilized enteral composition comprising: a protein source in an amount of 12 to 20% by weight of the composition, the protein source consisting of whey protein containing whey protein micelles and a source of casein. The inventive heat sterilized enteral composition preferably has a cysteine content of 1.2 to 2.4% by weight of the composition. The present invention is furthermore directed to a process for preparing a heat sterilized enteral composition comprising protein in an amount of 12 to 20% by weight based on the weight of the composition, said protein consisting of whey protein containing whey protein micelles and casein, which comprises the steps of: (i) Providing an aqueous solution of a protein source consisting of whey protein containing whey protein micelles and (ii) Adding a protein source containing casein; (iii) Optionally performing a homogenization treatment step; (iv) Performing a heat treatment step; (v) Optionally performing a homogenization treatment step. Finally, the invention concerns the use of a protein source consisting of whey protein containing whey protein micelles and a source of casein, for preparing an enteral composition and/or for controlling the viscosity of a liquid enteral composition, wherein the enteral composition comprises 12 to 20 weight % total protein. The invention also concerns medical uses and treatments applying or using the inventive heat sterilized enteral composition.

Claims

1. A liquid heat sterilized enteral composition comprising: a protein source in an amount of 12 to 20% by weight of the enteral composition, the protein source consisting of whey protein containing whey protein micelles, and a source of casein; wherein the weight ratio of the casein to the whey protein is in the range of 35:65 to 65:35; and wherein the enteral composition contains cysteine in an amount of 1.2 to 2.4% by weight of the protein source.

2. The enteral composition according to claim 1, wherein the whey protein is present in an amount of 4.5 to 9% by weight of the enteral composition.

3. The enteral composition according to claim 1, wherein the casein is present in an amount of 3.5 to 13% by weight of the enteral composition.

4. The enteral composition according to claim 1, wherein the enteral composition is a liquid composition having a viscosity below 200 mPa.Math.s at 20° C. measured at a shear rate of 100 s.sup.−1.

5. The enteral composition according to claim 1, having a protein concentration of up to 20 g/100 g of the enteral composition.

6. The enteral composition according to claim 1, which is in a form selected from the group consisting of a nutritional composition, a nutritional supplement, an infant formula, a follow-up formula, a baby food formula, an infant cereal formula or a growing-up milk, an infant or child's food supplement, a children formula, an adult nutritional composition, a maternal nutritional supplement, a bariatric formula, an elderly nutritional composition and a health care formula.

7. A liquid enteral composition comprising 12 to 20 weight % total protein based on the weight of the liquid enteral composition, the total protein consisting of whey protein containing whey protein micelles, and a source of casein for controlling the viscosity of the liquid enteral composition, wherein the weight ratio of the casein to the whey protein is in the range of 35:65 to 65:35, and wherein the liquid enteral composition has a pH of 6 to 8, and the liquid enteral composition contains cysteine in an amount of 1.2 to 2.4% by weight of the total protein.

8. A process for preparing a liquid heat sterilized enteral composition comprising total protein in an amount of 12 to 20% by weight of the liquid enteral composition, the total protein comprising casein and whey protein containing whey protein micelles, wherein the weight ratio of the casein to the whey protein is in the range of 35:65 to 65:35, the process comprising: (i) providing an aqueous solution of a protein source comprising whey protein containing whey protein micelles; (ii) adding a protein source containing casein; and (iii) performing a ultra-high temperature (UHT) heat treatment step above 140° C.

9. The process according to claim 8, wherein the whey protein is present in an amount of 4.5 to 9% by weight of the enteral composition.

10. The process according to claim 8, wherein the casein is present in an amount of 3.5 to 13% by weight of the enteral composition.

11. The process according to claim 8, wherein the liquid heat sterilized enteral composition has a viscosity below 200 mPa.Math.s at 20° C. measured at a shear rate of 100 s.sup.−1.

12. The process according to claim 8, further comprising drying to form a powder.

13. The process according to claim 8, wherein the total protein consists of casein and whey protein containing whey protein micelles.

14. The process according to claim 8, wherein the liquid enteral composition contains cysteine in an amount of 1.2 to 2.4% by weight of the total protein.

15. The process according to claim 8, wherein the UHT heat treatment is performed at 141-160° C.

16. The enteral composition according to claim 1, wherein the enteral composition contains cysteine in an amount of 1.8 to 2.2% by weight of the protein source.

17. The enteral composition according to claim 1, wherein the weight ratio of casein to whey protein is in the range of 45:55 to 55:45.

18. The enteral composition according to claim 1, wherein the whey protein micelles is present in an amount of 2.5 to 3.5% by weight of the enteral composition.

19. The enteral composition according to claim 7, wherein the liquid enteral composition has a pH of 6.7 to 6.9.

Description

FIGURES

(1) The following Figures are intended to illustrate the invention further. It is not intended to limit the subject matter of the invention thereto.

(2) FIG. 1: Shows a schematic structure for a whey protein micelle according to the present invention. The whey proteins are arranged in such a way that there are regions of hydrogen bonding and regions of hydrophobic bonding.

(3) Both physical and chemical interactions are involved in the whey protein micelle:

(4) s*: accessible thiol/activated thiol from cysteine.

(5) —S—S—: disulphide bridges stabilizing the whey protein micelle.

(6) FIG. 2: shows the microgram of the whey micelles dispersion, before the addition of other ingredients according to the inventive process, (A) Scale bar: 2 μm. (B) Scale bar: 1 μm.

(7) FIG. 3: shows the micrograph of the whey micelles dispersion, before the addition of the other ingredients (A), Example 1 (B), Example 2 (C) and Example 3 (D). In (B), (C) and (D) BLG proteins are localized by immunogold labelling (back dots). Scale bar: 1 μm.

(8) FIG. 4: Schematic for an exemplary inventive process. As one can see the inventive process may (i) provide an aqueous solution comprising whey protein containing whey protein micelles—by heating a whey protein solution. This may then be cooled and optionally fat added followed by mixing. Then (ii) the protein source containing casein is added along with other optional ingredients such as salts and CHO, which may then be (iii) optionally homogenized, then (iv) heat treated, then (v) optionally homogenized.

EXAMPLES

(9) The following examples are intended to illustrate the invention further. They are not intended to limit the subject matter of the invention thereto.

Example 1: Exemplary Process According to the Present Invention

(10) TABLE-US-00002 TS “as is” “dry” Ingredient Name [%] [kg] [kg] Water (Softened) — 68.07 — Milk protein concentrate, 95.2 10.20 9.71 85% protein content Whey protein isolate 94.6 6.52 6.17 Glucose Syrup (DE29) 95.6 7.50 7.17 Soybean Oil 100.0 5.00 5.00 Sucrose 99.9 2.50 2.50 Tripotassium Citrate monohydrate 0.15 0.14 Sodium Chloride 99.9 0.06 0.06 Total 100.00 30.7

(11) The inventive heat sterilized enteral composition according to the present invention containing the above ingredients was prepared as follows using the amounts as defined above. The demineralised water and whey protein isolate were mixed at 50° C. followed by hydration for 30 minutes. A pH adjustment to pH 6.2 was made with citric acid 5% followed by heating the mixture at 85° C. for 15 minutes to form the whey protein micelles, which was then cooled to 15° C.

(12) Soybean oil was then added followed by mixing at 15° C., after which Glucose Syrup (DE29), tripotassium citrate monohydrate, sucrose, sodium chloride and the milk protein concentrate, 85% protein content (casein source) were added with mixing and hydration for 40 minutes.

(13) The mixture was then pre-heated to 50° C. followed by homogenization at 250+50 bar, preheated to 80° C. followed by a UHT treatment by direct steam injection at 148° C. for 5 seconds. This was then flash cooled to 78° C. and the mixture homogenized at 200+50 bar.

(14) Thus, as described above the heat sterilized enteral composition according to the present invention was prepared which contained 14.1% by weight protein, with the following profile:

(15) TABLE-US-00003 (m)g/100 (m)g/100 mg/100 g mL Kcal Protein g/100 g or mL 14.1 15.3 Fat g/100 g or mL 4.9 5.3 Calcium mg/100 g or mL or Kcal 252 274 172 Magnesium mg/100 g or mL or 14.6 16 10 Kcal Sodium mg/100 g or mL or Kcal 40 43 27 Potassium mg/100 g or mL or Kcal 121 132 82 Phosphorus mg/100 g or mL or 155 168 106 Kcal Chloride mg/100 g or mL or Kcal 47 51 32 Citrates mg/100 g or mL or Kcal 190 206 129 21.3 Carbohydrates (difference) 12.7 Kcal/100 mL (calculated) 160 Notes: Na.sup.+ + K.sup.+ mg/g protein = 11.4 pH at 25° C. = 6.7 Density @ 20° C. g/cm3 = 1.0868 Total solids = 31.3 g/100 g

(16) Furthermore, the amino acids profile was as follows:

(17) TABLE-US-00004 Amino Acid mg/100 g Tryptophane 0.250 Methionine, L 0.373 Cystine, L 0.242 Aspartic acid 1.360 Threonine 0.693 Serine 0.781 Glutamic acid 3.000 Proline 1.190 Glycine 0.283 Tyrosine 0.644 Phenylalanine 0.645 Histidine 0.348 Lysine 1.300 Alanine 0.577 Valine 0.858 Isoleucine 0.768 Leucine 1.570 Arginine 0.456 Notes: Cystine + Methionine Total [g/100g] = 0.615 Methionine/Cystine (Ratio) = 1.54

(18) Ratio whey/casein calculated based on the equation reported in Ballin, 2006: 48/62
% whey protein=(C−0.25)/(3.0−0.25)×100=48%
C=percentage of cystine on 100% protein=1.58%

(19) Said heat sterilized liquid enteral composition was without any taste/bitterness. Furthermore this had a viscosity of 44 mPa.Math.s at 20° C./100 s.sup.−1 as determined using a rheometer (Haake Rheostress 6000 coupled with UMTC) equipped with a plate/plate geometry (60 mm diameter) and 1 mm gap. Flow curves 0-300 s.sup.−1 (linear increase) were obtained at controlled shear rate at 20° C.+/−0.1.

(20) Furthermore, the obtained liquid enteral composition was shelf stable for 1 year.

Example 2: Exemplary Process According to the Present Invention

(21) TABLE-US-00005 TS “as is” “dry” Ingredient name [%] [kg] [kg] Water (Softened) — 55.03 — Milk protein concentrate, 95.2 9.00 8.57 85% protein content Whey protein isolate 94.6 6.52 6.17 Glucose Syrup (DE29) 95.6 12.70 12.14 Soybean Oil 100.0 8.40 8.40 Sucrose 99.9 8.20 8.19 Tripotassium Citrate 0.15 0.14 Total 100.00 43.6

(22) The inventive heat sterilized enteral composition according to the present invention containing the above ingredients was prepared as follows using the amounts as defined above. The demineralised water and whey protein isolate were mixed at 50° C. followed by hydration for 30 minutes. A pH adjustment to pH 6.2 was made with citric acid 5% followed by heating the mixture at 85° C. for 15 minutes to form the whey protein micelles, which was then cooled to 15° C.

(23) Soybean oil was then added followed by mixing at 15° C., after which Glucose Syrup (DE29), tripotassium citrate monohydrate, sucrose and the milk protein concentrate, 85% protein content (casein source) were added with mixing and hydration for 40 minutes.

(24) The mixture was then preheated to 80° C. followed by a UHT treatment by direct steam injection at 148° C. for 5 seconds. This was then flash cooled to 78° C. and the mixture homogenized at 200+50 bar.

(25) Thus, a heat sterilized enteral composition according to the present invention was prepared which contained 14.2% by weight protein with the following profile:

(26) TABLE-US-00006 (m)g/100 (m)g/100 mg/100 g mL Kcal Protein g/100 g or mL 14.2 15.9 Fat g/ 100g or mL 7.3 8.2 Calcium mg/100 g or mL 252 282 127 Magnesium mg/100 g or mL 14.4 16 7 Sodium mg/100 g or mL 20 22 10 Potassium mg/100 g or mL 115 129 57 Phosphorus mg/100 g or mL 155 173 78 Chloride mg/100 g or mL 17 19 9 Citrate mg/100 g or mL 190 212 95 24.7 Carbohydrates (difference) 21.7 Kcal/100 mL (calculated) 224 Notes: Na.sup.+ + K.sup.+ mg/g protein = 9.5 pH at 25° C. = 6.8 Density @ 20° C. g/cm3 = 1.1184 Total solid = 41.5 g/100 g

(27) Furthermore, the amino acids profile was as follows:

(28) TABLE-US-00007 Amino Acid [g/100 g] Tryptophane 0.242 Methionine, L 0.374 Cystine, L 0.240 Aspartic acid 1.370 Threonine 0.701 Serine 0.790 Glutamic acid 3.050 Proline 1.220 Glycine 0.291 Tyrosine 0.642 Phenylalanine 0.653 Histidine 0.360 Lysine 1.320 Alanine 0.592 Valine 0.875 Isoleucine 0.779 Leucine 1.600 Arginine 0.459 Cystine + Methionine Total [g/100g] = 0.614 Methionine/Cystine (Ratio) = 1.56.

(29) Ratio whey/casein calculated based on the equation reported in Ballin, 2006: 47/53
% whey protein=(C−0.25)/(3.0−0.25)×100=47%
C=percentage of cystine on 100% protein=1.54%

(30) Said heat sterilized liquid enteral composition was without any taste/bitterness. Furthermore this had a viscosity of 116 mPa.Math.s at 20° C./100 s.sup.−1 as determined using a rheometer (Haake Rheostress 6000 coupled with UMTC) equipped with a plate/plate geometry (60 mm diameter) and 1 mm gap. Flow curves 0-300 s.sup.−1 (linear increase) were obtained at controlled shear rate at 20° C.+/−0.1.

(31) Furthermore, the obtained liquid enteral composition was shelf stable for 1 year.

Example 3: Exemplary Process According to the Present Invention

(32) TABLE-US-00008 TS “as is” “dry” Ingredient name [%] [kg] [kg] Water (Softened) — 57.53 — Milk Protein concentrate, 95.2 10.20 9.71 85% protein content Whey protein isolate 94.6 6.52 6.17 Glucose Syrup (DE29) 95.6 10.80 10.32 Soybean Oil 100.0 7.90 7.90 Sucrose 99.9 6.90 6.89 Tripotassium Citrate 0.15 0.14 Total 100.00 41.1

(33) The inventive heat sterilized enteral composition according to the present invention containing the above ingredients was prepared as follows using the amounts as defined above. The demineralised water and whey protein isolate were mixed at 50° C. followed by hydration for 30 minutes. A pH adjustment to pH 6.2 was made with citric acid 5% followed by heating the mixture at 85° C. for 15 minutes to form the whey protein micelles, which was then cooled to 15° C.

(34) Soybean oil (was then added followed by mixing at 15° C., after which Glucose Syrup (DE29), tripotassium citrate monohydrate, sucrose and the milk protein concentrate, 85% protein content (casein source) were added with mixing and hydration for 40 minutes.

(35) The mixture was then pre-heated to 50° C. followed by homogenization at 250+50 bar, preheated to 80° C. followed by a UHT treatment by direct steam injection at 148° C. for 5 seconds. This was then flash cooled to 78° C. and the mixture homogenized at 200+50 bar.

(36) Thus, as described above the heat sterilized enteral composition according to the present invention was prepared which contained 12.9% by weight protein, with the following profile:

(37) TABLE-US-00009 (m)g/100 (m)g/100 mg/100 g mL Kcal Protein g/100 g or mL 12.9 14.6 Fat g/100 g or mL 7.7 8.7 Calcium mg/100 g or mL 223 251 106 Magnesium mg/100 g or mL 13.1 15 6 Sodium mg/100 g or mL 19 21 9 Potassium mg/100 g or mL 118 133 56 Phosphorus mg/100 g or mL 136 153 65 Chloride mg/100 g or mL 17 19 8 Citrate mg/100 g or mL 180 203 85 23.9 Carbohydrates (difference) 25.3 Kcal/100 mL (calculated) 238 Notes: Na.sup.+ + K.sup.+ mg/g protein = 10.6 pH at 25° C. = 6.7 Density @ 20° C. g/cm3 = 1.1268 Total solid = 43.6 g/100 g

(38) Furthermore, the amino acids profile was as follows:

(39) TABLE-US-00010 Amino Acid [g/100 g] Tryptophane 0.231 Methionine, L 0.346 Cystine, L 0.227 Aspartic acid 1.240 Threonine 0.629 Serine 0.694 Glutamic acid 2.710 Proline 1.060 Glycine 0.256 Tyrosine 0.557 Phenylalanine 0.578 Histidine 0.315 Lysine 1.160 Alanine 0.531 Valine 0.774 Isoleucine 0.695 Leucine 1.430 Arginine 0.402 Cystine + Methionine Total [g/100 g] = 0.573 Methionine/Cystine (Ratio) = 1.52.

(40) Ratio whey/casein calculated based on the equation reported in Ballin, 2006: 51/49
% whey protein=(C−0.25)/(3.0−0.25)×100=51%
C=percentage of cystine on 100% protein=1.64%

(41) Said heat sterilized liquid enteral composition was without any taste/bitterness. Furthermore this had a viscosity of 125 mPa.Math.s at 20° C./100 s.sup.−1 as determined using a rheometer (Haake Rheostress 6000 coupled with UMTC) equipped with a plate/plate geometry (60 mm diameter) and 1 mm gap. Flow curves 0-300 s.sup.−1 (linear increase) were obtained at controlled shear rate at 20° C.+/−0.1.

(42) Furthermore, the obtained liquid enteral composition was shelf stable for 1 year.

Example 4: Transmission Electron Microscopy

(43) Protocol:

(44) Samples prepared according to examples 1, 2 and 3 above (termed also “recipes 1, 2 and 3”) are fixed in 3.7% paraformaldehyde in PBS buffer (pH: 7.3) after their reception. Fixed samples are embedded in 4% aqueous agar solution and solidified on ice. Small cubes of 1 mm.sup.3 are cut and then the samples are dehydrated in graded series of ethanol solution, from 30% to 100%, 30 minutes each bath, and 3 times 1 hour for the 100% ethanol solution. Samples are then gradually infiltrated in graded LR White resin series prior to final infil-tration in pure resin 3 times for 1 hour each. The polymerization is carried out at 60° C. for 48 hours. After polymerization, ultrathin sections of 70 nm are sliced and collected on 100 mesh nickel grids.

(45) For BLG-immunolabelling, grids are placed on drops of 2.5% BSA (w/v) solution in Tris pH 7.4 for 15 min. Grids are then transferred onto drops of Tris containing 1/200 anti-BLG for 1 night at 4° C. They are rinsed 3 times with Tris before incubation on drops of Protein A gold 15 nm diluted 1/30 for 30 min. Then, grids are rinsed with 3 times with Tris before 3 final rinsing steps with water.

(46) Samples are imaged with a Tecnai Spirit BioTWIN using a LaB6 filament at 80 kV electron microscope (FEI, Netherlands).

(47) The results are shown in FIG. 3. FIG. 3 shows the micrograph of the whey micelles dispersion, before the addition of the other ingredients (A), and the final heat sterilized enteral composition according to Example 1 (B), Example 2 (C) and Example 3 (D) herein before.

(48) In all 3 samples (FIG. 3 B, C and D), many protein aggregates (arrowheads) were observed. They are usually in close contact with fat globules, which are, in general, very small. Proteins aggregates are only partially spherical and are in particular, but not only, deformed when stabilizing fat globules (FIG. 3 b, C and D, arrows). The presence of BLG is shown by immunogold labelling in the aggregates. It does not reveal the presence of BLG outside the aggregates. These data clearly evidence that whey protein micelles were contained in the final heat sterilized enteral compositions and were detectable.