JPH08509366A - Protein hydrolysis method - Google Patents

Abstract

  (57) [Summary] A protein derived from Aspergillus oryzae and comprising at least 5 proteolytic components each having a molecular weight of about 23 kD, 27 kD, 31 kD, 32 kD, 35 kD, 38 kD, 42 kD, 47 kD, 53 kD and 100 kD. Hydrolysis of vegetable or animal proteins by incubating with degradative enzyme preparations, such as Flavorzyme ™, is a process for fermentation of food products such as milk substitutes, cheese, HVP, meat extracts, flavoring agents and food products. Provide protein hydrolysates useful in or as auxiliaries or non-food products such as pet food, makeup. This method results in a high degree of hydrolysis (DH), flavor development and high protein solubility.

Description

TECHNICAL FIELD The present invention relates to a protein hydrolysis method, a protein hydrolyzate obtained by the method, and a product other than a food containing the protein hydrolyzate. Background Art Protein hydrolysis methods have been described, for example, in British Patent 1,507,380, US Patent 3,723,250 and International Publications WO 89/00272, WO 92/13964 and WO 90/05462. There is a need for methods of hydrolyzing proteins that result in hydrolysates that have a high degree of protein hydrolysis and excellent organoleptic properties. SUMMARY OF THE INVENTION The following has now been found: a proteolytic enzyme preparation derived from Aspergillus oryzae and commercially available from Novo Nordisk A / S under the trademark "Flavourzyme". The product has excellent proteolytic properties, for example it is possible to obtain a high degree of hydrolysis and a bitter-free hydrolyzate. The present invention therefore relates to a process for the hydrolysis of proteins by incubation with a proteolytic preparation derived from Aspergillus oryzae and marketed by Novo Nordisk A / S under the trademark "Flavourzyme". In its second aspect, the present invention provides a protein hydrolyzate obtained by the method of the present invention. In another aspect, it relates to food products and non-food products comprising the protein hydrolyzate of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS The present invention will be further described with reference to the accompanying drawings. FIG. 1 shows the time (hours) of hydrolysis versus the time of hydrolysis achieved by the method of the present invention applied to sodium casein within the pH range of pH 5-9 (■: pH 5, ▲ pH 6, ○ pH 7, □ pH 8, ● pH 9). Fig. 2 shows the degree of hydrolysis (% DH); Fig. 2 shows the degree of hydrolysis (% DH) of the soybean protein isolate after 22 hours of hydrolysis according to the method of the present invention (■: flavorzyme, ▲: corolase (Corolase). ) (Trademark) 7092, ◇: Corolase (trademark) 7093, ◆: Alcalase (trademark), □: Neutrase (trademark); and FIG. After the decomposition, the degree of hydrolysis (% DH) of Na-caseinate is shown (■: Flavorzyme (trademark), ▲: Corolase (trademark) 7092, ◇: Corolase (trademark) 7093, ◆: Alcalase (trademark), □ : Neutrase ( DETAILED DESCRIPTION OF THE INVENTION The characterization of Flavorzyme ™ has shown that the proteolytic Aspergillus oryzae preparation comprises several proteolytic components. Probably containing one or more proteolytic enzyme components, each of which has an approximate molecular weight of 23kD, 27kD, 31kD, 32kD, 35kD, 38kD, 42KD, 47KD, 53KD and 100KD. Therefore, the present invention is derived from Aspergillus oryzae and has at least 5 proteolytic cleavages with an approximate molecular weight selected from 23 kD, 27 kD, 31 kD, 32 kD, 35 kD, 38 kD, 42 kD, 47 kD, 53 kD and 100 kD, respectively. A method for hydrolyzing a protein by incubating with a proteolytic enzyme preparation comprising the components is provided. In one embodiment, the protein is derived from Aspergillus oryzae and incubated with a proteolytic preparation comprising at least 5 proteolytic preparations having approximate molecular weights of 23 kD, 31 kD, 35 kD, 38 kD and 53 kD, respectively. The molecular weight of the protease component in the product was measured by SDS polyacrylamide gel electrophoresis (SDS-PAGE) by a method known to those skilled in the art, and the molecular weight of each protease component was measured by this method. The method of the invention is capable of performing a wide range of protein hydrolysis of proteins, and this method results in a hydrolyzate which is bitter-free as well as having a pronounced soup / meat taste. The degree of protein hydrolysis can be determined by the degree of hydrolysis achieved. In the context of the present invention, the degree of hydrolysis (DH) is defined by the formula: DH can be calculated according to Adler Nissen; Enzymic Hydrolysis of Food Proteins; Els Brewer Applied Science Publishing Co. (1986), page 122. By using the method of the present invention, it is possible to obtain DH of 35% or more, preferably 60% or more, more preferably 70% or more, and most preferably 80% or more. It is also possible to obtain a high degree of protein solubility by the method of the present invention. The degree of protein solubility can be described by the protein solubility index (PSI) as described by Adler Nissen (supra). In a preferred embodiment, a protein solubility of 50% PSI or higher, preferably 70% PSI or higher, more preferably 90% or higher is obtained. The protein or proteinaceous substance which can be advantageously hydrolyzed by the method according to the invention may be any vegetable protein, such as soy protein, grain protein such as wheat gluten or zein, rapeseed protein, purple horse mackerel protein, noodles. Any protein, legume broad bean protein, cottonseed protein or sesame seed protein, or any animal protein or proteinaceous substance, such as milk protein, whey protein, casein, meat protein, fish protein, blood It may be protein, egg white or gelatin. In order to obtain a satisfactory degree of hydrolysis, the proteolytic enzyme is preferably added to the protein or protein-like substance in an amount of 0.05-5 AU per 100 g of protein, in particular 0.1-8 AU per 100 g of protein. Incubation can be performed at a pH of about 4 to about 10, preferably about 5 to about 9. As shown in Example 2, the present invention can be performed excellently even under extreme pH conditions, i.e. pH values in all in the range 5-9. Incubation can be performed at any convenient temperature within which the enzyme preparation is not inactivated, ie, in the range of about 20 ° C to about 70 ° C. According to established practice, the proteolytic enzyme preparation may be suitably inactivated by increasing the temperature of the incubation mixture above about 70 ° C or by reducing the pH of the incubation mixture below about 4.0. In another preferred embodiment, the incubation of the protein or protein-like substrate can be carried out with a combination of Flavorzyme ™ and one or more other protease preparations. A preferred protease preparation comprises neutral or alkaline proteases. Examples of suitable neutral proteases are neutral proteases from Bacillus, preferably neutral proteases from Bacillus subtilis, such as the enzyme preparations marketed by Novo Nordisk (Denmark) under the trade name Neutrase. Is. Examples of suitable alkaline proteases are those from Bacillus, preferably those from Bacillus licheniformis, such as those commercially available from Novo Nordisk (Denmark) under the trade name Alcalase and as an active ingredient subtilisin A. (Subtilisin carlsberg) is an enzyme preparation containing the same. The incubations carried out in the method of the present invention can also be carried out using a combination of Flavourzyme ™ and one or more other lipase preparations. A preferred lipase preparation comprises fungal lipase. Examples of suitable fungal lipases are lipases from Mucor, preferably lipases from Rhizomucor miehei, such as the enzyme preparation commercially available from Novo Nordisk (Denmark) under the trade name Palatase. A product; and a lipase derived from Aspergillus, preferably a lipase derived from Aspergillus niger, eg an enzyme preparation marketed by Novo Nordisk (Denmark) under the trademark Palatase A . In another aspect, the invention provides a protein hydrolyzate obtained by the method of the invention. Measurement of AU Proteolytic activity can be measured using hemoglobin as a substrate. In the Anson-hemoglobin method for measuring proteolytic activity, denatured hemoglobin is digested and undigested hemoglobin is precipitated with trichloroacetic acid (TCA). The amount of TCA soluble product is measured with a phenolic reagent which gives a blue color with respect to tyrosine and tryptophan. One Anson unit (AU) is a TCA soluble product that gives the same color as phenol equivalent to 1 milliequivalent of tyrosine per minute at standard rate under standard conditions (ie 25 ° C, pH 7.5 and reaction time of 10 minutes). It is defined as the amount of enzyme that liberates the amount of material. Folded print AF 4/5, which describes a more detailed analysis method, is available on request from Novo Nordisk A / S, DK-2880, Bagsbaert, Denmark, which print is referred to herein by its number. Included in the calligraphy. Measurement of LU Analysis of lipase activity: A substrate for lipase was prepared by emulsifying glycerin-tributyrate (MERCK) using gum arabic as emulsifier. Lipase activity was analyzed at pH 7 using the pH stat method. One unit of lipase activity (LU / mg) is defined as the amount required to release 1 micromol of fatty acid per minute. Step 1: Centrifuge the fermentation supernatant and drain the precipitate. The pH of the supernatant is adjusted to 7 and then an equal volume of cold 96% ethanol is added slowly. The mixture is left in the ice bath for 30 minutes. Centrifuge then drain the precipitate. Step 2: Ion exchange chromatography. The supernatant is filtered and then applied to a DEAE-fast flow (Pharmacia ™) column equilibrated with 50 mM Tris-acetate buffer (pH 7). The column is washed with the same buffer until the absorption at 280 nm is less than 0.05 OD. The bound enzyme activity is eluted with a linear salt gradient of the same buffer (0 to 0.5 M NaCl) using 5 column volumes. Step 3: Hydrophobic chromatography. The molarity of the pool containing enzyme activity is adjusted to 0.8 M by adding solid ammonium acetate. The enzyme is applied to a TSK gel butyl-Toyopearl 650C column (available from Tosoh Corporation, Japan) that has been pre-equilibrated with 0.8 M ammonium acetate. Unbound material is washed with 0.8 M ammonium acetate and the bound material is eluted with distilled water. Step 4: Dilute the pool containing lipase activity with water to adjust the conductance to 2 mS and pH. The pool is applied to a fast Q Sepharose (Pharmacia) column pre-equilibrated with 50 mM Tris-acetate buffer (pH 7). Bound enzyme is eluted with a linear salt gradient. Food Product In another aspect, the invention relates to a food product comprising the protein hydrolyzate of the invention. The amount of protein hydrolyzate incorporated into the food product will typically be in the range 1-30% by weight. An important food product of the invention is, for example, a component of breast milk substitutes. Due to the high degree of hydrolysis obtained by the method of the present invention, the protein hydrolyzate of the present invention can be conveniently incorporated into a milk substitute, which hydrolyzate has significantly lower allergenicity than unhydrolyzed milk protein. Have. The milk replacer can be formulated in substantially the same manner as shown in the prior art literature for this type of product (eg European patent application 322,589), except that the protein hydrolysates contained in known products are Is replaced by the protein hydrolyzate of the present invention. The food product of the invention may also contain a protein hydrolyzate of the invention as a protein enhancer or to provide other properties of the food product. Therefore, the protein hydrolyzate that is incorporated into the food product is meat that has been torn from the bone by subjecting the crushed bone to the method of the present invention or cut meat (e.g. so-called mechanically recovered meat, That is, it can be based, for example, on the meat that remains on the bone after a normal piece of meat has been carved from an animal carcass in a slaughterhouse. For a more detailed description of the general procedure, see Applicant's co-pending International Patent Application WO 90/05462. Other protein by-products from the meat industry can also be used. The resulting protein hydrolyzate can then be suitably added to emulsified meat products such as sausages or pates, or alternatively as a seasoning ingredient in soups or other food products. The food product obtained from bovine milk by the method of the present invention is a cheese flavored product. Extensive hydrolysis of milk proteins results in flavor compounds with a very distinct cheese flavor. Such cheese flavored products of the present invention find preferred application in snack products, simulated cheese products, or generally as cheese flavor enhancers. Cheese flavor can be adjusted by the use of lipase according to well-established practices. The traditional method for producing hydrolyzed vegetable protein (HVP) for use as a seasoning product is based on boiling at high temperature and for a long time in hydrochloric acid. This is known to cause the undesired formation of chlorine containing compounds. By the method of the present invention it is now possible to obtain enzymatically produced HVP products, which are considered to be healthier products. The high DH obtainable by the method of the present invention provides desirable flavor and flavor enhancing properties. The method of the present invention can also be used for dietary products that are rich in protein due to the bitter-free flavor obtained by the method of the present invention. The extremely high protein solubility in the very wide pH range relevant to the method of the invention is also advantageous. Surprisingly the following has been found: a good fermentation medium can be produced by subjecting milk to the process of the invention. Such milk hydrolysates can be used by adding in relatively small amounts to enhance fermentation in yogurt production or in the production of lactic starter bacterial cultures. Shortening the process time increases the production capacity and reduces the risk of infection, eg bacteriophage infection. Therefore, the method of the invention can be used in combination with a fermentation process, preferably a fermentation process for the production of food products. Thus, the proteolytic preparations used in the method of the invention, such as Flavorzyme ™ or the protein hydrolysates of the invention or both, enhance the productivity of the fermentation process in a fermentation process, preferably a food fermentation process. Can be added for. Examples of such fermentation processes include or are fish (fish sauce), cocoa beans or soybeans (eg soy sauce, tempeh, miso) or are fermentation processes. The addition of proteolytic enzymes or protein hydrolysates is useful for reducing the total process time, ie increasing the fermentation rate. Non-food products The method of the invention may find its application in the non-food field. The flavor profile obtained according to the invention is advantageously used for the production of pet food. The extremely high production of DH-hydrolyzate from gelatin improves gelatin products to be incorporated into cosmetics such as creams and shampoos. It is expected that the above-mentioned special effects as a fermentation medium will make the hydrolyzate of the present invention useful for other fermentations as well. The invention is further illustrated by the following examples, which in no way limit the scope of the invention claimed. Example 1 Beef Hydrolysis In this example the following enzyme preparations (expressed in% w / w on protein content basis) were used: Preparation A) 2% Flavorzyme ™ (2.30AU) / G), Preparation B) 1% Flavorzyme ™, 2% Neutrase ™ 0.5L, and 0.15% Alcalase ™ 2.4L. All enzymes are available from Novo Nordisk A / S, Denmark. Flavorzyme ™ is a proteolytic preparation derived from Aspergillus oryzae, Neutrase ™ 0.5L is a proteolytic preparation derived from Bacillus subtilis. And Alcalase 2.4L is a proteolytic preparation from Bacillus licheniformis. 408 g of beef was chopped twice and mixed with 392 g of water to give a protein content of 10% w / w. The mixture was mixed for 30 seconds and then heated to 55 ° C. Initial osmolality (mOsm) and soluble dry matter (° Brix) were measured and subsequently hydrolysis was monitored by these values. After incubation for 4 hours, the enzyme was inactivated by heating at 90 ° C for 30 minutes. The hydrolyzate was subjected to centrifugation at 3000 x G for 15 minutes and diluted. The centrifuge was stored in the refrigerator until the next day, the lipid phase separated and weighed. The pH of the centrifuge was 5.85. The results are shown in Tables 1 and 2 below. As calculated from the table, the dose of Flavorzyme ™ used in these experiments typically increased the yield from 45% to 81.5%, and the solubility usually from 57% to 89.9% PSI. To do. A degree of hydrolysis of 70.2% DH was achieved. The hydrolyzate was stored in a 4% solution before the test panel. The meat flavor was most pronounced in the hydrolyzate obtained with Preparation A. Example 2 Hydrolysis at Different pHs In this experiment, the process according to the invention was carried out with sodium caseinate as substrate at pH values in the range 5-9. An 8% (w / w) solution of sodium caseinate (Miprodan ™ 30, obtained from MD Hood Amba (Denmark)) was prepared by heating to 80 ° C. To 800 g of this solution was added 1 g of methyl-4-hydroxybenzoate and 0.15 g of propyl-4-hydroxybenzoate, then the temperature of the solution was reduced to 50 ° C. The pH was adjusted to the desired value with 4N NaOH or 4N HCl. Flavorzyme ™ (2.30 AU / g, Novo Nordisk A / S, Denmark) at 1% w / w based on protein content was added to the solution followed by incubation to allow maximum hydrolysis. (About 22 hours). At pH 9.00, 8.00, and 7.00, hydrolysis was performed in a pH stat, then incubation was monitored by NaOH consumption. Hydrolysis was monitored by decreasing pH and increasing osmolality at pH 6.00 and 5.00 (initial pH). Samples were obtained after 0.25, 1, 2, 4.5 and 22 hours of incubation. The enzyme was inactivated by heating at 85 ° C for 3 hours and then cooling in ice-water. The results are shown in Fig. 1. As is clear from this figure, the method of the present invention can give hydrolysis in the pH range of pH 5 to pH 9. Example 3 Comparative Example In this example, the degree of hydrolysis (DH) obtained by the method of the invention (incubation with Flavorzyme ™) is incubated with other known proteolytic preparations. It was compared with the DH obtained by Starting at pH 7.00 adjusted with 4N NaOH, experiments were performed as non-pH stat hydrolysis (no pH adjustment during hydrolysis) at 50 ° C. Each hydrolysis was performed with the same proteolytic activity expressed in AU / g (Note: see above for definition of AU) to ensure the same enzyme dose. Two protein sources were tested: 1) Na-caseinate (commercially available from MD Foods Amba (Denmark)) 2) Soy protein isolate (commercially available from Protein Technology International (USA)). 2500 g of 8% (w / w based on protein content) solution of protein raw material was prepared. The protein solution was heat treated at 85 ° C for 3 minutes and then cooled to a hydrolysis temperature of 50 ° C. A 400 g sample was prepared for hydrolysis. The following enzymes and enzyme doses were used: The hydrolysis was continued for 22 hours. The extent of hydrolysis was measured 5 and 22 hours after hydrolysis, respectively. The results are shown in Figure 2 (soy protein isolate) and Figure 3 (Na-caseinate). The results show that hydrolysis with Flavorzyme produces the highest degree of hydrolysis. Example 4 Cheese flavored whole milk was heat treated at 100 ° C for 2 minutes and then cooled to 50 ° C. Four portions of 200 ml each were prepared (% w / w based on protein content) 1) 1% Flavorzyme ™ (2.30 AU / g) 2) 1% flavorzyme ( Flavourzyme (TM) (2.30 AU / g) + 0.15% Palatase (TM) M 200L (Novo Nordisk A / S, commercially available from Denmark 200 LU / g) 3) 0.15% Palatase (TM) ) M 200 L (Novo Nordisk A / S, commercially available from Denmark 200 LU / g) 4) Control Hydrolysis was carried out at 50 ° C. for 20 hours. A strong cheese flavor appeared in Samples 1, 2 and 3, while the control was mostly neutral. Flavor-Samples 1, 2 and 3 are now diluted in cream cheese and a fourth sample was prepared as a control. 1.1) 0.5g sample No. 1 + 20g cheese 2.1) 6.0g sample No. 2 + 20g cheese 3.1) 6.0g sample No. 3 + 20g cheese 4.1) 6.0g fresh milk + 20g cheese. Sample No. 1.1 (Flavourzyme ™) appeared to have a very strong and intense flavor and a taste of ripe cheese. Sample No. 2.1 (Flavourzyme ™ + Paratase ™) appeared to have a well balanced cheese flavor. Compared to samples No.1.1 and No.2.1, sample No.3.1 gave the leanest taste impression. The reference sample No.4.1 was evaluated to be sour / slightly sour. In conclusion, it is possible to add the enzyme Flavorzyme ™ and the combination of Flavorzyme ™ and Palatase ™ directly to the milk to form an excellent cheese flavor. The traditional method of producing cheese flavor is performed by adding lipase and protease to cheese. Thus, flavorzymes in combination with Flavorzyme ™ as well as Palatase ™ provide a new and much simpler way to produce cheese flavor. Example 5 Accelerated yogurt fermented whole milk was heat treated at 100 ° C for 2 minutes and subsequently cooled to 50 ° C. Two 200 ml portions were prepared (% w / w based on protein content): 1) 1% Flavorzyme ™ (2.30 AU / g) 2) Control Hydrolysis was carried out at 50 ° C. for 20 hours. The enzyme was then denatured by heat treatment at 80 ° C for 5 minutes. Skim milk was heat treated at 90 ° C. for 2 minutes and then 200 ml of these samples were prepared. 1.1) 200 ml milk + 10 ml sample No.1 2.1) 200 ml milk + 10 ml sample No.1 3.1) 200 ml milk Adjust the temperature to 41 ° C and then Chr. Yogurt culture YC DVS from Hansense Laboratory was added to all three samples at a level of 10 ⁶ bacteria per gram. The fermentation curve was followed by measuring pH, see Table 4 below. From the pH measurement it is clear that: the addition of Flavorzyme ™ hydrolyzed milk promotes fermentation, which means that the production time of fermented dairy products can be significantly reduced. .

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI // (C12N 9/20 C12R 1: 685) (C12N 9/20 C12R 1: 785) (C12N 9/54 C12R 1:10) (C12N 9/54 C12R 1: 125) (C12N 9/62 C12R 1: 685) (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FR, GB, GR, IE) , IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AU, BB, BG, BR, BY, CA, CN, CZ, FI, HU, JP, KP, KR, KZ, LK, LV, MG, MN, MW, NO, NZ, PL, RO, RU, SD, SK, UA ， S, UZ, VN (72) inventor Hansen, Kim Denmark, Deko -4632 Beberusu Kou, Kasutanyiebai 30 (72) inventor Budorufusen, Gitte Denmark, Deko -2000 Furederi Kkusuberuu, 3. tuberculosis. Liebye 3

Claims (1)

[Claims]   1. Protein by incubating with proteolytic enzyme preparation A method of hydrolyzing proteins, in which the proteolytic preparation is Aspergillus oryzae ( Aspergillus oryzae) and each 23kD, 27kD, 31kD, 32kD respectively Approximate molecule selected from 35kD, 38kD, 42kD, 47kD, 53kD, and 100kD Such a method comprising at least 5 proteolytic components in an amount.   2. Protein preparations have approximate molecular weights of 23kD, 31kD, 35kD, 38kD And at least 5 proteolytic components having 53 kD. The method according to claim 1.   Tamper of more than 3.35%, preferably more than 60%, more preferably more than 70%, especially more than 80% 3. The method according to claim 1 or 2, wherein the degree of hydrolysis (DH) is obtained.   4. Over 50% PSI, preferably over 70% PSI, more preferably over 90% PSI The method according to claim 1, wherein a protein solubility index (PSI) is obtained.   5. The protein is a vegetable protein, preferably soy protein; grain tamper Quality, such as wheat gluten or zein; rapeseed protein; alpha protein Quality; Pea Protein; Legume Broad Bean Protein; Cottonseed Protein; or Go The method according to any one of claims 1 to 4, which is a real protein.   6. The protein is an animal protein, preferably milk protein, whey tongue Protein, casein, meat protein, fish protein, blood protein, egg white or zebra The method according to any one of claims 1 to 4, which is ratin.   7. Incubation is at a pH of about 4 to about 10, preferably about 5 7. A method according to any one of claims 1-6, carried out at a pH of about 9.   8. Tampa combining protein with one or more protease preparations 8. Incubating with a quality degradation preparation, any one of claims 1-7. The method described in the section.   9. The protease preparation is Bacillus, preferably Bacillus A neutral protease derived from Bacillus subtilis. The method according to item 8.   Ten. The protease preparation is Bacillus, preferably Bacillus It is an alkaline protease derived from Keniformis (Bacillus licheniformis), The method according to claim 8.   11. Proteins that combine proteins with one or more lipase preparations 11. Incubation with a quality-prepared preparation, according to any one of claims 1-10. The method described in.   12. The lipase preparation is Mucor, preferably Rhizomucor Claims comprising a fungal lipase derived from Rhizomucor miehei The method described in paragraph 11.   13. The lipase preparation is Aspergillus, preferably Aspergillus. A contract comprising a fungal lipase from Aspergillus niger. The method described in the scope of claim 11.   14. Claims where the protein is hydrolyzed and simultaneously fermented into a food product. The method according to any one of items 1 to 13.   15． A tan obtained by the method according to any one of claims 1 to 13. Pak hydrolyzate.   16. Hydrolyzed protein is milk protein, whey protein, casein, A group consisting of meat protein, fish protein, blood protein, egg white or gelatin Is an animal protein selected from A protein hydrolyzate according to claim 15.   17． Hydrolyzed protein is soybean protein; grain protein such as wheat flour Ruten or Zein; Rapeseed Protein; Alpha Protein; Pea Tan Protein; legume broad bean protein; cottonseed protein; and sesame seed protein The plant protein according to claim 15, which is a vegetable protein selected from the group consisting of: Pak hydrolyzate.   18. A protein hydrolyzate according to any one of claims 15 to 17 is included. Food products   19. Breast milk substitute ingredients; cheese flavored products; enzymatically produced HVP; protein Enriched diet products; soup-, bouillon- or meat-flavored products; meat extract products; Or a hydrolyzate for improving the production of starter cultures. The food product described in paragraph.   20. A protein hydrolyzate according to any one of claims 15 to 17 is included. It is a non-food product.   twenty one. Pet food, cosmetics or fermentation broth, non-claim 20 Food products.