NL8500956A - Prepn. of improved food flavours - by inactivating yeast and degrading bio-polymers whilst fermenting with microorganisms - Google Patents

Abstract

Food flavour is prepd. by degrading yeast with enzymes. The yeast is inactivated and enzymatic degradation of biopolymers is carried out in conjuction with fermentation with microorganisms. Inactivation is pref. effected by heat treatment. Enzymes used in degradation induce proteolytic, cell wall degrading, glycogen-degrading, RNA degrading or lipolytic enzymes.

Description

R 7012 (R) * *

Applicant: Unilever N.V. in Rotterdam

Title: Food flavoring agents

The invention relates to a method for preparing a food flavoring agent by treating yeast with enzymes.

Such methods are known from the literature.

Yeast autolysates, which are prepared by degrading yeast by the endogenous yeast enzyme material, are well known food additives. Autolysis can be accomplished by incubating the yeast cells at a higher temperature by adding organic solvents, such as ethyl acetate or toluene, by an increased salt concentration or by a combination of these methods. This leads to inactivation of the yeast cells, but the enzymes of the yeast material remain active and are available for the actual degradation process.

The main conversion that takes place during autolysis is the breakdown of proteins into peptides and amino-20 acids. The autolysates thus obtained usually have a typical bitter and yeast-like taste. A further drawback of these autolysates is that they contain 31-ribonucleotides, because the endogenous ribonuclease converts RNA into 3'-ribonucleotides, which do not contribute to the aroma.

It is also known in the art that the enzymatic degradation of yeast is promoted by first breaking down the cell walls with a suitable enzyme, whereby an increased yield of autolysate is obtained (cf. US-A 3 682 778, Kyowa Hakko Kogyo ).

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Furthermore, it is known to increase the yield by using proteolytic enzymes together with the endogenous enzymes (cf. NL-A-7907402, Standard Oil Co).

Finally, NL-A-7601962 (Nestlé) describes the enzymatic degradation of inactivated yeast.

According to the present invention, an improved food flavoring agent is prepared by carrying out the enzymatic degradation of inactivated yeast along with fermentation. The organoleptic properties of the flavors thus obtained are improved, since the bitter taste is absent and there is no, or a less pronounced, yeast-like note present. In addition, organic acids such as lactic acid, succinic acid, etc. also promote taste.

Suitable yeast starting materials belong to the group consisting of the species Saccharomyces, Kluyvero-20 myces, Candida, Torula, Fusarium and Zymomonas. Preferred are the species Saccharomyces, Kluyveromyces, Candida and Torula. More specifically, yeasts such as Saccharomyces cerevisiae, Kuyveromyces lactis, Kluyveromyces fragilis, Kluyveromyces marxianus and Candida utilis can be used.

Before using the yeast enzymes is broken down, it is inactivated, e.g. by heat-treating the yeast for 5-120 minutes at a temperature between 70 and 150 ° C. For inactivation by boiling usually requires an aqueous suspension with a dry matter content of up to 30%. Inactivation is of course also possible by adding chemical products or e.g. by irradiating the yeast.

35

After the yeast has been inactivated, the enzymatic degradation process as well as the fermentation take place in any Λ "Λ r \ '1 * ï-i **' i.

3 R 7012 (R) in sequence or simultaneously, provided that a carbon source is present for fermentation.

The enzymatic degradation of the inactivated yeast 5 is effected by means of suitable enzyme preparations of bacterial, vegetable or animal origin or of yeast. The enzyme preparation used preferably has more than one of the following activities: 1. Proteolytic activity

Preferably, one or more of the following enzymes is used: Pancreatine, Trypsin (from the pancreatic tissue of pigs or cattle), Bromelain (from Pineapple comosus / bracteatus), Ficin, Molsine, Chymotrypsin, Papain (from Carica papaya ), Chymo papain (from Carica papaya), Pepsin 20 (from the gastric mucosa of pigs), Rennine or proteases from Bacillus subtilis, Aspergillus oryzae, Penicillium dupontii, Streptomyces griseus, Mucor miehei / pusillus, pig kidneys, etc.

Depending on the enzyme involved, the incubation takes place at a pH between 2 and 10 and at a temperature between 20 and 80 ° C. Pepsin e.g. has optimal activity at a pH of 2-3, protease from Streptomyces griseus has an optimal activity at a pH of 9-10 and papain is still active at 70-80 ° C.

2. Cell wall degradation activity. Beta glucanase derived from Bacillus subtilis / licheniformis, Penicillium emersonii, Aspergillus niger / oryzae.

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Incubation usually takes place at a temperature between 20 and 70 ° C and a pH between 3 and 7.

3. Amylase or glycogen-depleting activity, e.g.

alpha and beta amylase derived from Bacillus subtilis, Aspergillus spp, which enzymes are commonly used for incubations at a pH of 4-8 and at a temperature of 20-70 ° C.

4. RNA-degrading activity, resulting in 5'-ribonucleotides

Application of e.g. phosphodiesterase, e.g. obtained from malt roots or fungal extracts. These are used at a pH of 3-9 and a temperature of 20-70 ° C.

5. Lipolytic activity 20

Pancreatin or pancreatin lipase that is incubated at a pH of 5-10 and a temperature between 20 and 70 ° C.

25

The enzymatic degradation process of the present invention usually combines several of these enzymatic activities. This can be accomplished by incubating simultaneously with a number of enzymes. A multitude of incubations with different enzymes is also possible, often under different pH and temperature conditions.

Naturally, there are also a number of enzyme preparations commercially available which combine various enzymatic activities, such as pancreatin, etc. When a plurality of enzymatic activities ·; When R 7012 (R) - * 5 is used, incubation may result in the formation of amino acids, peptides, mono- and disaccharides, 5'-ribonucleotides and fatty acids.

Preferably, such a plurality of enzymes are used that the proteolytic activity is accompanied by the degradation of RNA. Cell wall-degrading enzymes and / or lipolytic enzymes are also particularly preferably included.

10

In accordance with the present invention, the enzymatic degradation process is used in conjunction with the fermentation with microorganisms. Preferably, the enzymatic degradation is followed by fermentation, so that saccharides are converted, inter alia, into organic acids, such as lactic acid, succinic acid, etc. It is also advantageous to carry out the enzymatic degradation process simultaneously with the fermentation.

The fermentation with the aid of microorganisms usually takes place at a pH of 4.5 to 7.5 and a temperature of 20-65 ° C, for a period ranging from 4 hours to 14 days. In the practical application of the present invention, use is made of microorganisms commonly used in the preparation of milk products, meat and meat products, fermented vegetables, fermented drinks, bread, piccalilli and sauces, such as: Lactic acid bacteria, e.g. Lactobacillus acidophilus, L. delbrueckii, L. caseï, L. plantarum, L. fermentum, L. brevis, L. buchneri.

Lactic Streptococci, e.g. Streptococcus lactis, Str.

35 cremoris, Str. diacetylactis, Pediococcus pentosaceus, P. cerevisiae, Leuconostoc gracile, L. cremoris.

Yeast, e.g. Saccharomyces rouxii, S. cerevisae, Λ Λ 'Λ Ό Ό h h h R 70 70 70 70 70 70 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 alsmede alsmede combinaties combinaties combinaties combinaties combinaties, as well as combinations of the above microorganisms.

Thus, the yeast extract obtained according to the present invention includes, for example: 20-45 wt% (*) protein material (10-30 wt% peptides and 5-20 wt% free amino acids)? 0.5-6 wt% guanosine 5'-monophosphate? 8-20% by weight of lactic acid.

After the enzymatic degradation process and the fermentation have been carried out, it is recommended to further process the obtained product downstream, such as removing insoluble material (filtration or centrifugation), concentration (evaporation of water, spray drying, oven drying, drum drying or freeze drying , optionally in the presence of a carrier, such as maltodextrin) or pasteurization, e.g. for 5-10 minutes at 80 ° C. Any order is possible.

The food flavoring agent thus obtained can be used as such to impart or enhance the flavor to food, optionally in combination with other flavorings. The combination can be prepared by mixing physically or by chemically reacting to obtain reaction aromas.

The invention also includes the yeast extract prepared by the methods described above.

Thus, one embodiment of the present invention is a method of flavoring foodstuffs by including an aroma in the foodstuff as described above. More specifically (*) calculated on the dry extract 85009c.

7 R 7012 (R), the flavoring is used to improve the aroma of soups, meat products, instant gravy, margarine, baking and frying fat, beverages, bakery products, cheese, confectionery products, etc. The amount of flavor used in the foodstuffs runs varies widely, but usually varies between 0.1 and 10% (calculated as the aroma of dry yeast extract on the foodstuff suitable for consumption). Preferably, these amounts vary between 0.15 and 5%.

10

Example 1 0.67 kg of Saccharomyces cerevisiae (baker's yeast, available from Yeast Brocades, Delft, the Netherlands, with a dry matter content of 30%) was mixed with 0.33 kg of water. The slurry thus obtained was heated to 100 ° C for 10 minutes, then cooled to 60 ° C and the pH adjusted to 4.0 by adding aqueous phosphoric acid.

20 2.2 kg MKC cellulase P 4000 (available from Miles

Kali Chemie GmbH & Co KG, Hannover-Kleefeld, Germany, with an activity of 4000 C / g) was added thereto and the mixture was stirred at 60 ° C for 16 hours. The slurry was now heated to 100 ° C for 10 minutes, cooled to 50 ° C and the pH adjusted to 7.0 by adding a small amount of aqueous sodium hydroxide.

3.0 g of Brew-N-enzyme GPGL (available from Jan Dekker, Wormerveer, Netherlands, with a proteolytic activity of 100 Nü / ml) was then added thereto and the mixture was stirred at 50 ° C for 3 hours. The mixture was then heated to 100 ° C for 5 minutes, 0.5 kg of water was added thereto, and the slurry was cooled to 40 ° C, after which the slurry's pH was adjusted to 5.8 by adding phosphoric acid.

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Now 0.5 wt% of a pre-culture of Lactobacillus buchneri was added thereto. This dense pre-culture of the microorganism on a conventional support had a logarithmic growth phase. The slurry was stirred with this microorganism for 16 hours at 40 ° C.

Then the insoluble material was removed from the slurry by filtration, the filtrate pasteurized at 80 ° C for 20 minutes and concentrated, and a paste with a dry matter content of 60% was obtained. This product was completely water-soluble and had a low sodium chloride content. In organoleptic comparison with yeast autolysate, the product of the present invention was found to have a better, meat-like taste and, moreover, lacked the yeast-like note so common in yeast autolysates.

Example 2 0.2 kg of Kluyvermyces lactis (available from Bel Indus tries, Paris, France) was mixed with 0.7 kg of water in a reaction vessel. The slurry thus obtained was heated to 100 ° C for 20 minutes, cooled to 50 ° C and the pH was adjusted to 4.5 by addition of aqueous phosphoric acid.

2.0 g of MKC hemicellulase (available from Miles Kali Chemie GmbH & Co KG, Hannnover-Kleefeld, Germany, with an activity of 2500 HCU / g) was then added thereto, along with 5 g of malt roots (available from Export Malting "The Netherlands", Wageningen, Netherlands, dry matter content 94%), and the mixture was stirred at 60 ° C for 16 hours. The slurry pH was then adjusted to 5.0 by adding a small amount of sodium hydroxide.

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9 R 7012 (R) 8/8 g of papain (available from Merck & Co, Darmstadt, Germany, with an activity of 30,000 USP-u / mg) and 0.1 g of cysteine were added thereto, then the mixture was left for 3 hours at 60 ° C was stirred. The slurry was then heated to 100 ° C for 10 minutes and 0.33 kg of water was added thereto. The slurry was cooled to 45 ° C and the pH adjusted to 5.8 by adding a small amount of aqueous sodium hydroxide.

Now 0.2% by weight of a pre-culture of Lactobacillus delbrueckii was added to this. This dense pre-culture of the microorganism on a conventional support had a logarithmic growth phase. The slurry was then stirred at 40 ° C for 16 hours.

15

Then the solid was separated by centrifugation and the clear solution thus obtained was pasteurized and concentrated. A yellowish, microbial stable powder was obtained by spray drying on a conventional support.

Example 3 0.2 kg of Torula yeast (available from Attisholz AG, Luterbach, Switzerland, dry matter content 98%) was mixed with 0.8 kg of water in a reaction vessel. The slurry thus obtained was heated to 100 ° C for 40 minutes, cooled to 55 ° C and the pH adjusted to 4.0 by adding aqueous phosphoric acid.

30 2.0 g MKC Cellulase P 4000 (available from Miles Kali Chemie GmbH & Co KG, Hannover-Kleefeld, Germany, with an activity of 4000 Cü / g) and 1.0 g bromelain (available from Sigma Chemie GmbH, Taufkirchen, Germany, with an activity of 2000 u / g) were added thereto, after which the mixture was stirred at 55 ° C for 8 hours. The slurry was then cooled to 40 ° C, Λ 'Λ Λ * 1¾ ü:} ‘,. J t! R 7012 (R) was added to this 0.25 kg of water and the pH was adjusted to 5.8.

0.8 wt. Of a pre-culture of Lactobacillus 5 plantarum was now added. This dense pre-culture of the microorganism on a conventional support had a logarithmic growth phase. The slurry was stirred at 40 ° C for 16 hours and then cooled to 25 ° C.

Then 0.1 wt% of a fresh, dense pre-culture of Saccharomyces rouxii was added thereto at 25 ° C and the slurry was stirred at this temperature for 48 hours. The slurry thus obtained was then processed as described in Example 2 to obtain a yellowish powder.

Example 4 0.2 kg Saccharomyces cerevisiae (available from Nedalco N.V., Bergen op Zoom, The Netherlands, dry matter content 97%) was mixed with 0.8 kg water in a reaction vessel. The slurry thus obtained was heated to 100 ° C for 10 minutes, then cooled to 50 ° C and the pH was adjusted to 8.0 by addition of aqueous sodium 25 hydroxide.

1.0 g of pancreatin (available from Merck, Darmstadt, Germany, with an activity of 30,000 FIP-U / g [lipase]) was added to the slurry and this mixture was stirred at 60 ° C for 2 hours. the slurry was then adjusted to 4.0 by adding a small amount of aqueous phosphoric acid.

10 g of Brew-N Enzyme Filtranase L 5 (available from Jan 35 Dekker, Wormerveer, The Netherlands, with an activity of 9500 Endo-beta-1,4-glucanase µ / ml) was combined with 0.8 g of ficin (available from Sigma Chemie GmbH, Taufkirchen, III / 3 ^ R 7012 (R) 11

Germany, activity 1.5 u / mg protein) was added thereto and stirred at 60 ° C for 6 hours. The slurry was then heated to 100 ° C for 10 minutes, cooled to 40 ° C and the pH adjusted to 5.8 by adding a small amount of sodium hydroxide.

Now 0.5% by weight of a pre-culture of Streptococcus diacetylactis was added thereto. This dense pre-culture of the microorganism on a conventional support had a logarithmic growth phase. 3.0 g of Brew-N-zym / GPGL (available from Jan Dekker, Wormerveer, The Netherlands) was added to this fermentation. The slurry was then stirred at 40 ° C for 16 hours.

15

The slurry was then processed as described in Example 2 to obtain a fine, yellowish powder.

20 Example 5

A meat flavor was prepared by mixing the following ingredients: 200 g yeast extract paste obtained according to Example 1 10 g glucose 2 g cysteine 3 g thiamine HCl salt 2 g tallow 30

The mixture was refluxed for 90 minutes. After the mixture was cooled to 40 ° C, 200 g of maltodextrin and 200 g of water were added thereto.

The solution thus obtained was spray dried.

After the spray dried powder in an amount of 1.5% to a 0.5% aqueous solution of table salt

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35 12 R 7012 (R), t 6 was added, which gave it an excellent taste of boiled beef, which closely resembled the taste of real beef broth.

Example 6 2.5 g of a spray dried yeast extract as prepared according to Example 4 was added to the following dry soup product: 10 10 g sodium chloride 2.5 g monosodium glutamate 6 g tallow 20 g vermicelli 4 g dried chopped onion 1.5 g dried, chopped carrots 0.25 g of a herb and spice mixture

The whole was taken up in 1 liter of water and boiled for 20 minutes. A group of subjects found the soup to be rounded, spicy and sweet beef-like.

Example 7 2.0 g of a spray dried yeast extract as described in Example 3 was added to the following fish sauce: 8 g skimmed milk powder 30 10 g wheat flour 1 g sodium chloride 1 g monosodium glutamate 80 g water 35 After the mixture was up to 100 ° C heated, a sauce with a full aromatic fresh taste was obtained, which was successfully used in the ratio of 2 parts sauce to 1 part boiled cod.

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Example 8 0/1 part of spray dried yeast extract as prepared according to Example 2 was added to the water phase of a margarine / which water phase consisted of: 5% water 10% skim milk 1% sodium chloride.

10

This mixture was adjusted to pH 5.6. The fat phase consisted of 10% solid phase of the margarine (hardstock), obtained by transesterification of hydrogenated soybean oil (melting point 43 ° C), 42% palm oil (melting point 58 ° C) and 90% liquid sunflower oil in the usual manner processed in a margarine. When used as a table margarine, a cool, creamy impression was obtained.

When used as a margarine for baking and roasting, a sweet flavor reminiscent of cream-20 butter developed at 140 * 0.

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Claims (15)

Process for preparing a food flavoring agent by breaking down yeast with enzymes, characterized in that yeast is inactivated and the enzymatic degradation process of biopolymers is carried out in addition to fermentation with micro-organisms. Method according to Claim 1, characterized in that further work-up takes place afterwards. Method according to Claim 1 or 2, characterized in that the yeast starting material belongs to the group consisting of Saccharomyces, Kluyveromyces, Candida and Torula. Method according to Claim 1, 2 or 3, characterized in that the inactivation takes place by application of heat. 5. Process according to one or more of the preceding claims, characterized in that proteolytic enzymes are used in the enzymatic degradation process. 6. Method according to one or more of the preceding Claims, characterized in that cell wall-degrading enzymes are used in the enzymatic degradation process. 7. Process according to one or more of the preceding Claims, characterized in that glycogen-degrading enzymes are used in the enzymatic degradation process. 8500958 R 7012 (R) "" "* Method according to one or more of the preceding Claims / characterized in that such RNA-degrading enzymes are used in the degradation process that 5'-ribonucleotides are formed. 5 Method according to one or more of the preceding Claims, characterized in that lipolytic enzymes are used in the enzymatic degradation process, Process according to one or more of the preceding Claims, characterized in that after the enzymatic degradation process fermentation by means of micro-organisms. Process according to one or more of Claims 1 to 9, characterized in that the enzymatic degradation process and the fermentation by means of microorganisms are run simultaneously. Method according to one or more of the preceding Claims, characterized in that a microorganism is used in fermentation to form lactic acid. Process according to one or more of Claims 1 to 11, characterized in that a yeast is used in the fermentation. 14. Yeast extract obtained by a method as described in one or more of the preceding Claims. 15. A method of preparing food flavoring agents, characterized in that a yeast extract according to claim 14 is used. - - '^, »z Λ' * ·. · Λ *« V - .. «> V