DE1692709C - Process for the production of condiments from the microbial masses of fermentation broth - Google Patents

Description

After adjusting the mixture to pH 5.5 to 6.5, concentrated in vacuo at 45 to 60 ° C will. While the known process for the production of seasonings or Fleiixexttrakte from Microbial masses in bacteria as microbial mass have so far not led to the desired success, since as a result of the greater resilience of ZeIlwäp.de of the bacteria, the enzymatic hydrolysis resulted in only a low yield the pretreatment according to the invention has the desired success. At the same time leads to the invention Worship to an increased yield. Those produced by the process of the invention We have large amounts of essential amino acids and various other nutrients stoOYn. z. B. vitamins, nucleotides, nucleosides, per NitVwert.

The invention is based on the following Bciriclen explained in more detail:

example 1

00 1 fermentation broth, which was prepared by cultivating Microco -us glutamicus ATCC 13761 in the usual manner, was concentrated to a pH of 4.5 and centrifuged, whereby 5.5 kg of microbe mass (water content 75.5 ⁿ / o) received. 35 l of a 0.05 μm aqueous sodium carbonate solution were added, with cell milk being formed. This was heated to 55 ° C. for 2 hours with stirring in order to destroy the cell winds and membranes. The pH of the liquid was then adjusted to 7.2 by adding 2η aqueous hydrochloric acid and the solution was heated to 105 ° C. for 10 minutes. After cooling to 55 ° C., 7.5 g of protease, ie in an amount of 1% of the total protein content, were added to the solution; then the liquid was hydrolyzed at 50 to 55 ° C for 16 hours. The hydrolyzed liquid contained 2.7 g l total nitrogen and 2.2 g / l glutamic acid. In this fluid, 82% of the cellular protein was broken down with the formation of amino acids and peptins. If the decomposed liquid had a bitter taste, the following procedure was used to remove this bitter texture and the coloring substances: the liquid was adjusted to a pH of 4.8 by adding 2% aqueous hydrochloric acid and filtered to obtain it an undecomposed part and a filtrate. The undecomposed part was washed with 21% water. The filtrate and the washing water obtained were combined to give 36.5 liters of a clear liquid. This was sent at a flow rate of 2.0 through a resin column which was charged with 1.81 of a decolorizing resin in the Cl form in a volume of ¹ Ao to the volume of the liquid. The elution was over after about 10 minutes. The resin was then washed with 4 liters of water. Eluate and wash water gave 401 combined liquid with no bitter taste. After adjusting the pH by adding 2N sodium hydroxide solution, the liquid was concentrated at 50 to 60 ° C under reduced pressure, and 780 g of a seasoning paste were obtained, which had greatly improved properties and the following composition:

Analytical results of the seasoning paste

Water (° / o) 12.5

NaCl (Vo) 15.2

Amino acids (%>)

Glutamic acid 8.0

Aspartic acid 1.8

Threonine 0.8

Serine 0.8

Proline 0.4

Glycine 0, f>

Alanine 3.5

Valine 1.0

Methionine 0.5

Isoleucine 0.8

Tyrosine 0.4

Leucine 0.9

Phenylalanine 0.8

Lyline 1.1

Arginine 0.9

5'-inosinic acid (%) 2.0

Vitamins (y / g)

Riboflavin 20.5

Thiamine 5.6

Nicotinic acid 48.8

Pantothenic acid 28.0

Biotin 0.01

Pyridoxine 2.0

Inositol 410.0

Folinic acid 1.8

pH of the aqueous liquid 5.5

Example 2

1001 of a broth formed by fermenting 5'-inosinic acid by culturing Brevibacterium ammoniagcnes KY 3454 in the usual way, was adjusted to a pH of 4.5 and, after centrifugation, gave 8.4 kg of cellular mass ( Water content 70.2%>); 52 liters of a 0.03 η aqueous sodium hydroxide solution were added to this and a cell milk was obtained. This was heated to 60 ° C. for 1 hour with stirring in order to loosen the cell walls and membranes; then added to 2 η aqueous hydrochloric acid is added, to adjust the pH to 7.0, and then heated 10 minutes 105 ⁰ C. After cooling to 50 ° C were added to the liquid 12.5 g (ie, I ° / o based on the protein substrate) Streptomyces protease added to decompose the cell protein at 50 to 53 ° C. Eight hours after the start of decomposition, an additional 12.5 g of the same protease was added to the solution and hydrolysis proceeded for a further 8 hours (total time 16 hours). This enzymatic treatment converted 80 to 82% of the cell protein into amino acids and lower peptides. After the completion of the decomposition, the liquid was heated at 105 ° C for 5 minutes to stop the reaction and then allowed to cool. The solution was adjusted to a pH of 4.6 by adding aqueous 2η hydrochloric acid and nitrated. 50 l of the filtrate were obtained, separated off from the unreacted material. The latter was washed with 31% of water. Filtrai and wash water were combined and sent through a resin column which

■ äi
Iu

: ii in
Ir
ir

λ ¹ ! ie
ir
O

was charged with 2.5 l of a resin in its Cl form, with a flow rate of 2. Nach The elution was over for hours. The column was then washed with 5 liters of water. One received 1 of a combined eluate. This was by adding 5 aqueous 2 N Natron'.auge to one pH adjusted to 5.5 and concentrated under reduced pressure and gave 1.4 kg of a seasoning paste, which contained glutamic acid, sodium 5'-inosinate, and various other amino acids; they ok had excellent taste-improving properties. The following table shows the analysis values this seasoning.

Analytical results of the seasoning paste ij

Water ( ⁰ Zo) 12.5

NaCl ( ⁰ Zo) 15.5

Total nitrogen ( ⁰ Zo) 10.5

Amino Acids ( ⁰ Zo)

Alanine 3.0

Lysine 0.8

Leucine 0.8

Glutamic acid 1?., 5 25

Aspartic acid .... _; 1.3

Valine 0.8

other amino acids were also detected

Vitamins (γ / g)

Thiamine 2.1

Pantothenic acid 2.5

Pyridoxine 0.8

Folinic acid 0.9

Riboflavin 15.5

Nicotinic acid 25.6

Inositol 350.0

pH of the aqueous liquid 5.5

Example 3 ■

100 1 of a fermentation broth (content per 100 ml: 1.2 g Gl ".-: - amic acid, 0.58 g alanine, 0.5 g aspartic acid, 0.5 g glutamine, 0.1 g lysine, 0.15 g valine, 0, 13 g of leucine, 1.4 g of dry cells, also proteins, peptides, vitamins, sugar and the like), which had been obtained by separating the substantial amounts of glutamic acid from the fermentation solutions of the glutamic acid fermentation, were mixed with sodium carbonate to bring about a concentration of 0.03 η, and concentrated in vacuo at 50 ° C. in order to remove impurities such as ammonia, for example 50 l of the concentrated liquid, which was heated to 55 ° C. for 2 hours, by adding hydrochloric acid to one The pH was adjusted to 7.2 and heated for 10 minutes to 105 ° C. After cooling, the liquid was transferred by adding the coloring matter. The removal of Jer _{R (} ^

of poorer taste a ^ _Ηο

^m W h ZtZ of HCl to a pH of

was adjusted by ^additives * ^U ™ _{"k" Tre} retr of the filtrate of 4.8 and for the purpose of Ire g ^ ^

Wash the undecomposed R ^UC ,, fA """". One received

s.gke g ^

; and one united washing

e ^ chne * schnappsve
and had the following composition ".

Together

water

ication of the wort (in weight)

What, _ _lü

Total embroidery stolt 10

Alin

Alanine

12.5 ^e / o _lü5 " _{/ o}

3 «/» 20 ° / »

Leucine

Thiamine

Pantothenic acid

Pyridoxine

Folinic acid

different amino acids were also included

other

NaC 1

Glutamic acid

Aspartic acid

Valine Γ
Riboflavin
Nicotinic acid
Inositol

4.3 yig

37.1 yZg 1.5 yig 1.8 y / g

12.8 ⁰ zo

30.1 «/ 0

10.1 "Zo

4.0 «/»

16.4 yl%

66.6 yig

375.0 yZg

Example

l 4

from

Gc

16

a broth obtained by fermentation of amic acid and which is contained in 100 ml of 1.2 kg of cTmaminic acid, 0.5 g of alanine, 05 g of aspartic acid, 0.3 g of glutamine, 0.1 g of lysine. 0.15 g valine 0.13 g leucine 1.4 g microbes (measured in the dried state) as well as peptides, proteins, vitamins, sugars and the like were mixed with sodium carbonate in order to set a concentration of 0.03 η 50 C concentrated, and 50 1 of the concentrated Fluss.gke.t were obtained. Hydrochloric acid was added to this liquid to make the. _{ulf 7} -> set, and then heated to 105 ° C for minutes. After

G
pH wide

LlIl / MmillMin wuiui. >Ji> - · ■ «■■ (- · ■.. ..« 4; ,,, .. ,,., Allt HJ '^ LIIIlIiI- I1i "-H UCIII

on 9 g of protease corresponding to 1 percent by weight of the 6c was si. Minut π au. ^ ^ ^

k-velvet-P, otein substrate enzymatically decomposed and cooling, f gU mar, ^ ^d ? J _R ⁸ _ührcn ^b _{in lfiStun} .

, Heated to 50 to 55 ° C. for 6 hours with stirring. tease hmzu u ^ J ^} ^L ™ " _{H droiysat was} with

As a result of the decomposition, cell proteins and free den, at S (J ^ b- «■ · 'and

^^ iciSrSS S ^ T '^^ tSS a ₅ SSSVai cS'5. FiHr .. and 2.5 1 undecomposed

and various other S contained in the cell bodies became {• plump cluicit. The proteins were in Amino acids and low peptides to 82 to

85 ⁰ Zo

IlllllV.it. I'll · .. ~. . -

Residue (water content 70.2 ⁰ Zo). To the residue unz.ersetzten (35 ⁰ Zn) were added 1.3 kg of concentrated hydrochloric acid, '' ^J -

and then became the

Residue hydrolyzed by 16stündigcs heating to 105 ^'1 C to obtain a hydrolyzate was obtained in which the ratio of Gesamtstiekslolf to Aminostickstolf was 2.1.

2 liters were added to the hydrolyzed liquid Water and filtered this, a residue being obtained in addition to the filtrate. The latter was with 2 1 water washed. Wash water and filtrate gave 7 liters of liquid, which under reduced Pressure were concentrated to 1.5 liters. The concentrated solution was mixed with the liquid that came with the enzymatic treatment was obtained, combined and adjusted to a pH of 6.4 with sodium hydroxide solution. This liquid was then concentrated under reduced pressure at 45 to 50 ° C and received a seasoning in paste form. The product obtained had excellent skin-improving properties Properties and contained various amino acids, peptides, organic acids. Vitamins and the like and was suitable for all seasoning purposes.

Example 5

100 l of a fermentation broth of 5'-inosinic acid, obtained by culturing Brevibacterium ammoniagenes KY 3454 in the usual way, was adjusted to a pH of 4.5 and, after centrifugation, gave 8.4 kg of mass (water content 70.2 ° /O). In order to obtain cell milk, 52 liters of 0.03 η sodium hydroxide solution were added to this. This express milk was heated to 60 "C for 1 hour with stirring in order to dissolve the cell walls and membranes, then aqueous 2N hydrochloric acid was added to adjust the pH to 7.0, and then to 105 ^{3 for 10 minutes} C. After cooling to 45 "C, 7.7 g of Strcptomyces protease were added to this liquid in order to decompose the cell protcin at 45 to 47" C in 10 hours.

At the end of the decomposition, aqueous 2N hydrochloric acid became added to adjust pH to 4.5 and filtered. 50 l of filtrate and 4 kg were obtained undecomposed residue (water content 70.2%). The HiItrat was sent through a resin column that included 1.7 l of a decolorizing resin in its Cl form was charged at a flow rate from 2 through. The elution was over after 15 hours. The column was then filled with 3 liters of water washed and this wash water added to the eluate. This resulted in 531 liquid.

The undecomposed residue in amounts of 4 kg was treated with 1.65 kg of concentrated hydrochloric acid (35%) added and hydrolyzed at 100 C for 13 hours. The resulting hydrolyzed liquid contained besides Amino acids large amounts of peptides. The ratio of total N-3 n-amino-N was 2.6. The 21 water were added to the hydrolyzed liquid, cooled, and a filtrate and a residue were obtained after filtration. The latter was with 21 water were added and after filtration gave 8 l of filtrate, which was concentrated to 21. The concentrated filtrate was mixed with aqueous 2η sodium hydroxide solution in order to adjust the pH to 4.0; the filtrate was then made up with water until an amount of liquid of 5 liters was obtained and treated it with 60 g of activated charcoal to remove the coloring substances and bitter substances. The liquid was filtered and 4.5 liters of filtrate were obtained. which with the aforementioned eluate, the obtained by treatment with the decolorizing resin. The combined liquid was adjusted to a pH of 6.2 with sodium hydroxide solution and concentrated in vacuo at 45 to 50.degree. and 2.1 kg of wort in paste form were obtained. The wort obtained contained sodium 5'-inosinic acid and Amino acids such as glutamic acid, peptides, vitamins and the like, and had a meat-like taste. Different Tests including a functional test showed that the condiments were excellent in improving taste Hold properties.

The following table shows the analytical results of the wort of this example:

Analysis of the wort

Water 13.7 ⁿ / n

NaCl 10.6 / «

Thiamine 10.0 - / g

ϊο pantothenic acid 40.0; / g

Pyridoxine 3.0 v / g

Folinic acid. ·. 3.0; - / g

Total nitrogen 7.7%

5'-inosinic acid 6.1 ^fl / o

»5 riboflavin 31.0 _; /G

Nicotinic acid 0.2 y / g

Biotin 0.03 -; u

Inositol 540.0- / g '

This wort alone can, if desired, be processed with the HCl hydrolyzate of the insoluble function which can be produced by the process according to the invention.

Example 6

100 l of a fermentation broth obtained by cultivating Micrococcus glutamicus ATCC. 13761 was obtained in the usual manner, was adjusted to a pH value of 4.5 and gave 5.5 kg after centrifugation. Microbial mass (water content 73.5 ⁰ Zn) 35 1 aqueous 0.05 N sodium carbonate solution were added for the purpose of resounding cell milk. This cell milk was heated to 55 (for 2 hours while stirring. Then the pH of the liquid was adjusted to 7.2 with aqueous 2N acid / acid, and it was heated to 105 ° C. for 10 minutes after cooling 55 ° C, the liquid wedge was treated with 7.5 g of a protease, ie with 1% based on the total protein content, and hydrolyzed for 16 hours at 50 to 55 ° C. The hydrolyzate contained 2.7 g 1 total nitrogen and 2.2 g / l Glutamic acid: In this liquid, 82% of the cellular protein was broken down with the formation of amino acids and peptides.

Because the hydrolyzate had a bitter taste. was used to remove the bitter taste and the coloring substances proceeded as follows: The liquid was by the addition of aqueous 2N hydrochloric acid was adjusted to pH 4.8 and to obtain a filtrate and an undecomposed one Partly nitrided. The last was washed with 2 liters of water. The filtrate and wash water gave 36.5 liters of a combined clarify liquid, which is then passed through a resin column containing 1.81 of a decolorizing resin was packed in its Cl form, in Vjn of volume of the liquid to be decomposed, with a flow velocity from 2.0 was sent. The elution was complete after about 10 hours. One washed

3D? 623/270

I 692 709

the resin with 4 l of water, combined eluate and wash water and obtained 50 l of liquid which was free from a bad taste. After the pH value had been adjusted to 5.5 with aqueous n-sodium hydroxide solution , the solution was concentrated under reduced pressure at 50 to 60 ° C., and 780 g of a pasty substance were obtained which had excellent taste-improving properties and the following composition:

Analytical results of the seasoning paste

Water 12.3%>

Glutamic acid 11.8%>

Thiamine 7.2 γ / g

Pantothenic acid 62.0 γ / g

Pyridoxine 3.2 γ / g Folinic acid 3.1 γ / g

NaCl 11.5%,

Total nitrogen 12.3%> Riboflavin 27.3; </ g Nicotinic acid 112.0 γ / g

Biotin 0.01 γ / g

Inositol , 625.0 γ / g

Example 7

The undecomposed part, which was obtained in the same manner as described in Example 1 in an amount of 5.0 kg (water content 72.5%), was admixed with 2.0 kg of concentrated hydrochloric acid (35%). The concentration of hydrochloric acid in the mixture was about 3.5%. It was hydrolyzed at 111 ° C for 14 hours and a decomposed liquid was obtained with about 2.0 APL-Wcrt, ratio of total nitrogen sticks / u aminostickslolT, which proved that the liquid was lower in large quantities Contained peptides and amino acids. After cooling, this liquid wedge was filtered in order to separate the sediment from the filtrate. 4 l of water were added to the filter cake, and the washing water was combined with the aforementioned filtrate to form 8.5 l of liquid, which was made up to 10 l with 1.5 l of water. They were then concentrated to 5 l in vacuo. To adjust the pH to 4.5, sodium carbonate and sodium hydroxide were added to the concentrate, and the concentrate was made up to 10% of liquid with water. 100 g of activated charcoal were then added to the liquid and the mixture was stirred for 1 hour in order to remove impurities such as coloring substances and the taste of the beer. After filtration, 9.5 l of fillrate were obtained. 101 of the resulting filtrate were combined with 5 liters of the decomposed liquid in a manner similar to that described in Example 3. The combined liquids were treated in the manner described in Example 1 to give 2.2 kg of a combined seasoning in paste form, which was found to be an excellent flavor-improving agent by various tests including Gesehmack index.

Claims (2)

1 2 In addition, the production of meat extracts from aqueous yeast is known (cf. British patent specification: patent claims: 861 612) 1. Process for the production of condiments from a by the Miroben masses of the fermentation broth, which at 5 with the from ^ ... Extraction of amino acids or nucleic acid obtained hydrolyzate combined and getrocKnei corresponding compounds by fermentation contains before the enzymatic hydrolysis by heating in the existing microbe mass, an alkali io an alkaline salt solution and salt of a weak acid and / or an. Alkali to clean the hydrolyzate by means of ion exchangers or the like hydroxide to adjust a concentration this I is supplied from 0.02 to 0.1 η, the liquid is thus the microbe mass 1 to 3 hours with stirring to 50 to 80 ° C. Yeast on the one hand and bacteria heated, the pH value then to 6, 0 to 8.0 15, however, have fundamentally different cell body is set, the liquid 3 to 10 minutes in particular, have the Walls of these cells heated to 80 to 130 C, after cooling with Pro- different compositions. In yeast icasc added and to break down the protein 6 to the composition consists of mannan and is kept at 50 to 60 ° C for 48 hours, after glucan, while the firmer cell walls of Bak cool the pH to 4.5 to 4, As is well known, 8 ao reries are made up of peptides and murine, the liquid is filtered and the acidic acid (murein) is made up. The application of the stock is washed out with water, from the known measures for the production of condiments combined washing water and filtrate with the help of a or meat extract from microbial mass, the bitter substances bacteria as microbial mass have not yet been removed in resin columns or with activated charcoal, the eluate or 25 desired success, because due to the larger cons-filtrate, the cell walls of the bacteria are able to stand up to a pH value of 5.5 to 6.5 and the cell walls of the bacteria are corrected at 45 to 60 "C in a vacuum. enzymatic hydrolysis is only a low yield. 2. The method according to claim 1, characterized in that it is basically known (see franking marks, that the French patent 1 369 620), fermentation broths from the Remaining residue several hours with con-amino acid production, which is still the microbial center Hydrochloric acid (35%) contains up to a mass to be mixed with calcium hydroxide ratio of total nitrogen to amino nitrogen and heat the mixture, thereby removing the bacteria from about 2 is hydrolyzed, the hydrolyzate cells are to be destroyed. However, this has to diluted with water, filtered, the residue with 35 Continuation of a method of the initially described-water is washed, washing water and filtrate have not contributed to the genus so far, be concentrated, this concentrate with the The invention is based on the object obtained in the enzymatic treatment a method of the type described above Solution combined and this mixture after working with a microbial mass of bacteria, position to a pH value of 5.5 to 6.5 in 40. To solve this problem, the invention teaches Vacuum is concentrated at 45 to 60 "C. That of the aqueous fluid that is made up of bacteria contains existing microbial mass, an alkali salt of a weak acid and / or an alkali hydroxide for setting a concentration from 0.02 to 45 0.1 η is supplied, the liquid 1 to 3 hours heated with stirring to 50 to 80 ⁿ C, the pH value then adjusted to 6.0 to 8.0, the liquid 3 to 10 minutes to 80 to 130 ⁰ C, after cooling mixed with protease and to break down the The invention relates to a method for keeping 50 proteins at 50 to 60 ° C for 6 to 48 hours Production of condiments from the microbial masses is done after cooling the pH to 4.5 up of the fermentation broths, which ceased during the recovery of amino-4,8, the liquid filtered and the residual acids or nucleic acids corresponding compound is washed out with water, from the digestion arise through fermentation. In a known process of the resin column described, or with activated charcoal, the bitter substances combine with the washing water and filtrate The genus (cf. Belgian patent specification 659 731) is that and dyes are removed, the eluate or Microbial mass a yeast, namely a yeast filtrate is adjusted to a pH of 5.5 to 6.5 exclusion after thermal inactivation and is concentrated at 55 to 60 ° C in a vacuum. Yeast enzymes under suitable pH temperature conditions. A preferred embodiment of the invention is Kind treated with a proteolytic enzyme Bo characterized in that the after filtering and the hydrolyzate to inactivate the added remaining residue for several hours with kon-En / .yms Again it can be used from concentrated hydrochloric acid (35%) up to a ratio a yeast from a fermentation broth from total nitrogen to amino nitrogen of about 2 to be gone. The enzymatic hydrolysis is hydrolyzed, the hydrolyzate is mixed with water a temperature between 50 and 60 "C at 65, filtered, the residue washed with water a pH between 6 and 8 during 6 to, wash water and filtrate are concentrated, Hours running. After separating the solid this concentrate with that of the enzymatic substances, the filtrate is thickened in a vacuum. Treatment obtained solution combined and this