DE1692709B - Process for the production of condiments from the microbial masses of fermentation broth - Google Patents

Description

45

The invention relates to a method for producing condiments from the microbial masses the fermentation broth, the compounds corresponding to the production of amino acids or nucleic acids arise through fermentation.

In a known method of the type described (see. Belgian Patent 659 731) is the Microbial mass a yeast, namely a yeast elimination after thermal inactivation of the Treated yeast enzymes with a proteolytic enzyme under suitable pH temperature conditions and the hydrolyzate is heated again to inactivate the added enzyme. You can also use a yeast originating from a fermentation broth can be assumed. The enzymatic hydrolysis will at a temperature between 50 and 60 ° C at a pH between 6 and 3 for 6 to Hours running. After the solids have been separated off, the filtrate is concentrated in vacuo.

In addition, the production of meat extracts is known (see British patent specification 861 612) aqueous protein extracts of yeast. Here an extract obtained by proteolytic enzymes is used combined with the hydrolyzate obtained from the residue by acid digestion and dried. The measure (cf. British patent specification 933 828), yeast, also belongs in this context to digest before enzymatic hydrolysis by heating in an alkaline salt solution and To purify the hydrolyzate by means of ion exchangers or the like. - As part of all these known measures So the microbial mass is ultimately a yeast. Yeast on the one hand and bacteria on the other however, have vastly different cell bodies. In particular, the walls of these conductors show different compositions. In the case of yeast, the composition consists of mannan and Glucan, while the firmer cell walls of bacteria are known to consist of peptides and muramic acid (Murein) exist. The application of the known measures for the production of condiments or meat extracts from microbial mass did not lead to the desired microbial mass in bacteria Success, because of the greater resistance of the cell walls of the bacteria enzymatic hydrolysis resulted in only a low yield.

However, it is basically known (see. French Patent 1,369,620), fermentation broths from the To add calcium hydroxide to amino acid production, which still contain the microbial mass and heating the mixture, thereby destroying the bacterial cells. However, this has to Continuation of a method of the type described above has not yet contributed.

The invention is based on the object of a method of the type described at the outset to work with a mass of microbes made up of bacteria. To solve this problem, the invention teaches that the aqueous liquid containing the bacterial mass of microbes is an alkali salt a weak acid and / or an alkali hydroxide to adjust a concentration of 0.02 to 0.1 η is added, the liquid is heated to 50 to 80 ° C for 1 to 3 hours while stirring, c-, r pH value then adjusted to 6.0 to 8.0, the liquid 3 to 10 minutes at 80 to 130 ° C, after cooling treated with protease and kept at 50 to 60 ° C for 6 to 48 hours to break down the protein the pH is adjusted to 4.5 to 4.8 after cooling, the liquid is filtered and the residue is washed out with water, from the combined wash water and filtrate using a Resin column or with activated carbon the bitter substances and colorings are removed, the eluate or The filtrate is adjusted to a pH of 5.5 to 6.5 and concentrated at 55 to 60 ° C in vacuo. A preferred embodiment of the invention is characterized in that after filtering remaining residue several hours with concentrated hydrochloric acid (35%) up to a ratio hydrolyzed from total nitrogen to amino nitrogen by about 2, dilute the hydrolyzate with water filtered, the residue is washed with water, the wash water and the filtrate are concentrated, this concentrate is combined with the solution obtained in the enzymatic treatment and this

rpmisch after adjusting to a pH value of 5.5 wfi5 in vacuo at 45 to 60 ° C concentrated ■ -Λ 'during the known processes for manufacturing! SLIC of seasoning or Fleischcxtrakten from Srobenmassen in bacteria as a microbial mass Ser not produce the desired success led there - fnioe the greater resilience of Zeil- "sands of bacterial enzymatic hydrolysis wines low yield result, had reached T _e: pretreatment according to the invention the desired ti Frfol- same Proceeding to the invention results in increased yield the invention according to In. .. A method Szen produced have larger quantities of essential amino acids, and various other nourishes ffen ζ Β vitamins, nucleotides Nucleosides Pepf.den and the like, and are therefore seasoning having a very high ^Ν ΪΓfolgenden the invention with reference to detailed examples will be explained: ^2O -

example 1

1001 fermentation broth, which was prepared by cultivating Micrococtus glutamicus ATCC 13761 in the usual manner, was adjusted to a pH of 4 pounds and centrifuged, giving 5.5 kg of microbe mass (water content 75.5% by volume ) . One added «Tdner 0 05 η wäßripen N _? <rium carbonate solution II whereby a cell layer formed. This was? Hours of stirring to damage the cell walls and membranes heated to 55 ° C. Then riinderFliquid ^ chZusa.v ^^ wa; «- Analytical results of the seasoning paste

Water (° / o)
NaQ (Ve)

Amino acids (° / o)

12 5

8.0 i.8

Glutamic acid

Aspartic acid

Threonine

Serine

Proline

Glycine

Alanine,

VaUn

Methionine «, -

Isoleucine 0.8

Tyrosine 0.4

Leucine 0.9

Phenylalanine 0.8

1.1

_o ' _{) 4 06}

Q5

Lyline

Arginine

5'-inosinic acid (° / o)
Vitamins (y / g)

Riboflavin

Thiamine

Nicotinic acid.
Pantothenic acid

Biotin

Pyridoxine

Inositol

Folinic acid

0.9 2.0

20.5 5.6

48.8

28.0 0.01 2.0

410.0 1.8

pH of the aqueous

Example 2

Liquid ->

1 of a broth formed by fermenting 5'-ino, namely by growing ■ '~ - * ~ - ^: " ^m ommnniagenes KY 3454 in

sic acid

put the liquid in a diueren,

the following procedure was used to eliminate this bitter taste and the coloring substances: The liquid was adjusted to a pH of 4.8 by adding 2 η aqueous hydrochloric acid filtered to obtain an undecomposed part and a filtrate. The undecomposed part was with Water washed. The filtrate and the resulting wash water were combined to give 36.5 a clear liquid received. These were sent

'- ^: - "i;« Voit vnn 2.0 through 5:

. __ .iimx.v, _. . IC UUU -u.w .. ^

to solve; 2 η aqueous hydrochloric acid was then added in order to adjust the pH to 7.0, and the mixture was then heated to 105 ° C. for 10 minutes. After cooling to 50 ° C., 12.5 g (ie 1 ° C.) were added to the liquid / o based on the protein substrate) Streptomyces protease added to decompose the cell protein at 50 to 53 ° C. 8 hours after the start of the decomposition, an additional 12.5 g of the - "- ⁿ - *" ocp were added to the solution, and the -, α "" itprPT 8 hours from (Ge

ta

wa,

After about 10 minutes it was

sannzen iu oiunu »,.,.

80 to 82 ⁰ As of the cell protein were converted into amino acids and lower peptides. After the end of the decomposition, the liquid became --- ~~ -,.: ._ * " _{m the} Rektion

position eic * [m - ». ^. ^ o„ _ Addition of aqueous

caustic was the liquid at 50 to 60 ° C below to a pH value \ "...,
reduced pressure concentrated to give 65 to yield 50 1 filtrate, separated sale ... _.
780 g of a seasoning paste, which is greatly improved material. The latter was treated with 31 water and the subsequent washing together. The filtrate and wash water were combined, settled: and they were sent through a resin column which

1

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 10 hours. The column was then washed with 5 liters of water. One received 551 of a pooled eluate. This was by adding aqueous 2 N sodium hydroxide solution to a 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 ίο had excellent taste-improving properties. The following table shows the analysis values this seasoning.

Analytical results of the seasoning paste

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 12.5

Aspartic acid 1.3

Valine 0.8

other amino acids were also detected

Vitamins (yZg)

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 l of a fermentation broth (content per 100 ml: 1.2 g glutamic acid, 0.58 g alanine, 0.5 g aspartic acid, 0.5 g glutamine, 01 g lysine, 0.15 g valine, 0.13 g leucine, 1.4 g dry cells, also peptides, 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 achieve a concentration of 0.03 η and concentrated in vacuo at 50 ° C to remove impurities such. B. ammonia to remove. Mari received 50 l of the concentrated liquid, which was heated to 55 ° C. for 2 hours, adjusted to a pH of 7.2 by adding hydrochloric acid and heated to 105 ° C. for 10 minutes. After cooling, the liquid was enzymatically decomposed by adding 9 g of protease corresponding to 1 percent by weight of the total protein substrate and heated to 50 to 55 ° C. for 16 hours with stirring. As a result of the decomposition, cell proteins and free proteins were hydrolyzed to amino acids and peptides, and various other substances such as e.g. B. Vitamins contained in the cell bodies were thoroughly eluted. The proteins were broken down into amino acids and lower peptides to 82 to 85 ⁰ Zo 709 ¹ I

¹ 6

convicted. The coloring matter and the bitter taste were removed from the liquid as follows: The liquid was adjusted to a pH of 4.8 by adding HCl and filtered to separate the filtrate from the undecomposed residues. The residues were washed with 3 l of water. The combination of wash water and filtrate gave 51 l of a clear liquid. This was sent at a flow rate of 2 through a resin column which was charged with 2.5 l of a decolorizing resin and which was about V20 of the volume of the liquid to be treated. The elution was over after 10 hours. The column was washed with 5 liters of water, and washing water and eluate were combined to obtain 55 liters of liquid. Sodium carbonate and sodium hydroxide were added to this eluate to adjust the pH to 5.5 and concentrated in vacuo at 50 to 55 ^: C. to obtain 3.78 kg of a seasoning paste which had excellent taste-improving properties and the following composition:

Composition of the wort (in weight)

Water 12.5 ⁰ zo

Total nitrogen 10.5 ⁰ zo

Alanine 10.3 ⁰ zo

Lysine 2.0 ⁰ zo

Leucine 3.7 »Zo

Thiamine 4.3 - / Zg

Pantothenic acid 37.1 γ / g

Pyridoxine 1.5 γ / g

Folinic acid 1.8 γ / g

various other amino acids were also included

NaCl 12, S ⁰ Zo

Glutamic acid 30.1 ⁰ zo

Aspartic acid 10.1 ⁰ zo

Valine 4.0 ⁰ zo

Riboflavin 16.4 γ / g

Nicotinic acid 66.6 γ / g

Inositol 375.0 γ / g

Example 4

1001 of a broth which was obtained by fermenting glutamic acid and which in 100 ml of 1.2 kg of glutamic acid, 0.5 g of alanine, 0.5 g of aspartic acid, 0.3 g of glutamine, 0.1 g of lysine, 0.1.5 g of valine 0.13 g of leucine, 1.4 g of microbes (measured in the dried state) and peptides, proteins, vitamins, sugars and the like were added with sodium carbonate to adjust the concentration to 0.03 η. It was then concentrated in vacuo at 50 ° C., and 50 l of the concentrated liquid was obtained. Hydrochloric acid was added to this liquid in order to adjust the pH to 7.2, and then it was heated ^{to 105 ° C. for 10 minutes.} After cooling, 1.8 g of a protease were added to the solution and decomposed with stirring in 16 hours at 50 to 55 ° C. The hydrolyzate was adjusted to a pH of 4.5 with hydrochloric acid and filtered. (70.2 ⁰ Zo water content) 5 1 2.5 1 filtrate and undecomposed residue was obtained. To the undecomposed residue, 1.3 kg of concentrated hydrochloric acid (35 ⁰ Zo) was added, and then the

7 ^ 8

Residue by heating for 16 hours at 105 ° C by treatment with the decolorizing resin crlial-

hydrolysicrt, whereby a hydrolyzate was obtained, was combined. The combined liquid

in which the ratio of total nitrogen to was adjusted with caustic soda to a pH of 6.2

AminostickstolT was 2.1. set, concentrated in vacuo at 45 to 50 ° C,

2 I 5 were added to the hydrolyzed liquid and 2.1 kg of wort in paste form were obtained. the Water and filtered this, in addition to the wort obtained containing sodium-5'-inosinic acid and Filtrate obtained a residue. The latter was made with amino acids such as glutamic acid, peptides, vitamins 2! Water washed. Washing water and filtrate and the like, and had a meat-like taste. Awarded 7 1 fluid which under reduced various tests including a functional Pressure were concentrated to 1.5 liters. The concentrated tests showed that the wort was grueled excellently Solution was with the liquid, which had taste-improving properties, enzymatic treatment was obtained, combined The following table shows the analytical Ergc-b- and with sodium hydroxide solution to a pH value of 6.4 in the wort of this example: placed. This liquid was then concentrated under reduced pressure at 45 to 50 ° C. Analysis of the wort and received a seasoning in paste form. The received

Product had excellent taste-water 13.7 ¹ Vo

Sending properties and contained various NaCl 10.6%

Amino acids, peptides, organic acids, vitamins thiamine 10.0 γ / g

and the like and was suitable for all seasoning purposes. 20 pantothenic acid 40.0 γ / g

Pyridoxine 3.0 y / g

Example 5 Folinic acid 3.0 γ / g

Total nitrogen 7.7%

100 1 of a fermentation broth of 5'-inosinic acid, which is replaced by 5'-inosinic acid 6.1%

Cultivate Brevibacterium ammonlagees KY »5 riboflavin 31.0 γ / g

3454 was obtained in the usual way, nicotinic acid was 0.2 γ / g

adjusted to a pH of 4.5 and resulted in biotin 0.03 γ / g

centrifugation 8.4 kg cell mass (water content, inositol 540.0 γ / g

70.2 ⁰ M. To obtain a cell milk one added

this 52 liters of aqueous 0.03 η sodium hydroxide solution were added. This 3p This seasoning can be used alone, if you wish, with the Cell milk was processed for 1 hour with stirring at 60 ° C. HCl hydrolyzate of the insoluble function heated in order to become the cell walls and membranes, which after the dissolving according to the invention, then aqueous 2 η-hydrochloric acid was added, drive can be prepared, to adjust the pH to 7.0, and then heated to 105 ° C for 10 minutes. After the 35 B e i s ρ i e 1 6 cool to 45 ° C was added to this liquid

7.7 g Streptomyces protease are added to 100 l of a fermentation broth in 10 hours, which is obtained by culturing

to decompose the cell protein at 45 to 47 ° C. Micrococcus glutamicus A. T. C. C. 13761 in usual

At the end of the decomposition, an aqueous 2 η salt manner was obtained was adjusted to a pH of

acid added to adjust the pH to 4.5 and yielded after centrifugation

put, and filtered. 50 l of filtrate and 4 kg of 5.5 kg were obtained. Microbial mass (water content 73.5 ⁰ Zo).

undecomposed residue (water content 70.2%). To obtain a cell milk, 35 liters of water were added

The filtrate was sent through a resin column containing 0.05 sodium carbonate solution. This cell

1.7 l of a decolorizing resin in its Cl form milk was heated to 55 ° C. for 2 hours with stirring

was charged, heated at a flow rate of 45. Then you put with aqueous 2N salt

from 2 through. The elution was acidic and the pH of the liquid to 7.2 a and after 15 hours

completed. The column was then heated to 105 ° C. for 10 minutes with water

washed and this wash water to the eluate cooling to 55 ° C, the liquid was added

joined. This resulted in 531 liquid. with 7.5 g of a protease, i.e. H. with l ° / o related to aul

The undecomposed residue in amounts of 4 kg 50 the total protein content, and hydrolyzed for 16 hours

was treated with 1.65 kg of concentrated hydrochloric acid (35%) at 50 to 55 ° C. The hydrolyzate contained 2.7 g /

added and hydrolyzed at 100 ° C for 13 hours. The total nitrogen and 2.2 g / l glutamic acid. Ii

Accruing hydrolyzed liquid contained in addition to this liquid were with the formation of amino

Amino acids large amounts of peptides. The acids and peptides break down 82% of the cell protein

the ratio of total N-3 to n-amino-N was 2.6. The 55 Because the hydrolyzate had a bitter taste

21 water were added to the hydrolyzed liquid to remove the bitter taste and de

added, cooled, and the following procedure was obtained after filtering coloring substances

a filtrate and a residue. The latter was made with The liquid was made by adding aqueous

21 water were added and after filtration gave 2N hydrochloric acid adjusted to a pH of 4.8

81 filtrate which was concentrated to 21. The 60 and for the purpose of obtaining a filtrate and an undecomposed

concentrated filtrate was filtered to adjust the tenth portion. The last one was washed with 21 Wassei

pH to 4.0 with aqueous 2 η sodium hydroxide solution. The filtrate and wash water gave 36.51 of a vei

puts; then you filled the filtrate with water, clear liquid, which then through a

on until a liquid amount of 5 1 obtained resin column containing 1.81 of a decolorizing resin

and treated this with 60 g of activated charcoal, packed to 65 in its Cl form, in V 50's volume

the coloring substances and bitter substances to be removed from the liquid to be decomposed, with a current;

distant. The liquid was filtered and obtained 4.51 speed of 2.0. After sth

Filtrate. which with the aforementioned eluate, which was 10 hours the elution ended. One wusc

the resin with 4 l of water, combined eluate and wash water and obtained 50 l of liquid which was free from bitter-cream 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 r} :) to 60 ° C., and 780 g of a pasty substance which had excellent taste-improving properties and the following composition were obtained had:

Analytical results of the seasoning paste

Water 12.3%>

Glutamic acid 11.8%>

Thiamine 7.2 yig

Pantothenic acid 62.0 yig

Pyridoxine 3.2 yig

Folinic acid 3.1 y / g

NaCl 11.5%

Total nitrogen 12.3 per cent

Riboflavin 27.3 yig

Nicotinic acid 112.0% / g

Biotin 0.01 yig

Inositol 625.0 yig

Example 7

I he obtained in the same manner as described in Example 1 in an amount of 5.0 kg (water content 72.5 ⁰ Io), was mixed with 2.0 kg of concentrated hydrochloric acid ^ 35 'Vn) undecomposed part. The concentration of hydrochloric acid in the mixture was about 3.5%. It was hydrolyzed for 14 hours at 110 ° C. and a decomposed liquid was obtained with about 2.0 APL value, the ratio of total nitrogen to amino nitrogen, which proved that the liquid contained large amounts of lower peptides and Contained amino acids. After cooling, this liquid 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 in vacuo to 5 liters. To adjust the pH to 4.5, sodium carbonate and sodium hydroxide were added to the concentrate, and this was made up to 10 1 liquid with water. The liquid was then mixed with 100 g of activated carbon and stirred to remove the

ao impurities such as coloring matter and bitter taste for 1 hour. After the Filter 9.51 filtrate. 101 of the obtained filtrate were combined with 5 liters of the decomposed liquid in a manner similar to that described in Example 3. The Ver-

The combined liquids were prepared according to the procedure in Example 1 treated manner and yielded 2.2 ki of a combined wort in paste form, soft by various tests including taste directory test as excellent taste vcrbc;

sender agent was found.

2694

Claims (2)

Patent claims: 1. Process for the production of wort from the Miroben masses of the fermentation broth, which in the Obtaining amino acids or nucleic acid corresponding compounds by fermentation incurred, characterized in that the aqueous liquid that consists of bacteria contains existing microbial mass, an alkali salt of a weak acid and / or an alkali hydroxide the liquid is supplied to adjust a concentration of 0.02 to 0.1 η Heated for 1 to 3 hours with stirring to 50 to 80 ° C, the pH value then to 6.0 to 8.0 set, the liquid is heated to 80 to 130 ° C for 3 to 10 minutes, after cooling with protease added and held at 50 to 60 ° C for 6 to 48 hours to break down the protein, after After cooling, the pH is adjusted to 4.5 to 4.8, the liquid is filtered and the residue is washed out with water, from the combined wash water and filtrate using a Resin column or with activated carbon the bitter substances and colorings are removed, the eluate or The filtrate is adjusted to a pH of 5.5 to 6.5 and concentrated at 45 to 60 ° C. in vacuo will. 2. The method according to claim 1, characterized in that the remaining after filtering Residue for several hours with concentrated hydrochloric acid (35%) up to a ratio being hydrolyzed from total nitrogen to amino nitrogen by about 2, the hydrolyzate diluted with water, filtered, the residue with Water is washed, wash water and filtrate are concentrated, this concentrate with the combined solution obtained in the enzymatic treatment and this mixture after adjustment is concentrated to a pH of 5.5 to 6.5 in vacuo at 45 to 60 ° C.