DE1203221B - Process for the separation of insoluble substances from fermentation liquid containing l-glutamic acid - Google Patents

Description

FEDERAL REPUBLIC OF GERMANY

GERMAN

PATENT OFFICE

EDITORIAL

Int. α .:

C12d

German class: 6b- 16/02

Number: 1203 221

File number: A 41092IV a / 6 b

Filing date: September 5, 1962

Opening day: October 21, 1965

The invention relates to a method for separating insoluble substances from 1-glutamic acid-containing substances Fermentation liquid such as is obtained in a fermentation or fermentation process. In particular bacterial cells and colloidal substances such as proteins, that are contained in the fermentation broth, separate.

It is a process for the separation of 1-glutamic acid known from fermentation solution in which the separated from the hydrolyzed mother liquor Humic substances are used as filtration media and 1-glutamic acid crystals are obtained from the filtered solution of 97% purity can be obtained. However, this method has the disadvantage that it is only applicable to undenatured cells and that the humic substances are complex, substances which vary in their composition from case to case.

With regard to the degree of purity of the 1-glutamic acid crystals, the process according to the invention results in what will be demonstrated in more detail by exemplary embodiments, the same results. In contrast to the known method, which is only applicable to non-denatured cells, the invention can Procedures can also be used in the event of abnormal fermentation, d. H. when the bacterial cells have already decomposed and dissolved. This allows the area of application to be increased considerably, because the method according to the invention can in principle be applied to all fermentation broths, and there is no need more to check whether there is normal fermentation or not. Furthermore, in the known Process to be expected that it can not always be carried out in a precisely reproducible manner, since it The humic substances used vary in their composition from case to case varying substances. The method according to the invention avoids this disadvantage in that a chemically precisely defined compound is used.

The inventive method for the separation of insoluble matter from 1-glutamic acid-containing fermentation liquor with adjustment of the crude fermentation broth to a pH of 6, and heating the same to 60 to 8O ⁰ C with subsequent centrifugation is characterized in that the centrifuged liquid from the compounds Polyoxyäthylenalkylamine and polyoxyethylene lauryl ether with a hydrophilyophobic balance greater than 15 adds at least one, the fermentation liquid is heated and filtered off in a manner known per se. Poly processes for separating insoluble substances from 1-glutamic acid-containing substances are preferred
Fermentation liquid

Applicant:

Asahi Kasei Kogyo Kabushiki Kaisha,

Osaka (Japan)

Representative:

Dipl.-Phys. G. Liedl, patent attorney,

Munich 22, Steinsdorfstr. 22nd

Named as inventor:

Sotoo Yamamoto,

Masatoshi Nakayama, Miyazaki-ken (Japan)

oxyethylene lauryl ether with 10 to 30 oxyethylene molecules and as polyoxyethylene alkylamines, polyoxyethylene stearylamines with 10 to 30 oxyethylene molecules or polyoxyethylene oleamine with 5 to 15 oxyethylene molecules is used.

Generally contains a! -Glutamic acid containing Fermentation broth bacterial cells and organic, colloidal substances, e.g. colloidal proteins, which mainly result from a fermentation bacterium. A purification of such a fermentation broth is due to the extremely fine bacterial cells and the materials suspended in them difficult. As a result, it is almost impossible to obtain a transparent centrifugate from such a fermentation broth to win, even if in a conventional Sharples centrifuge centrifuged or filtered with an ultrafilter in which a large amount of filter substance such as Diatomaceous earth (kieselguhr) is included. The purity of the 1-glutamic acid crystals obtained from a Fermentation broth, as mentioned above, can be crystallized out, is only about 80 to 85% · In particular, it is very difficult to use such a general filtering process to produce high-purity 1-glutamic acid crystals on an industrial scale. From the above Reasons it turned out to be to remove these colloidal ones Substances that arise in the fermentation broth during the fermentation process, as required, for example the Proteins by adding hydrochloric acid into amino acids

509 718/83

3 4

to implement and the fermentation broth thereby on an increased slope with the examples given of the other

To warm up temperature. Explanation of the invention.

During the course of the invention, various processes were used. A crude fermentation broth containing 1-glutamic acid drive to the separation of bacterial cells and is neutralized with hydrochloric acid to a pH of 6 and

colloidal proteins from a raw fermentation broth under-5 heated ^{to 70 0 C for 30 minutes.} A great addiction. In particular, the investigation extended part of the bacterial cells is then centrifuged down using a Sharples centrifuge, taking over all types of surface-active compounds,

and the effect of adding surface-active substances, resulting in a centrifugate with a turbidity of about

Compounds on the filtering ability of the broth. It gives 650 °. 0.05% of poly-

it was found that certain specific upper oxyäthylenlauryläther (Emuigen 147, manufactured by surface-active compounds particularly effective KAO Soap Co.) with a hydrophilic-lyophobic

are; only a few surface-active compound balance of 16.5 was added and the mixture

fertilize, as they are described below, heated ^{to 6O 0 C for 30 minutes.} Fall on it

are extremely effective when beet molasses are organic, colloidal substances, which are bacterial

are used as sugar sources or when i ₅ cells and defoamers are in suspension

bacterial cells become abnormal and lead to a transparent, protruding

Dissolve fermentation, as in Fermentation Solution. After adding 1 percent by weight

can occur. Diatomaceous earth and subsequent pressure filtration

The effective compounds are polyoxyethylene using an ultrafilter results in

alkylamines and polyoxyethylene lauryl ethers with a 20 short time a transparent filtrate. The resulting

hydrophilic-lyophobic equilibrium greater than 15. The filtrate is a transparent solution with a turbidity

The reason why only those mentioned above are less than 10 °. There are no fixed in this solution

contain more surface-active compounds among countless types of suspension substances. (By analogy with the

of surface-active compounds are effective, turbidity of water, the turbidity is generally like that

is theoretically not exactly known. However, it is over-defined that the turbidity is 1 ° when one millimeter

surprising that only when adding the above gram acid clay is mixed with water.)

surface-active compounds and subsequent er- if in contrast to that described above

warming of the centrifuged fermentation liquid, procedure the same centrifugate with a turbidity of

which previously adjusted to pH 6 to 8 and to 60 to 650 °, from which a large part of the bacterial cells

8O ⁰ C was heated, the proteins from bacterial 30 was removed with the help of a Sharples centrifuge,

Is cells or fats from defoamers to-absorptive for 30 minutes at 6O ⁰ C heated without

be brought together so that it is added to its hydrophilic polyoxyethylene lauryl ether

Reduce properties and they take place the filtration only very slowly by coagulation, even if

can be caused to precipitate. In addition, the filtration using diatomaceous earth

from almost all are cationic, anionic and non-35 is carried out, and indeed in large quantities of

ionic surface-active compounds far less than 5 percent by weight. The resulting filtrate has

ger than the mentioned surface-active connec- tion of the suspended organic, colloidal

fertilize effectively. It is also surprising that the substance has a turbidity of 500 °.
Comparison with Polyoxyäthylenoleiläther, Polyoxyäthy-

Lencethylether, polyoxyoctylphenyläther and the like
Compounds very similar to polyoxyethylene lauryl ether

seem to be merely polyoxyethylene lauryl ether, a crude fermentation broth containing 1-glutamic acid

which shows extremely pronounced effectiveness. was reduced to a pH of 6 with hydrochloric acid.

Siert of those used according to the invention; it was for one hour at 70 ⁰ C eroberflächenaktiven compounds are polyoxyethylene 45 wäimt; Then, using an alkylamine, the centrifugation was somewhat more effective than the polyoxyethylene Sharples centrifuge, and the centrifugate was lauryl ether, in the case where the beet was deposited. 125 g of polyoxyethylene lauryl ether with molasses are used as sugar sources, 13 oxyethylene molecules of Emuigen 120, manufactured by or in the case of abnormal fermentation on KAO Soap Co., occurred in 2501 of the centrifugate due to bacteriophages. In this anomalous 50 brought. The centrifugate had a turbidity of 650 °. len fermentation dissolve the bacterial cells and the resulting mixture was filtered to in solid colloidal substances which th to 70 ⁰ C and then heated under difficult overall Verdie separation of the bacterial cells turn an ultrafilter for 20 Minuverwandeln; the ultrafilter, even when using a Sharples centrifuge, had a filter area of 1 m ² ; the filtration was carried out so that the resulting broth had a turbidity of 55 under a pressure of 1 to 2 kg / cm ² . Has more than 5000 ° after 20 minutes. The filtration was finished. The cloudiness of the fiI

As examples of the trates which can be used according to the invention, it was 20 °. On the other hand, if the same filtrate

Surface-active compounds are polyoxyethylene with a turbidity of 650 ° under the same conditions

oleilamin with 5 or 15 oxyethylene molecules and gungen was filtered, but no polyoxy-

Polyoxyäthylenlauryläther with 13 or 25 Oxyäthylen- 60 äthylenlauryläther was added, the was for the

to name molecules. Furthermore, the filtration time in particular is 2 hours. The cloudiness

effective following compounds: Polyoxyethylene- this filtrate was at 400 °.
stearylamine with 10 to 30 oxyethylene molecules or

Polyoxyäthylenoleilamin with 5 to 15 Oxyäthylenmole- Example 2

kulen or polyoxyethylene lauryl ether with 10 to 65

30 oxyethylene molecules. In a raw fermentation broth containing 1-glutamic acid,

The following description of preferred compounds is a particularly solid emulsion of bacterial

Execution of the invention is used in the confluence of cells, as occurs in an abnormal fermentation,

I 203

Hydrochloric acid was introduced so that the pH was adjusted to 6. The resultant broth was heated for 30 minutes at 70 ⁰ C and then centrifuged with a Sharples centrifuge, so that a centrifugate gave a turbidity of 5000 °. 125 g of polyoxyethylene alkylamine with a hydrophilic-lyophobic balance of 16.5 (Amiete 310, manufactured by KAO Soap Co.) were introduced into 250 liters of the centrifugate obtained in this way. The resulting mixture was heated ^{to 70 ° C. for 20 minutes.} After adding 1.2 kg of diatomaceous earth, filtration was carried out under a pressure of 1 to 2 kg / cm ² with the aid of an ultrafilter, which had a filter area of 1 m ² . The filtration was finished after 20 minutes. A transparent filtrate with a turbidity of 30 ° resulted. On the other hand, when the same centrifugate with the turbidity of 5000 ° was filtered under the same conditions except that no polyoxyethylene alkylamine was added, the filtration ao was not finished even after 2 hours. In addition, the filtrate was a cloudy solution with a turbidity of 3000 °. Thus, polyoxyethylene alkylamine is very effective in cleaning a fermentation broth which has undergone abnormal fermentation.

Claims (3)

Patent claims: 1. Process for the separation of insoluble materials from? -Glutamic acid-containing fermentation liquor with adjustment of the crude fermentation broth to a pH of 6, and heating the same to 60 to 8O ⁰ C, followed by centrifugation, characterized in that the centrifuged liquid from the compounds Polyoxyäthylenalkylamine and adding at least one polyoxyethylene lauryl ether with a hydrophilic-lyophobic balance greater than 15, heating the fermentation liquid and filtering it off in a manner known per se. 2. The method according to claim 1, characterized in that the polyoxyethylene alkylamines Polyoxyethylene stearylamine with 10 to 30 oxyethylene molecules or polyoxyethylene oleamine used with 5 to 15 oxyethylene molecules. 3. The method according to claim 1, characterized in that there is a polyoxyethylene lauryl ether used with 10 to 30 oxyethylene molecules. Considered publications:
British Patent No. 869,948. 509 718/83 10.65 © Bundesdruckerei Berlin