CS197373B1 - Method of preparation of the technical preparation tryptophane particularly for the fodder purposes - Google Patents

Description

The present invention relates to a process for the preparation of a tryptophan preparation, particularly for feeding purposes, which is stable, non-hygroscopic and suitable for the preparation of feed premlxes.

As is known, the grains, as the main feed component of monogastric livestock, are composed of their proteins defioient to L-tryptophan (beside L-lysine). Accordingly, there is a growing need to add this amino acid to compound feeds and, in parallel, the need for an inexpensive yet qualitatively satisfactory technical preparation.

More recent methods for obtaining L-tryptophan are based on biosynthesis. The fermented broth after culturing a suitable microorganism is processed in several stages to produce a high purity preparation. These procedures are complex and expensive, the product obtained is expensive and more suitable for medical use.

Methods for the preparation of pure tryptophan are well known in the art for various operations, such as metal salt precipitation, ion exchange adsorption, activated carbon and the like, extraction, molecular filters, etc. about 50% of fermented soil 99.8% product. After adsorpol, the accompanying amino acids are eliminated by partial elution of the salt solution, the eluate is purified, crystallized and recrystallized. The most commonly used adsorption for strongly acidic cation exchangers or a combination of two different ion exchangers is obtained with water-alcohol-ammonia-based eluates, from which a crystalline product is obtained after purification (French Patent Specification No. 2 000 641, US Patent No. 154425J). No. B. Beker et al., Ex. Bioohim. Microbiol. 7, 103).

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In an effort to obtain in a simple and inexpensive manner tryptophan in the form of a technical preparation suitable for feed purposes, a process for the preparation of such a preparation from fermented broth after the cultivation of the production microorganism has been developed. SUMMARY OF THE INVENTION The soil is inactivated by acidification to a pH of 1.2 to 4.0, preferably 2.0, with the addition of a non-allergenic or cationic surfactant and heating to a temperature of 50 to 90 ° C, preferably 55 to 65 ° C, for 3 to 10 minutes, then cooled to 0 to 10 ° C, preferably 4 to 6 ° C, filtered, the filtrate contacted with a sulfonated styrene-divinylbenzene copolymer in H- an octyl, from which tryptophaa is eluted with an aqueous ammonia solution in a concentration of 0.5 to 1 N, at a temperature of 35 to 45 ° C, the eluate is concentrated at a temperature of not more than 40 ° 0 to a dry matter content of 15 to 20%; for example in a spray drier.

Sulfuric, phosphoric, oxalic or triohloroic acids may be suitably used for the initial acidification of the soil, individually or in a suitable combination.

The described special pretreatment of fermented broths, ie acidification, wetting and short-term heating, will facilitate and accelerate the removal of soluble high molecular weight ballasts, mainly proteins, substantially facilitating the subsequent adsorption and elution of tryptophan from the cation exchange resin. An eluate of a qualitatively suitable quality for direct drying in a spray drier is obtained, whereby a preparation with a relatively high content of active substance, very hygroscopic, stable under normal conditions of storage, suitable for processing into premixtures for compound feed, is obtained. Otherwise, without spelar soil treatment, the product is almost non-compliant, hygroscopic, tacky, unstable, as shown in the comparative embodiment.

4. Polyethylene glycol sorbitan monooleate surfactants and cationic alkylarylammonium halide-based quaternary salt surfactants have proven to be suitable for the removal of soluble ballast substances from the fermented soil.

Further details of the process according to the invention are apparent from the examples which illustrate the process but do not limit it in any way.

Example 1

To 10 liters of fermented broth containing 10.7 g of tryptophan per liter, 1% (w / v) of oxalic acid and sulfuric acid were added immediately after reaching the maximum of the product to a pH of 2.1, and 3% (w / v) of non-halogenated wetting agents (polyethylene glycol sorbitan monooleate). The treated soil was quickly heated to 55 ° C with a 5 minute delay, then cooled rapidly to 5 ° C and filtered through diatomaceous earth. The filter residue was washed with 1 liter of water. 9.5 liters of a filtrate containing 9.2 g / liter of tryptophan were obtained, totaling 87.4 tryptophan, i.e. a yield of 61.7% of theory.

The filtrate was acidified with sulfuric acid to pH 1.3 and passed to a column at 20 ° C

0.5 liter of cation exchange resin in H-loop (and 2% by weight of divinylbenzene) at a rate of 1 liter / hour. X penetration θ * 4 g / liter tryptophan occurred after 3 * 5 liter filtrate flow, saturation after 9 * 5 liter flow.

The cation exchanger was then washed with 0.5 L of 50 ° C warm water to obtain 6.0 L of a conoentraoi effluent of 4.7 g (i.e. 26.2 g of tryptophan).

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The cation exchanger was eluted with 1.5 L of 0.6 N ammonia at 40 ° C, with a rate of 0.5 L / h.

650 ml of a rich eluate (fraction with a content greater than 10 g / liter of tryptophan) of 78 g / liter (i.e., 50.8 g of tryptophan) was obtained. The remaining poor eluates contained 4.2 g of tryptophan. A resin capacity of 110 g / liter, an elution yield of 92.4% and an adsorption yield of 92.9% were achieved.

The bluate was concentrated at a temperature of not more than 38 ° C to a dry matter content of 17% by weight to recover some of the ammonia used. The concentrate was then dried in a spray drier at 125/70 ° C. 74 g of a technical preparation with a content of 65.8% tryptophan (3% moisture), i.e. 48.7 g of pure tryptophan, were obtained. Concentration and drying yield was 95.9% · The stability of the preparation after 6 months of storage in a sealed container under normal conditions was better than 95% · The preparation was only slightly hygroscopic, at 87% relative humidity the equilibrium moisture content of the preparation was 6.8% .

Toxicity of LDjq 2.92 g / kg (limits 2.64 to 3.23 g / kg).

Standard Toxicity (99%) LD 50 q 3.47 g / kg (limits 2.73 to 4.41 g / kg).

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10 liters of fermented broth containing 8.1 g of tryptophan per liter were treated according to the procedure of Example 1 except that phosphoric acid and a cationic surfactant were used instead of sulfuric acid and nonionic surfactant. After the treatment, the soil was centrifuged on a sedimentation centrifuge to obtain 7.2 liters of filtrate of 7.8 g tryptophan per liter. Further processing according to Example 1 yielded a preparation of adequate yield and quality, i.e. non-hygroscopic and stable, suitable for the preparation of feed premixes.

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The fermented broth of Example 2 was treated by acidifying 1% (w / v) of triohloroic acid, 1% (w / v) of oxalic acid and phosphoric acid to pH 2 after reaching the maximum production. The further procedure was identical to Example 1. Penetration from the column at the adsorption occurred after 75% tryptophan flow of practical column capacity, saturation was achieved after 160% tryptophan flow. Further processing as in Example 1 yielded a preparation of adequate yield and quality suitable for the production of feed premixes.

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The fermented soil according to Examples 1 or 2 was acidified with sulfuric acid or hydrochloric acid to pH 1.4 and immediately passed from below to a cation exchange column as in Example 1. The adsorption in the fluidized state under the conditions of Example 1 penetrated already after a flow rate of 20 to 28% capacity of tryptophan, to saturate after flow rate of 250 to 320% of the capacity of the column for txyptophan, which was not a good precondition for the development of the technological process. Further treatment according to Example 1 yielded 71 g of a preparation containing 52% tryptophan. At a relative humidity of 87%, the equilibrium moisture of the preparation was established at 17.0% and the preparation glued together. This procedure gave an unfavorable result against Examples 1-3.

Claims (2)

SUBJECT OF THE INVENTION Process for the preparation of a technical preparation of tryptophan, in particular for feeding purposes, from fermented broth after cessation of cultivation in a producing microorganism, characterized in that the soil is inactivated by acidification to a pH of 1.2 to 4.0, preferably 2, 0, with the addition of a non-ionic or catlon-active surfactant and heating to a temperature 50 to 90 ° C, preferably 55 to 65 ° 0, for 3 to 10 minutes, then cooled to 0 to 10 ° 0, preferably 4 to 6 ° C, filtered, the filtrate is contacted with sulfonated styrene-divinylbenzene copolymer in H-loop, from which tryptophan is eluted with an aqueous ammonia solution in conoentraol of 0.5 to 1 N, at a temperature of 35 to 45 ° 0, the eluate is concentrated at a maximum of 40 ° C to a dry matter content of 15 to 20%, after which the concentrate is dehydrated, for example in a spray drier. 2. Process according to claim 1, characterized in that sulfuric, phosphoric, oxalic or triohloroic acids are used for acidification, individually or in a suitable combination.