HU192549B - Process for producing protein concentratum free from saponin from lucerne - Google Patents

Abstract

The present invention relates to fodder for livestock based on lucerne (alfalfa). This fodder is characterised in that microbiological methods are used to remove from it, totally or partially, anti-nutritional substances, firstly saponins, sapogenins, sapogenols, medicagenic acid or their derivatives. Application to the feeding of monogastric animals.

Description

BACKGROUND OF THE INVENTION The present invention relates to the production of a product containing at least 45% protein, free of antinutritive substances, in particular saponins, obtained by fermentation from green parts of alfalfa. The product of the present invention is capable of adjusting the protein levels of various feeds. The resulting product may be mixed with other feed components in the form of juice, concentrate or dried composition according to the purpose of storage and technology.

It is known that monogastric animals may not or only to a limited extent be fed lucerne or alfalfa press juice, sticks, dried preparations, protein concentrates due to the adverse physiological effects of antinutritive substances in lucerne, in particular saponins. FT. Szabó Mária: Antinutritive factors that reduce the biological value of plant nutrients. Animal Husbandry and Animal Feeding, 31 (3), 273-287 (1982)]

The high saponin content of alfalfa concentrates (5-12 g / d.kg) in monogastric stomachs - e.g. in poultry and pigs - inhibits growth, reduces weight gain, adversely affects poultry egg lysates.

Saponins and saponin-like compounds may exert the following physiological effects:

1. hemolysis of erythrocytes;

2. undesirable lowering of blood and liver cholesterol;

3. inhibition of growth;

4. inhibition of smooth muscle contraction;

5. inactivation of digestive enzymes;

6. inhibition of gastrointestinal absorption.

Cheeke, P.R .: Nutritional and physiological implications of saponins. A review. Can. J. Anim. Sci. 51. 621–632. (1972).

Because of their adverse physiological effects, dried fodder products made from lucerne have been displaced from poultry feeds. The production of protein concentrates from lucerne has been discontinued in many places, and the limited production is essentially about the production of carotene-chlorophyll dye concentrate. The methods of concentrating the protein content of alfalfa by physical methods also concentrate the saponins in the final product. Thus, these formulations are not included in the PROXAN (USA), PX (French), VEPEX (Hungarian) feed formulations to replace the amount of protein required (Kállai Kraloványszky: Biology of Feeding. : Certification of Feed Proteins, Agricultural Publisher, Budapest, 1981, 208-210.

Processes for making said and similar products do not accomplish the object of the present invention.

It is an object of the present invention to provide a low saponine product (0 to 1.0 g / kg) containing at least 45% protein from alfalfa which is suitable for feeding to at least 10% of monogastric animals.

The present invention is based on the discovery that the saponin content of lucerne phytonutrients by fermentation is mono yellowish brown. The mature aerial mycelium is a powdery, light yellowish-green, later yellowish-gray. Does not form pigment.

Glycerol Asparagine Agar (ISP-5): The vegetative mycelium is light tan. The mature aerial mycelium is cream in color. It forms a spore. It does not produce pigment.

Yeast Line Malate Extract Glucose Agar (ISP - 2): The vegetative mycelium is tan. The aerial mycelium is abundant, initially white, the mature sporulated aerial mycelium is yellow-brown. It forms a spore. It does not produce pigment.

Peptone Yeast Extract Iron Agar (ISP-6): The vegetative mycelium is tan. It produces a yellow-brown pigment.

Tyrosine Agar: Produces a yellow-brown pigment.

The strain reduces nitrate and does not produce hydrogen sulphide.

Biocmitic activity

D-Glucose (+) Glycine (+) D-Fructose (+) Beta-Alanine (+) Maltose (+) DL-alpha-alanine (+) Mannis (+) DL Selector (+) Xylose (-) DL-lcucin (+) L-rhamnose (-) DL-nor-leucine (+) Rallynose (-) l-aspartic acid (+) Arabinose (-) L-Asparagine (+) Lactose ( -) L-glutamic acid (+) Dulcit (-) D-arginine (+) Inositol (-) DL-lysine (+) Cellulose (-) By DL (+) Sucrose. (4 -) DL-lriptophane (+) Starch (+ -) L-Histidine (+) L-Cystine (+) DL-Methionine {+) (NH ₄ ) ₂ HPO ₄ (+) Urea (+) Peptone (+) Dl-Isoleucine (Uncertain) L-Liroz.in (-) NaNO ₂ (-)

Does not increase at 'C'.

It does not grow in a microaerophilic manner. It is capable of breaking down steroid compounds. Optimal growth: ISP 2 bog ,, 28 30 'C.

Based on the above, the strain can be classified into the species group Str. Griseus.

Streptomyces sp. GATE No. 38 strain inoculum by shaking submerged culture with 0.5-5.0% w / v water soluble carbohydrate, preferably

1.5% starch as the sole source of carbon, a source of organic biofuels, preferably corn jam, an inorganic nitrogen source and inorganic salts. pH 5.0-8.0, preferably 6.5; culture temperature: 20-30 'C, preferably 27' C; incubation time: 48 to 120 hours, preferably 96 hours.

The resulting inoculum (1 to 10% v / v, preferably 5% v / v) is used to inoculate 0.5 to 5.0% water-soluble carbohydrate, preferably 2.0%, of the fermentation broth made from saponin-lucerne lucerna presle. starch or equivalent industrial maize flour, inorganic nitrogen and inorganic salts. Aerobic Submerge. fermentation is carried out for 70 to ISO, preferably 120 hours. Other conditions for culturing are the same as for inoculum preparation.

The resulting fermentation broth is heat-treated at 70-100 ° C, preferably 90 ° C, to precipitate the protein, filtered (92,549), and the filtrate is concentrated by evaporation. The biomass thus obtained is dried in a known manner.

The product containing 45-55% of protein is utilized in feeds, preferably in the feed of our monogastric stomachs, instead of 10-50% of the protein portion of the formulations, preferably soy.

It is also possible to use concentrated fermentation broth in wet feeding technologies, which saves energy and technological time of drying.

In our experience, saponification of lucerne juice juice can also be accomplished using a short cycle semi-continuous fermentable feed yeast (Candida utilis GATE G-1, deposited on January 14, 1986 in the OKI National Collection under MNG 00341).

Λ strain description:

Morphology: vegetative cells prolate multilaterally, cells ovate, cylindrical, 7-13x3-5 pm. The pseudomycelium consists of undeveloped, longer cylindrical spore-forming cells, which produce smaller blastospores. It does not form arthrospores, does not produce ascospores, basidiospores.

Columns: round, prominent, d = 4-5 mm, silky light, light cream in color.

Biochemical activity

Nitrate assimilates Inulin assimilates Eritrit not assimilates Galactose (-) (asc) Sucrose (+) (fermentation) Maltose (+) (asc) Lactose (-)

Raffinose (-)

Glucose (+) (crj.) Galactose (-)

Maltose (-) (inc.)

Lactose (-) (excl.) Raffinose (-) (inc.) For hair (3) (asc) L-arabinose (+) (asc) D-arabinose (+) (asc)

Inositol (-) (ass.)

Inulin (3 -) (Ass) Starch (+ -) (Ass) Saponin (+) (Ass)

Optimal growth

Wickerham's yeast extract - malt extract - peptone - glucose - agar., PH 5.0, 28 'C.

Staining: lactophenol aniline, malachite green safranin. Based on the above, the strain can be defined as Candida utilis.

Candida utilis GATE G-1 strain is prepared by submerged culture inoculum, 1.0-10% w / v of water-soluble carbohydrate, preferably 8.0% molasses, non-nitrate inorganic nitrogen source, preferably diammonium hydrogen phosphate, preferably organic dried material. medium containing feed yeast and inorganic salts, pH 4.0-7.0, preferably 5.0; breeding temperature. 20-35 'C, preferably 28 ° C; fermentation time: 30 to 72, preferably 48 hours.

The inoculum prepared as above (5-20, preferably 10% v / v) is inoculated from the saponin-containing lucerne press or diluted lucerne press, 1.0 to 10.0% w / v of a water-soluble carbohydrate, preferably molasses, a bioavailable source. ₅ calories, a medium containing inorganic nitrogen sources other than nitrate, preferably diammonium hydrogen phosphate and inorganic salts. pH: 4.0 to 7.0, preferably 5.0; culture temperature: 20 to 35 ° C, preferably 28 ° C; fermentation time: 30 to 72, preferably 50 hours, or until the saponin content decreases from an initial level of 0.5 to 1.0 mg / ml to less than 0.01 mg / ml. Alternatively, the semi-batch fermentation was carried out so that explained 80% of the finished product of fermentation described processing, the residue ₁₅ was used as inoculum to start the next cycle ráfejtve fresh medium. The resulting fermentation broth is heated to 70-100 ° C, preferably 90 ° C, to precipitate the lichen's milk, the precipitated protein is filtered off, concentrated, and the biomass obtained is dried in a known manner. The 45-55% protein-containing product is used as a protein source in feeds, preferably 10-50% of the soybean content in single-gastric feeds.

Alternatively, concentrated fermentation broth can be used as a protein source in wet feed technology.

The press juice and concentrate can be preserved, if necessary, so that the pH is between 2.0 and 4.0,

The 2Q is preferably adjusted to 3.5 with a mineral and / or organic acid, preferably formic acid.

It has been found that the process of the present invention can also be carried out with a filamentous microscopic fungal strain (Penicillium sp., GATE GX -5 le35, dated January 14, 1986, in the OKI National Collection, MNG-00342).

Description of strain

Morphology: colonies on Czapek agar are 3-4 cm in diameter, velvety, later dusty. The mature kouidiiunos colony is initially greenish-gray, older colonies are brownish-gray. At the edge of the battery, the white rim is 2 - 3 mm wide. The substrate mycelium (colony of the colony) is yellowish white. Conidiophores are 150 to 300 µm long, 3 to 4 µm wide. Brushes have an asymmetric arrangement, sometimes of the Divaricata type. Symmetrical Monoverticillata types can also be observed. The 3-5-membered fialides are 8-10x2-3 pm, tapering to the neck. The conidia are generally spherical, smooth, 3 to 3.5 µm in diameter.

Biochemical activity

Glucose (t)

D -galactose (+) Sucrose (+) Maltose (+) L-sorbose (+) D-xylose (+) Mannitol (+) Sorbitol (3) Saponin (+)

Dulcil () Meso-inositol (-) Lactose (+ -) D-raffinose (3- -) L-rhamnose (3-) L-arabinose (-) Adonite (3-)

-3192 5 ^ 9

Optimal Growth: on Sabouraud Glucose Agar, pH 5.6, 28 'C. Staining: Air and substrate mycelium are well stained with lactophenol cotton wool.

Based on the above, the strain can be safely classified into the genus Penicillium, with moderate probability into the notatum species group.

Penicillium sp. Substrate cultures of G ATE GX-5 are inoculated with 1.0-20.0% w / v water soluble carbohydrate, preferably 3.0% w / v sucrose, 0.1-0.5% w / v inorganic nitrogen, preferably on a medium containing 0.2% (v / v) ammonium nitrate, 0.2-2.0% (v / v) organic biobased material, preferably 0.2% (v / v) malt extract and / or meat extract and inorganic salts. pH: 4.0 to 8.0, preferably 6.0, culture temperature: 20 to 37 'C, preferably 28' C, fermentation time 48 to 96 hours, preferably 72 hours. 5 to 20% (v / v) preferably 10 to 10% (v / v) inoculum of saponin-containing lucerne juice or diluted lucerne juice, 1.0 to 10.0% w / v of water-soluble carbohydrate, preferably 6.0% by weight of molasses, 0.2 A medium containing 0.3% biofuel source, preferably 0.2% corn jam, 0.1-0.5% inorganic nitrogen source, preferably 0.2% ammonium nitrate and / or ammonium phosphate, and inorganic salts. pH; 4.0 to 8.0, preferably 6.0; culture temperature: 20 to 35 'C, preferably 28' C; fermentation time: 48 to 96 hours, preferably 78 hours, until the saponin content decreases from 0.5 to 1.0 mg / ml to 0.01 mg / ml. The fermentation broth is heated to 70-100 ° C, preferably 90 ° C, and the biomass thus precipitated is filtered off, concentrated by evaporation and dried in a known manner. The final product having a protein content of 50-55% is used as a protein source in feeds, preferably in monogastric feeds, to replace the protein component, preferably soy. Alternatively, concentrated fermentation broth can be used in protein feed wet feed technology. The press juice and concentrate may be preserved, if necessary, by adjusting the pH to 2.0-4.0, preferably 3.5, with a mineral and / or organic acid, preferably formic acid.

A particular advantage of the invention is that the saponin-free, high-protein product of the present invention is suitable for complete replacement of soy in the feed of monogastric stomachs in an amount of 10-50%.

A further advantage is that the twin product formed in the process, the fibrous compression residue, is also a valuable feed and / or feed additive that can be used fresh, pressed or dried in a known manner to feed ruminants and our entire stomach.

It is a notable advantage that the process can also be used to remove saponins from dried alfalfa varieties, flours and concentrates of commercially grown high saponin (5-13 g / kg).

According to the invention, lucerne phytomass is produced by cultivation in the field with known varieties of lucerne, preferably with a variety selected for lower saponin content (1-5 g / kg). Green alfalfa is chopped, disintegrated and squeezed. The press juice is prepared from Streptomyces sp. GATE-38 (MNG 00282) or Penicillium Sp. GATE GX - 5 (MNG 00342) or Candida utilis GATE

Fermented with strains G-1 (MNG 00341) to a saponin content of 0-0.1 mg / ml as measured by hemolysis. The fermented press juice is concentrated in a known manner, preserved if necessary or dried. The products thus obtained are used in amounts determined by the variety and the rearing technologies for protein supplementation, preferably soy replacement at 10-50%.

Strain maintenance of said microorganisms can be achieved by any known method, for example, by lyophilization, immersion in sterile paraffin oil in spore state, sporulation on sterilized seeds of cereals. Preferably, inoculum preparation is carried out on cultures passaged and maintained on saponin-containing lucerne juice medium containing from 0.5 to 1.5% w / v of a water soluble carbon source solidified with 2.0 to 2.51% agar agar. In the subcultures so maintained, saponin tolerant and degradation lines are safely maintained.

The following examples illustrate the process of the invention in more detail.

Example I

Streptomyces sp. GATE-38 (MNG 00282) was selected and maintained on saponin-containing alfalfa juice agar medium. A spore suspension is prepared from the surface of a 10-day slant culture with 0.02% Tween 80 solution. An inoculum culture of the spore suspension is prepared by shaking with 5 to 5 ml of the following medium:

(NH ₄ ) ₂ SO ₄ 0.3%

KH ₂ PO ₄ 0.1%

K ₂ HPO ₄ 0.2%

MgSO ₄ .7 H ₂ O 0.1%

Corn jam (powder) 0,25% Starch 1,5%

Dosage: 100-100 ml enkcnt.

The pH is 6.8, the incubation temperature is 27 'C, and it is cultivated on a shaker at 250 rpm for 96 hours. This culture was used to inoculate the next fermentation step at 5.0 volume%. The composition of saponin fermentation broth is as follows:

0.3%

0.1%

0.2%

0.1%

0.4%

2.0%

5000 ml (0.8 mg / ml saponin tartrate with hemol. Test). 0.1% glass, feet. fermentor (KUTESZ). Mixing: 250 / pcrc, air: 6 liters / min, temperature: 27 ° C ± 1 ° C; fermentation time: 120 hours. Saponin content at the end of fermentation: 0 - 0.01 mg / θθ ml. The 4.0 L fermentation broth requested for processing is heated to 90 ° C and maintained at this temperature for 5 minutes. The heat-precipitating protein is isolated by filtration. Wet biomass: 528 g / 4.0 l, 132 g / l, 13.2% of the starting material. The wet biomass was dried at 60 g5 ° C to 181.6 g of dry fatty acid 45 (NH ₄ ) ₂ SO ₄ kh ₂ po ₄ k ₂ hpo ₄

MgSO ₄ .7H ₂ O

Corn steep liquor (powder)

Starch

Luccrnapréslé

Antifoam (sunflower oil) pH 6.8

Unit: 10 liters.

192,549 were obtained, which was 34.4% of the wet material and 4.54% of the crude juice. The crude protein content of the material was found to be 55.2%. Carotene: 222 mg / kg, xanthophil: 948 mg / kg.

Example 2

Lucerne pressed agar was selected and maintained on Penicillium sp. Gonid GX-5 (dep. MNG 00342) was prepared from a surface of a sporulated, 8-day-old culture with a sterile suspension of 0.02% Tween 80 in conidia. Prepare a suspension of 3 to 3 ml of inoculum culture medium in the following medium:

NH ₄ NO ₃ 0.20% kh _{2 after 4} 0.15% k ₂ hpo ₄ 0.20% MgSO ₄ .7 H ₂ O 0.10% saccharose 3.0% Mafátakivonat 0.20% meat Extract 0.20% Dosage 100 - Every 100 ml.

pH: 6.0, Culturing: Plate-rotary shaker at 250 rpm at 28 ° C for 72 hours. The shake inoculum culture is inoculated with 10% (v / v) saponification medium containing:

0.20%

0.15%

0.20%

6.0%

0.20%

5000 ml (0.8 mg / ml saponin Heol assay).

(NH ₄ ) ₂ HPO ₄ KH ₂ PO ₄ K ₂ HPO ₄ Molasses

Corn jam (powder) Lucerne juice

Antifoam (sunflower oil) 0.10% pH 6.0

Appliance: 10 liters, bottle, foot, fermenter (TUT). Mixing: 260 / pcrc, air: 8 liters / min, temperature: 28 'CT I' C: fermentation time: 78 hours until the haemol. the saponin content measured by the test is reduced to 0-0.01 mg / ml.

The 4.0 L fermentation broth to be processed is concentrated to 40% solids and then spray-dried. The above procedure yields 136.8 g of saponin-free product, 3.42% of the processed pcl juice, having a crude protein content of 52.6%, carotene: 220 mg / kg, xanthophil: 900 mg / kg.

Example 3

A cell suspension of a maintained Candida utilis GATE G-I (dep. No. MNG 00341) cultured on lucerne press agar was prepared in cell suspension. Prepare a shake inoculum culture for 5 to 5 ml of the cell suspension on the following medium:

(NH ₄ ) ₂ HPO ₄ 0.30%

KH, PO ₄ 0.10%

MgSO ₄ .7 H _a O 0.05%

CaCLj 0.05%

Yeast Extract 0.10%

Molasses 8.0%

Dosage: 100 to 100 ml, pH 5.0. Cultivation: Plane-rotary shaker, 250 rpm, 28 ° C, 48 hours. The shake culture is inoculated with 10% (v / v) saponin fermentation broth. It has the following composition: 0.30%

0.10%

0.20%

8.0%

5000 ml (0.8 mg / ml saponin hemol assay).

4/2 ** ' ^v 4

KH ₂ PO ₄ Corn jam (powder) Molasses

Lucerne juice

Anti-foaming agent (sunflower oil + 2.5% Tcclomctan)

0.10% pH: 5.0

Device: BIOTEC foot. fermentor, 10 liters.

Mixing: 250 / min, air: 7 liters / min, temperature: 28 ° C ± 2 ° C, fermentation time: 50 hours, until the saponin content measured with the hemol.tc is reduced to 0-0.01 mg / ml. Workup is carried out as described in Example 2. The saponin-free dry protein concentrate thus obtained was 134.0 g, which is 3.35% of the processed press juice. Crude protein content: 56.2%, kerotin: 218 mg / kg, xanthophil: 895 mg / kg.

Example 4

The procedure for inoculum preparation and fermentation of saponin-containing alfalfa juice is as described in Example 3. Instead of drying, the saponin-free fermentation broth concentrated to 40% solids content - wet feed technology - is preserved with 0.2% formic acid (pH 3.5) until use. The contents of the concentrate, calculated on a dry matter basis, are the same as in Example 3.

Example 5

The saponin-free alfalfa leaf protein concentrate (LFK) prepared in Example 2 is used as a substitute for soy protein as described below.

feeding the Romans. The LEK nycrsfchcrjclartalma: 52.6% crude fiber content: 0% lysine: 4.15% methionine: 0% cisztintartalma: 0%

In broiler feeds (starter and rearing) 15.30 and 45% of recycled soybean was replaced by LFK. During the experiment, the weight gain, the amount of feed used per kg of body weight gain were measured and the health of the animals was monitored.

Housing: 4 storey, cage, random block layout.

Kind of: Hybro, broiler. mixed sex.

Total headcount: 384 headcount, 48 headcount per treatment.

Number of repetitions: 2.

Technology: Normal Hybro Technology, Chickens receive 0-20 days Hybro starter, 21-49 days Hybro breeding abinkkcvcrékcl. Missing mc-59

192,549 thionines were replaced by a synthetic formulation. Λζ housing, feeding, watering, etc. his technique was the same in all groups.

The amount of saponin-free alfalfa protein concentrate (LFK) in the diets fed:

Treatment LFK is the starter LFK in the rearing feed % % control 0 0 Soy protein 15% of LFK 4.1 3.4 Soy protein 30% of LFK 8.2 6.8 Soy protein 45% of LFK 12.3 10.3

The control starter contained 27.0% and the control educator 24.5% soy meal.

These results are shown in Tables I and 2. It can be stated that in the first series of experiments (room I) the best results were obtained in the animals in which 30% of the soybean protein was replaced by saponin-free alfalfa-cone concentrate. In the repeat (Room II), replacing 45% of the soy protein with LFK yielded the best results. The second order is 30%. Based on soybean meal, this represents between 8.2 and 12.3% of the soybean meal in the starter diet and 6.8-10.3% of the soybean meal in the diet.

^ 5 Obesity results in all experimental groups can be considered excellent, no health problems were experienced in any of the treatments.

/. spreadsheet

Feeding LFK with Broiler Chickens 1985. 12.-X. 30

Hall I

Treatment Elhul- MEMORANDUM db Average crowd e Average tömeggyaapodás Average daily weight gain g 1 kg for weight gain forage Live- masses standard deviation s% g % kg % Boot Control _ 452 410 100.0 22.6 1.67 100.0 4.05 8.96 phase Szójafeh. 15% of LFK - 452 410 100.0 22.6 1.67 100.0 5.42 11.98 Szójafeh. 30% of LFK - 460 411 100.2 23.0 1.64 98.2 4.43 9.63 Szójafeh. 45% of LFK - 434 392 95.6 21.7 1.74 106.1 5.15 11.88 Educational Control 3 1796 1344 100.0 48.0 2.49 100.0 19.71 10.97 phase Soybean. 15% of LFK 2 1825 1373 102.2 49.0 2.45 98.4 23.45 12.85 Szójafeh. 30% of LFK - 1846 1385 103.1 49.5 2.39 96.0 18,88 10.23 Szójafeh. 45% of LFK 1 1760 1326 98.7 47.4 2.52 101.2 20.37 11.57

Table 2

Feeding LFK with broiler chickens 1985. 12. —X. 30

II. room

Treatment Being- song db El- wave MEMORANDUM db Average crowd g Consistency weight gain Daily average lömeg- gyarap. g Fodder used per kg bodyweight Live- masses standard deviation s% g % kg % Boot Control 48 1 426 384 100.0 21.3 1.78 100.0 4.09 9.59 phase Soybean. 15% of LFK 48 1 437 395 102.9 21.9 1.73 97.2 4.00 9.15 Soybean whey. 30% of LFK 48 2 465 423 110.1 23.2 1.66 93.2 4.70 10.11 Szójafeh. 45% of LFK 48 - 453 4 1 107.0 22.7 1.67 93.8 4.66 10.28 Educational Control 47 2 1709 1283 100.0 45.8 2.67 100.0 22.40 13.10 phase Soybean. 15% of LFK 47 - 1740 1303 101.6 46.4 2.60 97.4 22.47 12.91 Szójafeh. 30% of LFK 46 2 1785 1320 102.9 47.1 2.64 98.9 21,65 12.13 Szójafeh. 45% of LFK 48 4 1808 13) 0 105.6 48.4 2.49 93.2 20.81 11.51

-611

192,549

Claims (6)

I. Procedure for the partial or total saponin depletion of high saponin-rich alfalfa phylloxera by fermentation, characterized in that Streptomyces sp. Strain GATE S - 38 (accession number MNG 00282) or Penicillium sp. GATE GX-5 strain (letctbchclyczyci number: MNG 00342) or Candida utilis strain GATE G-1 strain (letcibecclycase number: MNG 00341) is preferably used as sole carbon source in the amount of 0.5-0.3% w / v of water-soluble carbohydrate, preparing an inoculum by aerobic, submerged cultivation in an aqueous medium containing an inorganic nitrogen source, an organic biofuel source, and inorganic salts; the resulting inoculum is preferably 5 to 10 micrograms / tcrf. water inoculated with% water-soluble carbohydrate, inorganic nitrogen source, organic biofuel source, inorganic salts and saponin-containing alfalfa press juice, and cultured under acrobic, submerged conditions; the resulting fermentation broth is subjected to heat treatment, filtration and / or concentration, and the saponin-free protein concentrate thus obtained is optionally preserved or powdered. 2. A process according to claim 1, wherein the water-soluble carbon black sugar or starch product or by-product is used. 3. The process according to claim 1, wherein the inorganic nitrogen source is ammonium nitrate and / or ammonium phosphate, and the organic biofuel source organic nitrogen source is cornflour, and / or alfalfa press juice. Process according to claim 1, characterized in that the concentrated saponin-free protein concentrate is preserved by the addition of mineral and / or organic acid, preferably 0.2% formic acid. 5. A process for the preparation of feed for livestock, preferably monogastric stomachs, characterized in that I-4. The supernatant supernatant produced according to any one of claims 1 to 6 is mixed with conventional feed for farm animals. 6. A process for the preparation of feed for livestock, preferably ruminants and monogastric stomachs, characterized in that the method of claim 1 to 4. The press residue obtained during the separation of the saponified protein concentrate produced by the process according to any one of claims 1 to 6, optionally in a preserved state, is mixed with conventional feed for farm animals.