NL8000876A - METHOD AND PRODUCT FOR THE CONSERVATION OF GREEN PLANTS AND THE MOISTURE-CONTAINING BY-PRODUCTS OF THE FEEDING INDUSTRY - Google Patents

Description

<t 80 3049 / BZ / asm

Applicant jSociété CEVA S.A., 14 rue Garnier - 92200 NEUILLY-SUR-SEINE (France).

Short designation jProcess and product for the preservation of green plants as well as the moisture-containing by-products of the animal feed industry.

The invention relates to a method for preserving and making fresh plants valuable, in order to make it possible to use them in tasty form after harvesting, depending on the need, the invention furthermore relates to a composition for carrying out these method.

5 The importance of preserving green plants has led to a large number of studies devoted to the associated difficulties.

Storage in silos is based on the preservation of fresh plants for a short or long period in a closed, air-tight space and in an acidic environment to prevent the development of alkali and gaseous decay bacteria that cannot develop in an acidic milJLeu. to go. Good silo preservation is difficult to implement and faces many problems, the most important of which are the formation of fermentation products which are dangerous to the health of animals and thus of the people who use the meat and the milk products.

The intended working method is as follows. By having lactic acid bacteria produce lactic acid from the soluble sugars present, the pH of the environment decreases to about 4. However, it appears that in most cases the enzilized plants contain insufficiently fermentable sugars, so that the development of the lactic acid bacteria present in the environment is insufficient. , the pH does not drop quickly enough to 4, and butyric acid bacteria and anaerobic bacteria develop, which convert the sugar residues into butyric acid, acetic acid, carbon dioxide and hydrogen, decomposing the proteins into ammonia 800 0 8 76 - 2 - and other metabolic products turn into.

A number of solutions have been proposed for this problem, in particular chemical acidification by adding acids. However, this method is not without dangers. Organic acidification is also carried out by adding bacteria that deplete carbohydrates; but this biological acidification is very difficult to carry out.

Furthermore, methods have been proposed in which a raw material rich in carbohydrates, for example molasses, is added to the silo content, but the results obtained are irregular and the process is expensive to use.

French Pat. No. 2,361,828 describes a method for preserving and distributing green plants by acidification with lactic acid, adding to the plants bacteria capable of initiating lactic acid fermentation, prior to ensilation. additive which converts the higher carbohydrates into fermentable sugars used by the bacteria causing fermentation of lactic acid. This degradation factor consists of Grann bacteria, which do not ferment starch, glucose, mannitol, rhamnose, sucrose, amygialine, arabinose, but maltose, inositol and sorbitol. These bacteria are VP +, oxidase +, catalase +, nitrates +, urea, citrates, indole and I ^ S-. The degradation factor may also contain one or more enzymes capable of splitting polysaccharides, especially starches, pentosans and other polysaccharide complexes, into fermentable sugars. In particular, according to this patent, a fungal hemicellulolytic complex can be added to the feed, which has essentially a galactomannan-degrading action in addition to some other secondary actions such as, for example, a xylanase, amylase, pectinase and an amylase which is used to form provides the maltose necessary for the preparation of lactic acid by bacteria.

French Patent 2,930,908 is an improvement on Patent 2,361,828. While the amylase used in French Patent 2,361,828 is a fungal exoamylase that does not completely decompose starch, the amylase of Patent 2,390,908 is a bacterial amylase. This enzyme (endoamylase) is active against liquid starches and mainly supplies c ^ D-maltose and a small amount of c ^ -D-glucose, at a pH of 5 to 8. French patent 2,390,908 also describes the the use of amyl glucosidase, which is much more effective compared to the amylose and amylopectin chains in the preparation of CF-D-glucose. The enzyme composition of this patent also contains a fungal hemicellulolytic complex which reacts 70 with the polysaccharide constituents of the cell membrane and with dextrins at pH 5.5 to 2.

The present invention aims to provide a method as well as an improved composition for storing plants in silos, the improvement relating to both the enzymes 75 and the bacterial composition. The enzyme composition is distinguished by the cellulolytic complex or cellulose added to the previously used enzymes, which is able to cleave the (B) bond (H3) and (1- * 4) of polysaccharides which are not affected by the said enzyme composition. .

It is a fungal cellulolytic complex that decomposes natural cellulose and other polysaccharides present in cell membranes.

This enzyme is active at a pH of 2 to 5. Between the cellulolytic complex and the hemicellulolytic complex there is a synergistic action aimed at the decomposition of vegetable structures.

Due to the location of the cellulose in the plant, the hydrolysis S has to be carried out with different enzymes or with one enzyme that performs different functions, namely: 30 “hydrolysis of the amorphous zones - swelling of the crystalline areas and breaking of the hydrogen bonds to form linear molecules, 800 0 8 76 - 4 - - hydrolysis of cellobiose to glucose - transglucosylation of the di- and oligosaccharides and breaking of the irregular bonds in cellulose and of the primary hydrolysis products.

The cellulolytic complex has been selected depending on the above conditions and after a large number of experiments with different plants.

A cellulose is defined by its activity expressed in international units per gram of product, where an international unit (I.E.) is the amount of enzyme which liberates per minute the sugar equivalent of a micromallucose reducing / of a given cellulose substrate.

The efficacy depends on the substrate used: - the efficacy is obtained in tests with Whatman CC 31 paper powder; - and the efficacy C1 in tests with carboxymethyl cellulose.

The complexes used according to the invention have a very strong action with respect to cellulose and a secondary action with respect to hemicellulose. The high cellulolytic capacity is evidenced by 2 (), the very strong actions C C and C ^ and by the cellobiase activity (SS-glucosidase).

The activity of the enzyme complex Cj, according to a hypothesis composed of the activity Cj, plus an unknown factor of action is evidenced in particular by the solubility to cellulose.

C1 is capable of decomposing pure cellulose (crystalline cellulose with a degree of polymerization P.G> 3,500 glucose units) into amorphous cellulose.

More specifically, the action of the enzyme complex C follows

X

3 expresses the reducing capacity with respect to endo and exoglucanases.

These enzymes act primarily on cellulose fibers in zones of low crystallinity, breaking the long glucose chains into much smaller units.

800 0 8 76 ♦ i - 5 -

The activity of β-glucosidase results from the action on cellobiose units generated by the action of C ^.

These different activities cannot be taken separately. For the decomposition of these homopolymers, there is a synergistic effect between and C 1. The enzymatic action of C leads to the formation of cellobiose, which is then hydrolyzed by 8-glucosidase x to form two glucose units.

The hydrolysis of cellulose appears to proceed according to the following schedule: r 10 Native Cellulose ..... - "-} reactive cellulose C2 hydrolysis

Reactive cellulose ............) cellobiose beta-glucosidase. . 1

Cellobiose Nyaroalysis ^ 2 molecules of glucose

The C 1 complex dissolves the crystalline cellulose (polymerization-15 degree (P.G.) over 3500 glucose monomers) to yield reactive cellulose (amorphous cellulose).

Due to the action of the enzyme complex C (endo and exoglucanases)

X

the reactive cellulose chains are broken, i.e. internally and at endoglucanases at random sites, or in the case of exoglucanases (first) at the non-reducing ends.

These reactions yield cellobiose units.

The cellobiose units are hydrolyzed by β-glucosidase to form two glucose molecules.

In the fungal cellulase according to the invention, the enzyme complexes C1 and C1 are capable of hydrolyzing the insoluble pure cellulose to glucose.

The cellulases according to the invention are selected not only depending on the general conditions indicated above, but also taking into account the local silo environment, depending on the polysaccharolytic behavior of these cellulases.

The polysaccharolytic behavior is determined on two βηη n fi 76 - 6 - substrate types: a) - pure substrates b) - anhydrous substrates.

The pure substrates: The activity of the cellulases A, B, C, D, E, F on different substrates is determined at certain concentrations in a 10 µM acetate buffer medium.

The activity is determined under known conditions, ie at pH 4, 8 and 50 ° C, after a hydrolysis lasting 20 minutes. The reducing power is determined using the hexaferrocyanide method.

-1-ï-r — i - ^ - 1 — f paper C.M.C. Pectin Xylane Arabino Starch Locus Wathman galactan bean substrate concentrates- 1 ^ 1 ^ 0.25 ^ 0.25 ^ 0.10 ^ 0f25% Of25% tie _____; ___ enzymes ΙΕ / g ΙΕ / g ΙΕ / g ΙΕ / g ΙΕ / g ΙΕ / g lE / g (efficacy (C.) (C) ^ ness) __ 1 X________ A *> - 400 6444 356 511 22 394 544 B 211 5660 322 611 33 100 300 ~ C 266 1999 411 311 0 244 389 “5 52 6440 255 511 0 120 132 'r“ Ê 372 8500 305 605 0 0 0 F 960 7666 1030 1200 Ö Ö ~ 140.

80 0 0 8 76

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Anhydrous substrates.

The activity of the enzymes A, B, C, D, E and F is also determined on various dehydrated vegetable substrates.

The activity was tested at pH 3, 4.8-5.8 and o at 30 ° C after 24 hours hydrolysis. The reducing power is determined using the dinitro-salicylic acid method.

r —— I ----- examined Lucerne Ray-grass Tender Sugar Beet Pulp Straw "'Sjjbstraten examined"'. Cellulose pH Efficacy (lE / g) - ______ ff _ A 3.8 2810 2280 1560 1810 1560 4.8 2440 2560 2080 1530 1690 __5.8 1560 2220 1690 1500 1530 B 3.8 1560 1670 1530 1220 1390 4.8 1280 2060 1670 1220 1500 5.8 890 1680 1940 890 1280 C 3.8 1280 1530 1390 ÜÏÖ ÏÏÏÖÖ 4.8 1280 1750 1530 1110 1290 __5.8 940 1560 1170 720 1110 D 3.8 1390 1440 1470 1278 U67 4.8 1190 2060 1390 1111 1306 __5.8 890 1720__1530 972 1222 E 3.8 611 1190 1111 556 694 "4.8 972 1528 1250 0 694 5.8 0 1167 1278 0 694 F Ti 1306 1306 500 694 556 4.8 889 972 1222 610 444 5.8 750 1000__560 550 333 800 0 8 76 - 8 -

From the analysis of the different results one can determine the effectiveness of the cellulose complex intended for storage in silos according to the invention.

It has thus been found that cellulose or the cellulose complex added according to the method of the invention to the fresh plants intended for storage in silos must have a C 1 activity of 50-0.005 I.E. and a C activity of x0 500 to 0.05 I.E. per 100 g of plant, at a pH of 4.8 and 50 C after a 20-minute hydrolysis.

In addition, even after a long stay in the silo, the selected cellulose must retain considerable efficacy.

It has been experimentally shown that under conditions such as that which occur approximately in silos, the cellulose complex must still have at least 30% of its original activity after 10 days.

Preservation time in days 0 5 10 15 20 pH 4.8 100 * 97 * 61 * 26% 3 * pH 5.8 100 * 85 * 58 * 27 * 0 *

The rate of enzymatic degradation of the plant structure 20 depends on the removal of glucose, this displacement of the enzymatic equilibrium is achieved with bacteria with a high fermentation capacity, whereby the environment is acidified very quickly. Once the vegetable mass has stabilized, the growth of the bacteria and the formation of lactic acid are stopped, in contrast to the cellulolytic action of cellulose, which continues for more than 10 days.

This action will increase the nutritional value of the enzilized plant product by the formation and accumulation of glucose in the silo product.

Thus, the enzyme composition used according to the method of the present invention comprises a cellulose or a cellulolytic complex having the above characteristics, together with the enzymes described in French Pat. No. 2,390,908, ie a fungal amylase, a bacterial amylase, an amylogiucosidase and a hemicellololytic complex.

These enzymes present in the composition thus have an additional effect due to their action on the very complex to very simple sugars at pH values of 7 to 2. The maximum activity of each enzyme depends on the reduction of the pH value. , those in the silo not below pH 3.5 and at temperatures of 10 to 30 ° C associated with the conditions in the silo.

The bacterial composition according to the invention is distinguished by the use of new strains of bacteria that are used in plant ensilaging, wherein these strains can be used alone or in admixture with known and strains of bacteria and / or enzymes mentioned here. This new formulation results in much improved results when en-silencing. The new strains of bacteria used according to the present invention are gram bacteria belonging to the Enterobacteriaceae family. the group Erwinia or Pectobacterium to the group Herbicola, which are referred to as Enterobacter agglomerans (classification Bergey's 20 Manual of Bacteriology, 8th edition).

The properties of these bacteria are shown in the following table.

Table A

Enterobacter Agglomerans

Gram - Arabinose + 25 Malonate use + Maltose

Glucose + Trehalose +

Phenylalamine Desaminase - Xylose 0NPG + Starch +

Indole - Oxidase 30 - Gelatin + LDC - Amygdalin + 0DC - Melibiosis 800 0 8 76 - 10 -

Urease - V.P. +

Saccharose + Rhamnose

Arginine Dihydrolase -Salicin 5 Adonitol Isositol Sorbitol

The use of these new bacterial strains for plant ensilaging, alone or together with other bacterial strains and / or enzymes, is carried out according to the known methods described in the literature. The bacteria can be grown on a nutrient medium consisting of grain meal types. The composition to be placed in the silo comprises a mixture of the necessary bacteria on a soluble or insoluble carrier with the mixture of enzymes prescribed according to the present invention.

According to a preferred embodiment of the invention, the enzymes and bacteria are applied to a preferably very finely ground grain (wheat, barley, sprouted barley) carrier.

In another preferred embodiment, the bacteria and enzymes are applied to a support consisting of pregelatinized starch, maltohexose, etc., which are readily soluble in cold water.

Starch and other polysaccharides present in the carriers increase the carbohydrate content in the plants and thus increase the nutritional value of the enzilized product. The composition containing the mixture of the bacteria on a carrier and the enzymatic complex, whether or not on a carrier of grains, may contain these constituents in different amounts, the dry mixture starting per gram in an amount of 100,000 to 1,000,000 cocci and 100,000 to 1,000,000 bacilli.

The amounts of cellulose, hemicellulolytic complex, amylase and amyloglucosidase can run out and are calculated 800 0 8 76 r - 11 depending on the nature of the feed to be treated.

The amount of cellulose added to the fresh plants to be ensilated must correspond to an activity of 50 to 00000 I.U. per 100 g of vegetable material (i.e., 4-5 to 2 wt. 2 wt. of fresh plant at a cellulose with a content of 250 IU / g. The amounts of the other enzymes are between 0.05 and 0.02. plant material) bacterial amylases at 250 units per gram; between 0.1 and 0.2 wt. vegetable matter in fungal amylase with 50,000 units of PS 50 / g; between 0.1 and 0.7 wt% of vegetable matter in amyloglucosidase at 200 units AG / g and 0.05 to 0.2 wt% of the vegetable material for the hemicellulose complex at 35,000 IU / g.

When the enzymes according to a preferred embodiment of the present invention are applied to a grain carrier, the product to be introduced into the material to be ensilated can contain 1 kg of enzymes per about 9 kg of carrier. With a soluble carrier, the ratio of the amounts of enzyme to carrier is 1 / 1.5 to 1/4.

The incorporation of cellulose into the carrier increases the amount of fermentable sugar available, resulting in more and faster lactic acid. The vegetable protein will then be better preserved. Furthermore, the nutritional value of the medium is increased by this.

To illustrate the importance of the cellulolytic complex and of the bacteria of the Enterobacteriaceae family, experiments have been carried out in micro-silos fin in mini silos. The different preservation parameters have been investigated as well as those relating to the vegetable structure.

1. Tests in microsilos.

A first series of tests was performed in micro-silos in the laboratory.

These tests were carried out in the month of May with lucerne; in micro-silos with a capacity of 1 kg. The lucerne is finely chopped and then anilized after 76 - 12 - using the method described below: A-Men uses a number of microsilos using a preservative additive with the following algae composition: grain carrier 90 $ 5 premix 10 $

The premix consists of: - a bacteria complex (five lactic acid bacteria on a lactose carrier); - an enzyme complex consisting of: a fungal amylase, a bacterial amyulase, an amyloglucosidase, a fungal hemicellulolytic complex, the main property of which is galactonammanase activity; 15 an energy-rich carrier; finely ground barley + milk serum; and the bacteria and enzyme complexes each of which are included in the premix in an amount of $ 1.

The preservative additive was added to the lucerne 20 in an amount of $ 2 (in an amount in the premix of thus 0.2 $), and 1 $ of barley was added.

B-Men use a number of microsilos with the same preservative additive as under A, however adding a fungal cellulose (Trichodemaviride) with an activity of 250 units / g and an activity of 2500 units / g (determined at a pH of 25 4.8 and 50 ° C at a reaction time of 20 min.) Adding the cellulose in an amount of 2 g / T (or 2.10 wt.%) Calculated on the fresh plants (test B) and 200 g / T (or 0.02 $) based on the fresh plants (test B ') C-Men use a number of blank microsilos, which are filled with 30 lucerne without preservative: T = 0.

The microsilos are opened after 100 days, with the following parameters: pH, lactic acid, NH ^ / N total.

The results are summarized in the following table: 80 0 0 8 76 - 13 - pH lactic acid in ΝΗ ,, / Ν total

___ g / kg MS__J

Blank: T = 0 5.66 26 g 25.15

Lucerne + 0.2 # of 4.13 143.68 16.-33

premix A

Lucerne + 0 * 2 # of the premix with 4.32 144.75. 16.20 fungal cellulase 2 g / T (B) ______

Lucerne + 0.2 # of the premix with 3.73 169 47 15.02 fungal cellulase 200 g / T (B ') __

It can be seen that adding a cellulose to the bacterial and enzyme complex of patent 2,390,908 leads to an improvement of the preservative activity. A better preservation of the plants is obtained, which is shown by comparing A with B * from: - a decrease in pH: 4.13 to 3.73 (10 # less) - an increase in lactic acid production of 15 # - a reduction of 8 # in the ratio of NHg / N total; resulting in a better preservation of the vegetable protein, resulting in faster acidification of the environment.

2 - Tasting with mini silos.

These tests were carried out in June with lucerne in mini silos with a volume of 150 kg. The lucerne is chopped and then placed in a silo according to the method described below.

A - A number of mini silos are used using the same preservative as the micro silos under A and the amount of premix added is 0.2 #.

B - A number of mini silos of the composition and amount indicated under A are used, to which the same fungal cellulase in 800 0 8 76-14 is added in an amount of 2 g / T.

After 100 days of ensilation, the mini silos are opened, with analysis of the content yielding the following results.

v "** -—...............-

A B

silos with 0.2 $ silos with 0.2 $ premix mixture + cellulose --- iMÜ_

Dry matter (MS) $ 23.67 27.27 E.N.A. (gAg MS) 525257979_5794

Lactic acid g / kg MS 80.62 114.40

Acetic acid gAg MS 17.42 20.04 NH3 / N total 18.40 13.31 N total gAg MS ~ 21.70 23.12 N ammonia gAg MS 3.87 3.09

The above results clearly demonstrate the importance of adding a cellulose to the bacteria + enzyme complex described in U.S. Pat. No. 2,390,908.

The addition causes: - a lower pH of 3.94 compared to 4.2 ($ 6 less) - a considerably larger amount of lactic acid ($ 29.53 more) - the protein part of the enzilized product is much better preserved if one still uses a cellulose which confirms the analysis. In this case one has: - a smaller ratio of NH 4 / N total ($ 27.6 less) - a higher protein content ($ 6 more) - a lower ammonia nitrogen content ($ 20.1 less).

This much faster acidification rate can be explained by the fact that the bacteria have a considerable amount of fermentable sugars, which can easily be converted into lactic acid, more quickly. This availability shows the effect of the cellulose added to the environment. In the following examples, the supported mixture according to the method of the present invention consists of the complex described in French Patent 2,390,908 and / or Patent 2,361,828; the enzymatic complex described in the present invention, to which gram braces of the Enterobacteriaceae family of the genus Erwinia or Pectobacterium, from the group Herbicola, are added, with 10 to 10 bacteria / g being added to the mixture.

Examples.

3

The tests are carried out in silos with a capacity of 4 m with Lucifer fingerwort harvested in June at the beginning of the earth formation. The feed is chopped and stored under four different conditions: A: ensilage without treatment B: in a silo with 4 liters per ton of formic acid according to a known method.

C: in a silo with the premix, the composition of which is described in French Pat. Nos. 2,631,828 and 2,390,908: wherein 7 kg of a culture of 7 bacteria is added on a support, the properties of which are indicated in French Patent 2,361 .828, p. 7, No. 1 to 7, and 10 kg of enzymes per ton.

D: in a silo with the premix according to C, to which the bacteria according to the invention are added: gram brackets of the family of Enterobacteriaceae of the genus Erwinia or Pectobacterium, from the group, Herbicola, designated Enterobacter agglomerans. The amount of complex added is 10 kg / T.

After 90 to 100 days, the different silos are opened and the vegetable products are analyzed. The results are summarized in Table B.

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It can be deduced from the results that the mixture according to French patents 2,361,828 and 2,390,908 already shows an improvement in the preservation of the silo content, which is evident from: - a decrease in the pH from 5.28 to 4 , 02;

5 - an increase in the lactic acid content from 23 to 90 t g / kg MS

- a better protection of the proteins, which is evidenced by a decrease in the N-MIg ratio from 17.76 to 14; and the total N position soluble from 63.8 to 48.1.

N total 10 The mixture has the advantage that no propionic acid and butyric acid are formed.

The mixture according to the invention provides a greater improvement, which is evident from: - a decrease in the pH from 4.02 to 3.7

15 - an increase in the lactic acid content from 90 g to 112 g / kg MS

- better protection of the proteins, which is evidenced by a reduction in the ratio N-NH 2 / N total from 14 to 8; and the ratio of N soluble / N total from 48.10 to 44.

The mixture has the advantage that no propionic acid and butyric acid are formed.

The digestibility and uptake of silo production has been investigated in sheep. After several treatments, it was found: B: when treated with formic acid: 0.64 UF / kg C: when treated according to French patents 2,361,828 and 2,390,908: 0.74 UF / kg D: when treated according to the invention: 0.82 UF / kg

The nitrogen balance in growing animals leads to results from which the importance of the treatment with the products from the silo according to the present invention follows, as can be seen in table 30 C.

800 0 8 76 -18-

__ TABLE C

Ml i '~ I' 1 ....... m j * ------------ - - parameters N ... - .... ..

. N in faeces N in urine N absorbed mgeno · -....... * - i -? —-

r treat! ng ^ - ^^ g / y input / j input / j JÖM

g / y nomen nomen taken £ _______ nomen? A: no treatment 21 7.90 3.76 13 61.9 0.10 0.50 B: with 0.4% ants - 18.9 7.34 38.8 10.4 55 1.1 5.88 acid _______ C: according to main 17 7.07 41.6 8.12 47.7 1.8 10.5 patent and 1st supplement D: according to present 16.5 5.4 38.8 7.70 47 2.40 14.5 current invention

The nitrogen retained in the present invention is more than twice as high as in the treatment with formic acid.

80 0 0 8 76

Claims (10)

1. A method for preserving and making green plants valuable by acidifying them with lactic acid, whereby a factor is added to the plants for ensilering in a silo for breaking down higher carbohydrates into fermentable carbohydrates which can be used by the bacteria used for the lactic acid fermentation, which factor consists of an enzyme complex and / or a bacteria complex, characterized in that the enzyme complex may or may not be mixed with a fungal amylase, a bacterial amylase, an amyloglucosidase or a fungal hemicellulolytic complex, the main property of which is a galactomannanase activity. Containing a cellulose or a fungal cellulolytic complex, it is capable of converting native cellulose to glucose with an activity of 50-0.005 international units and a C activity of 500 to 0.05 international units per 100 g of fresh plants. under 15 years of prevailing conditions in ensilering at least 30 ^ of them after 10 days has retained activity, the bacteria complex containing gram bacteria, which do not ferment starch but maltose, belonging to the family of Enterobacteriazeae, strain Erwinia or Pectobacterium, group Herbicola, designated 20 Enterobacter agglomerans. 2. Process according to claim 1, characterized in that the bacteria of the family Enterobacteriaceae are used simultaneously with bacteria known for this purpose which are capable of converting the higher carbohydrates into fermentable carbohydrates · Method according to claim 1 and / or 2, characterized in that the bacteria of the family Enterobacteriaceae are used simultaneously with gram + bacteria containing starch, glucose, mannitol, rahmnose, sucrose, amygdalin and arabinose, but on our own maltose, inositol and sorbitol do not ferment * 4. Process according to claim 1, characterized in that the enzyme mixture consists of: 2.10 "to 2% cellulose with 250 international units activity per g; 0.05 to 0.2% o bacteria amylase at 250 units per g; 0.10-0.20 ^ fungal amylase of 5000 units PS 50 / g; 0.10 to 0.70 µS amyloglucosidase with 200 units A6 / g; 0.05 to 0.2% o hemicellulose complex with 35000 national units / g; where% o refers to the weight amount of the 10 enzilized plants * 5. A method according to claims 1 and / or 2, characterized in that the enzymes are applied to a grain carrier in an amount of about 1 kg of enzymes per 9 kg of carrier. 6 * Method according to claim 1, characterized in that the bacteria are grown on a carrier consisting of cereal flour * A method according to claims 1 and / or 2, characterized in that the bacteria and the enzymes are applied on a carrier consisting of grain starch, gelatinized starch or malto-hexose, soluble in cold water, in an amount of 1 kg of bacteria and enzymes per 1.5 kg to 4 kg carrier · ' Method according to one or more of claims 1 to 5, characterized in that the enzymes and bacteria culture on their support are added in an amount of 10 to 15 kg / tonne of plants to be ensilated in a dry state, said additive 10 ^ contains up to 10 ^ live germs per g. 9 * Composition for preserving and making valuable 8 0 0 0 8 76 - 21 - green plants, containing lactic acid-producing bacterial strains and a factor for breaking down higher carbohydrates into fermentable carbohydrates, consisting of an enzyme complex and / or a bacteria complex, characterized in that the enzyme complex, whether or not scanned with a fungal amylase, a bacterial amylase, an amyloglucosidase or a fungal hemicellulolytic complex, the main property of which is a reactant lactomannanase active, a cellulose or fungal cellulolytic complex which is capable of convert cellulose into glucose with a 10 Ci activity of 50 to 0.005 international units and a C • activity of 500 to 0.05 international units per 100 gx, under the fresh plants and / or under the ensilage conditions, has retained at least 30 µ of its activity after 10 days, wherein the bacteria complex contains gram bacteria, which do not ferment starch as much as 15 maltose, belonging to the family of the Enterobacter teriaceae, genus Erwinia or Pectobacterium, group Herbicola, designated Enterobacteriobacter agglomerans. 9 -y - 19 CONCLUSIONS 10. A composition according to claim 9, characterized in that the gram-enterobacteriaceae bacteria are united with bacteria capable of converting higher carbohydrates into fermentable sugars and in particular gram-t bacteria containing starch, glucose, mannitol, rhamnose, sucrose ^ amygdalin and arabinose yes ^ but do not convert maltose, inositol and sorbitol · on η nr 76