JP3051900B2 - Cellulase preparation for silage preparation and silage preparation method using the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a cellulase preparation for silage preparation and a method for preparing silage using the same. Specifically, cellulase and Trichoderm (Trichoderm) produced by bacteria of the genus Acremonium
a) A cellulase preparation for silage preparation obtained by mixing cellulases produced by bacteria of the genus at a fixed ratio, and an improved silage preparation method using the cellulase preparation for silage preparation.

[0002]

2. Description of the Related Art Various types of cellulases, hemicellulases and amylases have been added as silage enzyme preparations for the purpose of improving the fermentation quality and improving the fermentation quality of silage for the purpose of preparing high quality silage. Are used as single agents or those containing them as a mixture, and further, a mixture of these enzymes and lactic acid bacteria is already on the market for the purpose of enhancing the effect of these enzymes.

According to Japanese Patent Application Laid-Open No. 4-117244, cellulases produced by Trichoderma spp., Cellulases produced by Acremonium spp., And cellulase produced by Aspergillus spp. Cellulases are known.

Japanese Patent Application Laid-Open No. Hei 7-236431 discloses that cellulase produced by bacteria of the genus Acremonium was applied to raw rice straw, for which it is difficult to prepare high-quality silage, and good results were obtained. Have been.

[0005] By the way, the cellulase produced by bacteria of the genus Acremonium described in JP-A-4-117244 is
Compared with cellulase produced by Trichoderma spp. And cellulase produced by Aspergillus spp., The amount of added enzyme (cellulase activity) can improve the quality of distinct silage fermentation with low added amount (low pH, high lactic acid production, Low volatile basic nitrogen production) is known.

On the other hand, as described above, Japanese Unexamined Patent Publication No.
No. 431 discloses that by using a cellulase preparation produced by bacteria of the genus Acremonium, good fermentation quality can be obtained even in raw rice straw where it is difficult to prepare high-quality silage.
It is also known that digestibility is improved when given to animals.

[0007] However, according to the subsequent practical research, it has been found that a cellulase preparation produced by a fungus of the genus Acremonium is produced when a full-scale silage of several hundred tons or more using commonly used grasses of grasses and legumes is prepared. It was found that a considerable amount of juice (drain) was generated.

[0008] This is considered to be due to the fact that the pressure applied to the grass on a real scale is large. However, an enzyme (cellulase preparation produced by bacteria of the genus Acremonium) is added in an amount capable of improving the fermentation quality of silage. In addition, the action of histologically disintegrating the grass is synergistically enhanced, and components that should originally be animal nourishment are lost as waste (drain), and the value of feed is reduced, and fermentation quality and drainage (drain) are reduced. ) It became clear that occurrence was a trade-off.

[0009]

SUMMARY OF THE INVENTION An object of the present invention is to use a cellulase produced by a fungus of the genus Acremonium having a high ability to improve the quality of silage fermentation, and without causing a loss of nutrient components due to drainage. It is an object of the present invention to provide a novel cellulase preparation having an effect of improving fermentation quality of silage and a method for preparing silage using the same.

The present inventors have conducted intensive studies to develop enzyme preparations and silage preparation methods meeting the above-mentioned objectives. As a result, cellulases produced by bacteria belonging to the genus Acremonium and bacteria belonging to the genus Trichoderma were produced. Cellulase in a certain range of ratio, the resulting silage has excellent fermentation quality, but has a small amount of drainage (drain) produced. Was completed.

[0011]

That is, the present invention according to claim 1 provides an avicelase activity ratio between a cellulase produced by a bacterium belonging to the genus Acremonium and a cellulase produced by a bacterium belonging to the genus Trichoderma. And the former
The present invention provides a cellulase preparation for silage preparation, which is blended at a ratio of 0.1 to 9.0 with respect to 1 ).

Next, the present invention according to claim 4 relates to a method wherein the cellulase produced by a bacterium belonging to the genus Acremonium and the cellulase produced by a bacterium belonging to the genus Trichoderma belong to the former 1 in an avicelase activity ratio. In contrast, the latter
An object of the present invention is to provide a method for preparing silage, which comprises treating a silage raw material with a cellulase preparation blended at a ratio of 0.1 to 9.0 .

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.
The cellulase preparation for silage preparation of the present invention according to claim 1, wherein a cellulase produced by a bacterium of the genus Acremonium and a cellulase produced by a bacterium of the genus Trichoderma are blended in a predetermined blending ratio.
It is.

As the bacteria belonging to the genus Acremonium , for example, Acremonium cellulolyticus can be mentioned as a preferred one.

The cellulase produced by the bacteria belonging to the genus Acremonium can be produced, for example, according to the method described in JP-B-60-43945. At this time, after culturing, the supernatant obtained by removing bacteria by centrifugation or the like is further concentrated by ultrafiltration, and water is removed by spray drying to obtain a powder preparation.
At the time of spray drying, lactose, starch, partially decomposed products of starch or glucose are added to the supernatant to improve the process yield and the physical properties of the obtained powder preparation, such as solubility and stability. Improvement is expected.

On the other hand, bacteria belonging to the genus Trichoderma include, for example, Trichoderma viride ( Tricderma viride).
hoderma viride ) and Trichoderma Reese ( Trichoder
ma reesei ), and these can be used alone or in combination of two or more.

[0017] Such a Trichoderma
Cellulase produced by a genus belonging to the genus may be produced according to a general method, for example, the above-described method for producing a cellulase produced by a bacterium belonging to the genus Acremonium.

The cellulase preparation for preparing a silage of the present invention according to the first aspect of the present invention comprises the above-mentioned Acremonium (Acremonium) .
nium) cellulase belonging to the genus of bacteria to produce (hereinafter, often A
Abbreviated as CC. ) And cellulase (hereinafter often abbreviated as TVC) produced by a bacterium of the genus Trichoderma in an avicelase activity ratio with respect to the former 1,
The latter is blended at a ratio of 0.1 to 9.0.

Here, a cellulase produced by a bacterium of the genus Acremonium , ie, ACC, and a cellulase produced by a bacterium of the genus Trichoderma, ie, T
Even if only one of the VCs is used, the object of the present invention cannot be achieved. That is, when only ACC is used, the amount of drainage generated is large,
It reduces the nutritional value of silage. On the other hand, TVC
When only using it, pH becomes high and a high quality silage with excellent fermentation quality cannot be obtained.

The cellulase preparation for silage preparation of the present invention according to the first aspect of the present invention generally has an avicelase activity ratio of 0.1 to 9.0 with respect to the former (ACC) and the latter (TVC) of 0.1 to 9.0.
, Preferably in the avicelase activity ratio, the former (A
CC) 1 and the latter (TVC) in a ratio of 0.2 to 4.0. When the ratio of the former (ACC) is increased to 1: 0.1 or more, the drainage (drain) is significantly increased, while the ratio of the former (ACC) is 1: 1:
If it is below 9.0, the fermentation quality will be reduced,
Neither is preferred.

The avicelase activity, which defines the blending ratio of ACC and TVC, is determined at pH 4.5 (50 mM acetate buffer), using crystalline cellulose as a substrate at a temperature of 50 ° C., and reducing the amount of reducing sugars produced by the DNS method (IUPAC). The activity having an increase in reducing power equivalent to 1 micromol of glucose per minute was defined as 1 unit.

However, the mixing ratio of ACC and TVC is slightly different depending on the kind of the silage raw material to be treated, etc., but the ratio of ACC: TVC = 1: 0.1 to avicelase activity.
9.0, preferably ACC: TVC = 1: 0.2-4.
0.

The cellulase preparation for silage preparation of the present invention according to claim 1 can be a powder preparation as described above as it is as the enzyme dosage form. May be used. Here, as the excipient, for example, flour, soybean flour, starch, partially decomposed product of starch, glucose, lactose, rice bran, bran and the like can be used. The amount of the excipient to be added is usually 10 to 100 times the amount of the cellulase powder formulation, in consideration of the set amount of the cellulase powder formulation to the raw grass and the operability of the addition work to the raw grass. .

The cellulase preparation for silage preparation of the present invention according to the first aspect of the present invention may be any of such powdered preparations, a liquid preparation prepared by dissolving the powdered preparation in a required amount of water, or a powdered preparation at the time of cellulase preparation. Alternatively, a liquid preparation prepared from a concentrated liquid by an ultrafiltration method and a liquid preparation obtained by appropriately diluting the liquid preparation may be used. Liquid products are usually produced by adding sugar alcohols such as sorbitol and reduced maltooligosaccharides as stabilizers to ultrafiltration concentrates, and adding benzoic acid or derivatives thereof or propionic acid as preservatives. You.

The cellulase preparation for silage preparation of the present invention according to claim 1 may be, if necessary, a preparation comprising lactic acid bacteria of the genus Lactobacillus, Streptococcus, Lactococcus, Pediococcus and / or propionibacterium. It can be used in combination with a preparation comprising an acid-producing bacterium of the genus. In this case, there is a further quality improvement effect due to the synergistic action.

Next, a fourth aspect of the present invention is a method for preparing silage, comprising treating a silage raw material with the preparation of the first aspect of the present invention as described above.

Here, the silage raw materials include all of the grasses, feed crops, production by-products, agricultural by-products, feeds and the like used in ordinary silage. First, as grasses, for example, as grasses, timothy, orchard grass, Italian ryegrass, perennial ryegrass, meadow fescue, guinea grass, etc. can be used, and as legume grasses, alfalfa, clover, etc. can be used. . Next, as forage crops, corn, sorghum, barley, rye, triticale, etc. can be used, and as production by-products, molasses, vinegar cake, beer cake, tofu cake, mandarin cake, shochu cake, etc. are used. be able to. Further, wheat, beet top, beet pulp and the like can be used as agricultural by-products. Further, as a feed as a silage raw material, a plain feed, a compound feed, a concentrated feed, a mixed feed obtained by mixing these feeds and a rough feed, which are usually used for fermented feeds, can be used.

When using grass or forage crops as the silage raw material, the raw material grass is cut and used as it is, or after cutting, it is left for 1-2 days and partially dried for use. In this case, the water content in the raw grass is 50-85.
%, Preferably adjusted to 60 to 80%. Thereafter, these raw material grasses can be used after being cut to a length of about several centimeters to a silage method using a cutter or the like. Next, also in the case of production by-products, agricultural by-products, raw material feeds, etc., they can be used after adjusting the water content to the above range.

As a silage method, various methods such as a tower silo, a bunker silo, a stack silo, a roll bale method, and a method using steel or plastic containers can be applied, but a method with high airtightness is preferable.

Incidentally, water can be cited as a factor that greatly affects the fermentation quality of silage. The water content is usually 50 to 85%, preferably 60 to 8%.
Adjust to 0%. Fermentation temperature also has a significant effect on silage quality. From the viewpoint of maintaining fermentation quality and preventing drainage, the storage temperature is 30 ° C. or less, preferably 2 ° C.
It is better to keep the temperature below 5 ° C. In order to suppress deterioration caused by aerobic bacteria during storage, it is fundamental to seal the container and increase anaerobicity. Furthermore, in order to stabilize the quality of silage and to suppress aerobic deterioration after opening, the silage packed with raw grass is usually stored and aged for at least one month, preferably for at least 45 days. desirable.

The present invention according to a fourth aspect is characterized in that the silage raw material as described above is treated using the silage-preparing cellulase preparation according to the first aspect of the present invention as described above. More specifically, the present invention is characterized in that the cellulase preparation for silage preparation according to the present invention as described in claim 1 is added to the above-mentioned silage raw material, and the raw material is treated.

Here, the cellulase preparation for silage preparation of the present invention according to claim 1 is added to the silage raw material in the case of pasture, forage crops, etc., from cutting to cutting.
Raw grass cut to an appropriate length can be packed in a silo and added during the operation up to suppression. In the case of other materials, they can be packed in a silo and added at the time of work up to suppression. As an addition method, a dust may be sprayed and mixed, or a liquid may be sprayed.

The amount of the silage-prepared cellulase preparation of the present invention according to the first aspect of the present invention, ie, the amount of the mixture of ACC and TVC added to the silage raw material is 5 to 300 per kg of fresh raw material based on avicelase activity.
The unit is (U), preferably 10 to 200 units (U). The amount of the mixture of ACC and TVC added to the silage raw material was the avicelase activity and the raw material fresh weight 1
If it is less than 5 U / kg per kg, fermentation tends to be incomplete as a silage, while if it exceeds 300 U / kg, it is not desirable from the viewpoint of increase in drainage and cost performance.

The fermentation quality, that is, the quality of the obtained silage, is preferably such that the pH is low, the lactic acid production is high, the butyric acid production is low, and the ratio of volatile basic nitrogen to total nitrogen is low. It has been. Here, a low pH and an increase in lactic acid production increase the growth of lactic acid bacteria, an increase in butyric acid production increases the growth of butyric acid bacteria, and an increase in the ratio of volatile basic nitrogen to total nitrogen increases the growth of proteolytic bacteria. Each is considered an indicator.

[0035]

Next, the present invention will be described in detail with reference to examples. Example 1 (1) Preparation of Acremonium-derived Cellulase Powder Production culture was performed to obtain an Acremonium-derived cellulase preparation. The medium used was a medium having the following composition, which was heat-sterilized by a conventional method.

[Medium composition] Cellulose 4%, dipotassium hydrogen phosphate 0.16%, bactopeptone 1%, potassium nitrate 0.6%, urea 0.2%, potassium chloride 0.16%,
Magnesium sulfate heptahydrate 0.12%, zinc sulfate heptahydrate 0.001%, manganese sulfate heptahydrate 0.001%,
Copper sulfate pentahydrate 0.001% (pH 4.0).

In 500 mL of the above medium, add acremonium
Cellulolyticus ( Acremonium cellulolyticus TN)
(FERM BP-685) was inoculated, stirred and cultured for 48 hours. Next, the culture solution was used as a seed for 15-liter culture, and the scale-up was further continued. Finally, 300-liter aeration culture was performed for 7 days in a 600-liter tank. After the obtained culture solution was filtered by a filter press, the filtrate was concentrated to 15 liters by an ultrafiltration device, 2 kg of lactose was added, and powdered using a spray dryer. As a result, 5.0 kg of a cellulase powder having an avicelase activity of 851 units / g was obtained.

(2) Preparation of Cellulase Powder Derived from Trichoderma On the other hand, production culture was performed to obtain a cellulase preparation derived from Trichoderma. As the medium, all of the above-mentioned mediums were heat-sterilized by an ordinary method.

[0039] -1) (FERM P-14650 ) was inoculated, stirred for 48 hours, and cultured. Next, the culture was used as a seed for 1
The scale-up was further continued to the 5-liter culture, and finally 300-liter aeration culture was performed for 7 days in a 600-liter tank. After the obtained culture solution was filtered by a filter press, the filtrate was concentrated to 15 liters by an ultrafiltration device, 2 kg of lactose was added, and powdered using a spray drier. As a result, 9.0 kg of a cellulase powder having an avicelase activity of 530 units / g was obtained.

(3) Preparation of mixed powder preparation of Acremonium-derived cellulase powder and Trichoderma-derived cellulase powder Then, Acremonium-derived cellulase powder (AC
C) and cellulase powder derived from Trichoderma sp.
C) and ACC: TVC in an avicelase activity ratio,
They were weighed so as to be 1: 1, 1: 2, 1: 3, and 1: 4, respectively, and sufficiently mixed to prepare a mixed powder preparation.

Various enzyme activity values of the obtained mixed powder preparation (one having ACC and TVC of 1: 1) were as shown in Table 1 below. Various enzyme activities were measured at pH 4.5 (50 mM acetate buffer) and at a temperature of 50 ° C., except for pectinase.
The amount of reducing sugar produced was measured by the DNS method (IUPAC regulation method), and the activity having an increase in reducing power equivalent to 1 micromol of glucose per minute was defined as 1 unit. The pectinase activity was measured at pH 4.5 (5
The activity of reducing the viscosity of pectin (1%) to 75% of the initial viscosity per second was defined as one unit.

[0042]

[Table 1]

Example 2 A predetermined mixing ratio (ACC: TVC = 1: 49 to 1: 0.000) was used in 0.01 M acetate buffer (pH 4.5) to correspond to an addition amount of 50 U per kg of silage raw material. 0
1 ml of a solution in which the ACC-TVC mixture prepared in 5) is dissolved
And 0.2 g of powder (grass dry powder) ground with a 0.1 mm centrifugal grinder after freeze-drying the grass was mixed in a test tube and reacted at 25 ° C. for 2 hours. After stopping the reaction, a 0.01 M acetate buffer (pH 4.5) was added for 2 m.
After adding 1 l and centrifuging (3000 rpm, 10 minutes), the supernatant was diluted 30-fold with distilled water. For this solution, the reducing sugar was measured by the DNS method, and the same treatment was carried out except for the addition of the enzyme and the reaction at 25 ° C. for 2 hours. Table 2 shows the results.

[0044]

[Table 2]

From the results shown in Table 2, ACC: TVC = 1: 0.
In a mixing ratio of 1-1: 9, TVC alone and ACC
It can be seen that the sugar-forming ability is higher than that of the case alone.

Example 3 A solution prepared by dissolving an ACC-TVC (1: 1) mixture in a 0.01 M acetate buffer (pH 4.5) to a predetermined amount (1 to 500 U per kg of silage material). 1 ml
And 0.2 g of dry grass powder prepared in the same manner as in Example 2 were mixed in a test tube and reacted at 25 ° C. for 2 hours. Hereinafter, the amount of reducing sugar produced was measured in the same manner as in Example 2, and the results are shown in Table 3.

[0047]

[Table 3]

From the results shown in Table 3, it can be seen that both of Timothy and Alfalfa produce remarkable reducing sugars when added at 5 U / kg or more.

Example 4 Lactic acid bacteria ( La ) were added to pasture (Timosi first grass) cultivated on a pasture.
ctobacillus casei subsp. r
hamnosus SBT-2300) ( FERM )
^{P-13245), 10 5 cfu} / g · raw grass fresh matter weight) only scatter the experimental group (control group), ACC enzyme preparation solution (53U / kg · raw grass fresh product weight) and the experiments were sprayed with a lactic acid bacteria Ward (ACC Ward), ACC-T described in Example 1
Experimental plots (Experiment plots 1-4) in which a VC mixed powder formulation solution (53 U / kg, raw material fresh weight) and lactic acid bacteria were sprayed, and a TVC enzyme preparation solution (53 U / kg, raw material fresh weight of raw material) and lactic acid bacteria And an experimental plot (TVC plot)
The number n of each treatment section was 2. 200 kg of grass harvested with a harvester cut into 2-3 cm with a cutter
After weighing each, the lactic acid bacteria and the enzyme were mixed uniformly, packed in a 300-liter plastic silo, and sealed with a vinyl film. After storage for one month, the package was opened and the quality of the obtained silage was analyzed. In addition, the dry matter amount of all the obtained seven types of silage was about 20%.

The quality of the silage was evaluated by measuring pH, drainage, organic acid composition (lactic acid, acetic acid, butyric acid, etc.) and the like as follows. That is, after extracting the silage with three times the amount of distilled water, the pH was measured by a glass electrode method, the lactic acid was measured by an enzyme method, and the other organic acids were measured by gas chromatography. For drainage, 10
A fixed amount of sand, gauze, and 10 g of the sample were packed in a 0 ml centrifuge tube in this order, and the weight was determined from the reduced weight of the sample after centrifugation at 500 G for 10 minutes. Table 4 shows the results.

[0051]

Drainage index (%) = [(Experimental zone drainage-Control zone drainage) / (ACC zone drainage-Control zone drainage)] × 100

[0052]

[Table 4]

As shown in Table 4, the pH was controlled by
The value of the ACC section was lower than that of the section C, and all of the ACC-TVC mixed powder preparation sections (Experiment section 1 to Experiment section 4) had ACC.
It was equivalent to the ward. Next, butyric acid, which significantly lowers the fermentation quality, was detected in the control section and the TVC section, but was not detected in the ACC section and the ACC-TV mixed powder preparation section (Experimental Section 1 to Experimental Section 4). Also, judging from the juice index,
It can be seen that the amount of drainage that reduces the nutritional value of the silage is greater in the ACC section than in the control section and the TVC section. However, the amount of effluent produced in the ACC-TVC mixed powder preparations (Experiment 1 to Experiment 4) was lower than that in the ACC.
It can be seen that it decreases at a considerable rate.

To summarize the above trends, ACC-TVC
It can be seen that the mixed powder preparations (Experimental Section 1 to Experimental Section 4) have improved fermentation quality equivalent to the ACC section, and the amount of drainage produced is smaller than in the ACC section.

Example 5 Using the silage prepared in Example 4, a feeding test was conducted with milking cows. That is, 24 milking cows are divided into 4 groups (6 cows in each group), and the TVC section and the silage in the experimental section (the ones in experimental sections 1 to 4) are placed in separate containers before the milking cows in each group. And 1kg was fed at the same time.
The milking cows were allowed to freely ingest for 4 minutes, and the average food intake of each group of 6 cows was measured. Table 5 shows the results.

[0056]

[Table 5]

Example 6 Using the silage prepared in Example 4, a feeding test was conducted with milking cows. That is, 24 milking cows are divided into 4 groups (6 cows in each group), and before each milking cow in each group, the silage in the ACC section and the silage in the experimental section (the ones in experimental sections 1 to 4) are separately stored in separate containers. And 1kg was fed at the same time.
The milking cows were allowed to freely ingest for 4 minutes, and the average food intake of each group of 6 cows was measured. Table 6 shows the results.

[0058]

[Table 6]

From the results shown in Tables 5 and 6, the silages obtained with the cellulase preparation for silage preparation of the present invention (Experimental sections 1 to 4) were the same as those obtained by the conventional treatment with TVC alone (TVC section). It can be seen that the amount of food consumed is clearly higher than that of the silage (ACC section) obtained by the ACC treatment alone. Therefore, in consideration of the results in Table 4, etc., according to the cellulase preparation for silage preparation of the present invention, not only the fermentation quality is high, and there is no loss of nutrient components due to drainage, but also the taste of cattle is improved. It is considered that a suitable silage is obtained, and it is recognized that the feed value is high.

Example 7 An experimental plot (control plot) in which only the lactic acid bacterium described in Example 4 (10 ⁵ cfu / g, raw material fresh weight) was sprayed on pasture cultivated on a pasture (Timothy No. 1 grass), and a pasture. ACC-TVC mixed powder formulation solution described in Example 1 (mixing ratio 1: 1, avirase activity 1)
3.3, 26.5, 53.0, 106.0 U / kg, fresh material weight) and lactic acid bacteria were sprayed on the experimental plots (experimental plots A to A).
D) and an experimental plot (TVC plot) in which the TVC enzyme preparation solution (0.01%) described in Example 4 and lactic acid bacteria were sprayed on the grass (Timothy No. 1 grass) cultivated on the ranch. Was set to 2. Grass harvested with a harvester was cut into 2-3 cm pieces with a cutter, weighed 200 kg at a time, uniformly mixed with lactic acid bacteria and enzymes, packed in a 300-liter plastic silo, and sealed with a vinyl film. After storage for one month, the package was opened and the quality of the obtained silage was analyzed. All of the six silages obtained had a dry matter of about 20%. Table 7 shows the results.

[0061]

[Table 7]

As shown in Table 7, the pH of the ACC-TVC mixed powder preparations (Experiment A to Experiment D) was lower than that of the control. In the ACC-TVC mixed powder preparations (Experiment A to Experiment D), as the amount of application increased,
pH was lower and lactic acid content increased. ACC
-In the TVC mixed powder preparations (Experiment A to Experiment D), a small amount of enzyme was added (1/4 of the activity in the TVC),
Silage quality equivalent to that of the VC section was obtained.

[0063]

According to the present invention, the cellulase preparation for silage preparation of the present invention is a mixture of a cellulase produced by a bacterium belonging to the genus Acremonium and a cellulase produced by a bacterium belonging to the genus Trichoderma in a certain ratio. The fermentation quality of the obtained silage is excellent, but the amount of drainage generated is small.

That is, the cellulase preparation for silage preparation of the present invention according to claim 1 uses a cellulase produced by a bacterium belonging to the genus Acremonium having a high ability to improve the quality of silage fermentation, and without causing loss of nutrient components due to drainage. , Has the effect of improving the fermentation quality of silage.

Further, the cellulase preparation for silage preparation of the present invention according to claim 1 can improve fermentation quality of silage with a small amount of enzyme (cellulase activity) added.

Further, the method for preparing silage of the present invention according to claim 4 has a high ability to improve the quality of fermented silage, and furthermore, there is little loss of nutrients due to drainage. The silage thus obtained has a high fermentation quality, a small loss of nutrients due to drainage, and a high feed value.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI C12R 1: 885) (72) Inventor Toru Hamaya 5-3-1 Chiyoda, Sakado City, Saitama Meiji Seika Co., Ltd. 72) Inventor Toshiaki Kono 5-3-1 Chiyoda, Sakado City, Saitama Prefecture Meiji Seika Co., Ltd.Biological Science Research Institute (72) Inventor Hidetoshi Kubota 580 Horikawa-cho, Saiwai-ku, Kawasaki City, Kanagawa Prefecture Meiji Seika Co., Ltd. (72) Inventor Shunji Miura 36-1, Nishinohoro, Ebetsu-shi, Hokkaido Snow Brand Seedlings, Inc.Technical Research Institute Co., Ltd. (72) Inventor Toru Kitamura 36-1, Nishinohoro, Ebetsu-shi, Hokkaido, Japan Snow Technical Seedling Co., Ltd. 36-1 Nishinohoro, Yokohama City Snow Seed Seed Examiner at Technical Research Institute, Inc. Mayumi Saito (56) References JP-A-4-117244 (JP, A) (58)査the field (Int.Cl. ^7, DB name) C12N 9/00 - 9/99 A23K 3/03 BIOSIS (DIALOG) WPI (DIALOG)

Claims (4)

(57) [Claims] An avicelase activity ratio between cellulase produced by a bacterium of the genus Acremonium and cellulase produced by a bacterium of the genus Trichoderma.
In the former 1, the latter is blended in a ratio of 0.1 to 9.0.
A cellulase preparation for preparing a silage. Wherein acremonium (Acremonium) genus bacteria, Acremonium cellulolyticus (Acremonium c
silage for cellulase preparation according to claim 1 wherein the ellulolyticus). 3. The bacterium of the genus Trichoderma comprises Trichoderma viride and / or Trichoderma reesei .
The cellulase preparation for silage preparation according to claim 1 or 2, which is: 4. An avicelase activity ratio of a cellulase produced by a bacterium of the genus Acremonium to a cellulase produced by a bacterium of the genus Trichoderma.
In the former 1, the latter is blended in a ratio of 0.1 to 9.0.
A method for preparing silage, comprising treating a silage raw material with the cellulase preparation prepared as described above.