A composite enzyme preparation for efficiently degrading fiber and a preparation method and application thereof

Abstract

The application relates to the technical field of microbial fermentation, and particularly discloses a composite enzyme preparation for efficiently degrading fibers as well as a preparation method and application thereof. The composite enzyme preparation is obtained by solid-state fermentation of Aspergillus oryzae, and the composite enzyme preparation comprises xylanase, beta-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease. Further, the solid culture medium used for the solid-state fermentation comprises the following components in parts by weight: bran 5-10 parts, soybean meal 2-4 parts and corn flour 0.2-1 part. The application further provides a preparation method of the composite enzyme preparation, which comprises preparing a spore suspension of Aspergillus oryzae and solid-state fermentation culture. The composite enzyme preparation has a significant degrading effect on fibers. When the composite enzyme preparation is used in animal feed, the digestibility of fiber feed by animals can be improved, the growth performance and health condition of the animals can be improved, the breeding cost can be reduced, and the economic benefits of a breeding farm can be improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microbial fermentation, in particular to a compound enzyme preparation for efficiently degrading fiber and a preparation method and application thereof. BACKGROUND

[0002] With the rapid development of animal husbandry, the problem of feed shortage in China is becoming increasingly serious. Since high-energy value grains are mainly used for human consumption, more high-fiber grain processing by-products, such as corn stalks, are used in animal feed. These agricultural by-products are rich in organic matter and are considered important renewable resources. However, the fiber contained in these by-products poses a certain challenge to animal digestion, and therefore the economic value of using agricultural by-products as animal feed cannot be fully realized.

[0003] At present, in order to improve the utilization rate of high-fiber feed, the industry mainly adopts physical, chemical and biological methods to treat fiber feed. Among them, the physical method changes the fiber structure through heat treatment, high-pressure cooking and other methods to improve the enzymatic efficiency of fiber, so that it is easily absorbed by animals. However, this method has very limited effect on fiber and requires a large amount of energy consumption, so it cannot be applied on a large scale. The chemical method uses acid or alkali solution to break the chemical bonds between fiber molecules to improve the degradation efficiency of fiber, but the use of chemical reagents increases the processing cost and causes environmental pollution. The biological method uses enzymatic action of enzyme preparation to decompose complex fiber components into small molecules that are more easily absorbed by animals, and has low operating cost and little environmental pollution, so it is the most widely used method for treating fiber feed.

[0004] However, although the biological method reduces the processing cost of fiber feed to some extent, it still has the following problems: first, the effect of a single enzyme in the enzymatic process is limited, and it is difficult to effectively deal with the complex structure of fiber, resulting in low enzymatic efficiency; second, the existing commercial enzyme preparation is expensive, which limits its large-scale application. Therefore, if enzymes can be produced and used on site, it is of great significance to develop a high-efficiency, low-cost compound enzyme preparation for improving the economy of fiber feed. SUMMARY

[0005] In order to overcome the problems of limited enzymatic effect and high cost of existing fiber-degrading enzyme preparations, the present application provides a compound enzyme preparation for efficiently degrading fiber and a preparation method and application thereof.

[0006] In a first aspect, the present application provides a compound enzyme preparation for efficiently degrading fiber, which adopts the following technical solution:

[0007] A compound enzyme preparation, wherein the compound enzyme preparation is obtained by solid-state fermentation of Aspergillus oryzae, and the compound enzyme preparation comprises xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease;

[0008] The xylanase has an enzyme activity of 7000-10000 U / g, the β-glucanase has an enzyme activity of 500-1000 U / g, the cellulase has an enzyme activity of 4000-7000 U / g, the mannanase has an enzyme activity of 4000-7000 U / g, the pectinase has an enzyme activity of 2000-4000 U / g, the saccharifying enzyme has an enzyme activity of 3000-5000 U / g, and the neutral protease has an enzyme activity of 4000-6000 U / g.

[0009] This application provides a complex enzyme preparation obtained from Aspergillus oryzae through solid-state fermentation. This complex enzyme preparation contains multiple enzymes, including xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease. When used in fiber feed, the synergistic effect of these enzymes degrades cellulose, hemicellulose, and lignin in the fiber feed into oligosaccharides or allows them to be utilized by the animal itself to produce probiotics. This not only improves the palatability of fiber animal feed but also enhances the microbial activity in the animal's digestive tract, promoting the absorption and digestion of nutrients in the feed. Furthermore, the saccharifying enzyme and neutral amylase can effectively degrade anti-nutritional factors in feed ingredients, improving the nutritional value of the feed ingredients, promoting animal growth performance, and ultimately enhancing the economic benefits of the farm.

[0010] In this application, xylanase can effectively degrade hemicellulose, reduce the viscosity of digesta, and improve feed palatability and digestibility; β-glucanase helps to decompose β-glucan, reduce feed viscosity, and improve feed digestibility and absorption efficiency; cellulase can degrade cellulose, promote the utilization of fiber raw materials, and enhance animals' ability to digest feed; mannanase can degrade mannan, improve the physical properties of feed, and reduce the impact of anti-nutritional factors; pectinase can decompose pectin, reduce digesta viscosity, and improve the digestive tract environment; saccharifying enzymes can convert polysaccharides into monosaccharides, improve energy utilization, and increase the nutritional value of feed; neutral protease can decompose proteins to generate small molecule peptides and amino acids, improving the digestibility and absorption rate of protein in feed. This application, through the fermentation of Aspergillus oryzae, obtained a compound enzyme preparation containing the above-mentioned activities. This compound enzyme preparation can reduce the content of cellulose, hemicellulose, and lignin through the synergistic effect of multiple enzymes, significantly improve the degradation efficiency of fiber feed, thereby improving the nutritional value of feed and the digestibility and absorption rate of animals, improving animal growth performance, and greatly increasing the economic benefits of farms. In addition, the above-mentioned compound enzyme preparations are readily available and inexpensive, which can effectively improve the economics of fiber feed.

[0011] Optionally, the solid culture medium used for solid-state fermentation comprises the following components in parts by weight: 5-10 parts wheat bran, 2-4 parts soybean meal, and 0.2-1 parts corn flour.

[0012] In the solid culture medium provided in this application, wheat bran serves as a carbon source, soybean meal as a nitrogen source, and corn flour provides both carbon and nitrogen sources, while also playing a good role in fixation, helping to fix the morphology of the culture medium. Using the above substances as the culture medium in this application can effectively stimulate the mycelial growth of Aspergillus oryzae, thereby producing a large number of non-starch polysaccharide enzymes, saccharifying enzymes, and neutral proteases.

[0013] In some embodiments, the bran may be in the following weight parts: 4-5 parts, 4-8 parts, 4-10 parts, 5-8 parts, 5-10 parts, or 8-10 parts.

[0014] In one specific implementation, the bran may be in the following weight parts: 4 parts, 5 parts, 8 parts, or 10 parts.

[0015] In some implementations, the soybean meal may be in the following weight proportions: 2-3 parts, 2-4 parts, 2-5 parts, 3-4 parts, 3-5 parts, or 4-5 parts.

[0016] In one specific implementation, the soybean meal may be in the following weight proportions: 2 parts, 3 parts, 4 parts, or 5 parts.

[0017] In some embodiments, the corn flour may be in the following weight proportions: 0.2-0.6 parts, 0.2-1 parts, 0.2-1.5 parts, 0.2-5 parts, 0.6-1 parts, 0.6-1.5 parts, 0.6-5 parts, 1-1.5 parts, 1-5 parts, or 1.5-5 parts.

[0018] In one specific implementation, the corn flour may be in the following weight proportions: 0.2 parts, 0.6 parts, 1 part, 1.5 parts, or 5 parts.

[0019] Optionally, the Aspergillus oryzae has the accession number CGMCC No. 40917.

[0020] Secondly, this application provides a method for preparing a compound enzyme preparation, comprising the following steps: preparing a suspension of Aspergillus oryzae spores and solid-state fermentation culture;

[0021] Solid-state fermentation culture: Add Aspergillus oryzae spore suspension to solid culture medium and add sterile water, then ferment at 30-35℃ for 4-7 days; the amount of Aspergillus oryzae spore suspension added is 35-60% of the mass of solid culture medium, and the amount of sterile water added is 10-35% of the mass of solid culture medium.

[0022] This application provides a method for preparing a compound enzyme preparation. This method involves solid-state fermentation of *Aspergillus oryzae*, allowing the microorganisms to grow, reproduce, and metabolize in a low-moisture culture, thereby releasing various non-starch polysaccharide enzymes. These non-starch polysaccharide enzymes can effectively degrade fiber content, improve palatability, enhance feed quality, and facilitate nutrient absorption and digestion in livestock and poultry. Furthermore, the method for preparing the compound enzyme preparation provided in this application has low production costs, high fermentation efficiency, and produces fewer pollutants during solid-state fermentation, making it a very environmentally friendly and green processing method.

[0023] In some embodiments, the amount of the Aspergillus oryzae spore suspension added is 35-41.2%, 35-45.2%, 35-50.0%, 35-58.3%, 41.2-45.2%, 41.2-50.0%, 41.2-58.3%, 45.2-50.0%, 45.2-58.3%, or 50.0-58.3% of the mass of the solid culture medium.

[0024] In one specific implementation, the amount of Aspergillus oryzae spore suspension added is 35%, 41.2%, 45.2%, 50.0%, or 58.3% of the mass of the solid culture medium.

[0025] In some embodiments, the amount of sterile water added is 11.8-17.6%, 11.8-23.5%, 11.8-29.4%, 11.8-35.3%, 17.6-23.5%, 17.6-29.4%, 17.6-35.3%, 23.5-29.4%, 23.5-35.3%, or 29.4-35.3% of the mass of the solid culture medium.

[0026] In one specific implementation, the amount of sterile water added is 11.8%, 17.6%, 23.5%, 29.4%, or 35.3% of the mass of the solid culture medium.

[0027] Optionally, the solid fermentation culture process requires turning the fermented material and adding water. The specific steps are: turning the fermented material once every 19-21 hours, and controlling the water content of the system during the fermentation process to be 35-55%.

[0028] Optionally, after the solid fermentation culture is completed, the moisture content of the fermentation material needs to be adjusted to ≤10%, and then pulverized to obtain a compound enzyme preparation.

[0029] Optionally, the specific steps for preparing the Aspergillus oryzae spore suspension are as follows: Inoculate a single colony of Aspergillus oryzae onto a PDA slant culture medium and incubate at 30-35℃ for 90-100 hours until the spores mature; then wash the mature spores with physiological saline and shake on a shaker for 2 hours; after filtration and resuspending in physiological saline, obtain a final spore concentration of 10. 8-10 9 A suspension of Aspergillus oryzae spores per mL.

[0030] Thirdly, the application of the compound enzyme preparation provided in this application in feed additives or animal feed.

[0031] Fourthly, this application provides an animal feed containing the aforementioned compound enzyme preparation, wherein the amount of the compound enzyme preparation added to the animal feed is 300-900 g / t.

[0032] In summary, this application has the following beneficial effects:

[0033] 1. This application provides a compound enzyme preparation that can be directly added to animal feed as a feed additive. Through the synergistic effect of multiple enzymes, it can efficiently degrade anti-nutritional factors in feed ingredients, improve the digestion and absorption efficiency of feed nutrients by animals, promote intestinal health of animals, improve the body's metabolic level, enhance immunity and disease resistance, improve animal growth performance, and increase the economic benefits of farms.

[0034] 2. The compound enzyme preparation provided in this application is produced by solid-state fermentation of Aspergillus oryzae, so its raw materials are widely available and inexpensive, which can effectively improve the economics of fiber feed. Attached Figure Description

[0035] Figure 1 This is a graph showing the fiber degradation effect of the compound enzyme preparation obtained in Example 2 of this application. Detailed Implementation

[0036] The present application will be further described below with reference to specific embodiments and accompanying drawings. The advantages and features of the present application will become clearer with the description. It should be noted that: unless otherwise specified in the following embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer, and the raw materials used in the following embodiments are all from commercially available sources unless otherwise specified.

[0037] This application provides a compound enzyme preparation, the preparation method of which includes the following steps:

[0038] (1) Preparation of Aspergillus oryzae spore suspension: Inoculate a single colony of Aspergillus oryzae onto a PDA slant medium and incubate at 30-35℃ for 90-100 h until the spores mature; then wash off the mature spores with physiological saline and shake on a shaker for 2 h; after filtration and resuspending in physiological saline, obtain a final spore concentration of 10. 8 -10 9 A suspension of Aspergillus oryzae spores per mL.

[0039] (2) Solid fermentation culture: Add Aspergillus oryzae spore suspension to solid culture medium and add sterile water, then ferment at 30-35℃ for 4-7 days; turn the fermentation mixture every 19-21 hours during the fermentation culture, and test the moisture content of the fermentation mixture after each turning. If the moisture content is lower than 35%, add sterile water to control the moisture content of the system during the fermentation process to 35-55%; after the fermentation culture is completed, the moisture content of the fermentation product needs to be adjusted to ≤10%, and after crushing, a compound enzyme preparation is obtained.

[0040] The solid culture medium used for solid-state fermentation comprises the following components in parts by weight: 5-10 parts wheat bran, 2-4 parts soybean meal, and 0.2-1 parts corn flour; the amount of Aspergillus oryzae spore suspension added is 35-60% of the mass of the solid culture medium, and the amount of sterile water added is 10-35% of the mass of the solid culture medium.

[0041] The composite enzyme preparation obtained in this application includes xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease; the enzyme activity of the xylanase is 7000-10000 U / g, the enzyme activity of the β-glucanase is 500-1000 U / g, the enzyme activity of the cellulase is 4000-7000 U / g, the enzyme activity of the mannanase is 4000-7000 U / g, the enzyme activity of the pectinase is 2000-4000 U / g, the enzyme activity of the saccharifying enzyme is 3000-5000 U / g, and the enzyme activity of the neutral protease is 4000-6000 U / g.

[0042] This application also provides animal feed containing the above-mentioned compound enzyme preparation, wherein the compound enzyme preparation is added to the animal feed at a rate of 300-900 g / t.

[0043] In the embodiments of this application, Aspergillus oryzae is the known strain Aspergillus oryzae PL-N in the prior art, with the accession number CGMCC No.40917; wheat bran, soybean meal and corn flour were all purchased from COFCO Group Co., Ltd.; the raw materials, reagents, solvents and other materials used in this application can all be obtained commercially.

[0044] The present application will be further described in detail below with reference to preparation examples, embodiments, performance testing tests and accompanying drawings.

[0045] Preparation Examples 1-9

[0046] Preparation Examples 1-9 each provide a solid culture medium.

[0047] The difference in the above preparation examples is that the amount of each component added to the solid culture medium is shown in Table 1 below.

[0048] The solid culture medium is prepared as follows: weigh each component as shown in Table 1, then mix them evenly to obtain the solid culture medium.

[0049] Table 1 shows the amount of each component added to the solid culture media provided in Examples 1-9.

[0050]

[0051] Examples 1-9

[0052] Examples 1-9 each provide a compound enzyme preparation.

[0053] The difference between the above embodiments is that the solid culture medium used in Examples 1-9 was derived from Preparation Examples 1-9, respectively.

[0054] The preparation method of the above-mentioned compound enzyme preparation includes the following steps:

[0055] (1) Preparation of Aspergillus oryzae spore suspension: A single colony of Aspergillus oryzae was inoculated onto PDA slant medium and cultured at 30℃ for 100 h until the spores matured; then the mature spores were washed off with physiological saline and shaken on a shaker for 2 h; the spore suspension was filtered through four layers of gauze and resuspended with physiological saline, and the spore concentration was adjusted by counting with a hemocytometer to obtain a spore concentration of 10. 8 A suspension of Aspergillus oryzae spores per mL.

[0056] (2) Solid fermentation culture: Add 70 mL of Aspergillus oryzae spore suspension to 170 g of solid culture medium and add 32 mL of sterile water to obtain a fermentation mixture. Cover with a cloth and then ferment at 30 °C for 6 days. During the fermentation culture, turn the koji once every 20 h. After each turning, test the moisture content of the fermentation mixture. When the moisture content is lower than 35%, add sterile water to make the moisture content of the system 35-55%. After 6 days of fermentation culture, the koji is covered with dark green mycelium. Evaporate the fermentation mixture at 45 °C until the moisture content is ≤10%. After crushing, obtain a compound enzyme preparation.

[0057] Examples 10-17

[0058] Examples 10-17 each provide a compound enzyme preparation.

[0059] The difference between the above embodiment and embodiment 2 is that the amount of solid culture medium, Aspergillus oryzae spore suspension and sterile water added in step (2) is shown in Table 2 below.

[0060] Comparative Examples 1-3

[0061] Comparative Examples 1-3 each provide a compound enzyme preparation.

[0062] The difference between the above comparative example and Example 2 is that the amount of solid culture medium, Aspergillus oryzae spore suspension and sterile water added in step (2) is shown in Table 2 below.

[0063] Table 2. Amounts of solid culture medium, Aspergillus oryzae spore suspension, and sterile water added in Examples 10-17 and Comparative Examples 1-3.

[0064]

[0065] Performance testing

[0066] The activity levels of the compound enzyme preparations obtained in Examples 1-17 and Comparative Examples 1-3 were determined. The enzyme activity determination methods were as follows: xylanase activity was determined according to GB / T 23874-2009; β-glucanase activity was determined according to NY / T 911-2020; cellulase activity was determined according to NY / T 912-2020; mannanase activity was determined according to GB / T 36861-2018; pectinase activity was determined according to QB / T 4482-2013; glucoamylase activity was determined according to QB / T1803-1993; and neutral protease activity was determined according to GB / T 23527-2009. The determination results are shown in Table 3 below.

[0067] Table 3. Enzyme activity assay results of the compound enzyme preparations obtained in Examples 1-17 and Comparative Examples 1-3.

[0068]

[0069] According to the test results in Table 3, the average enzyme activities of xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease in the compound enzyme preparations obtained in Examples 1-17 of this application are 7274-9938 U / g, 547-985 U / g, 4289-6835 U / g, 4125-6973 U / g, 2440-3985 U / g, 3745-4934 U / g, and 4349-5989 U / g, respectively. U / g; while the average enzyme activities of xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease in the complex enzyme preparations obtained in Comparative Examples 1-3 were only 5213-6790 U / g, 320-561 U / g, 2125-4255 U / g, 2049-3942 U / g, 1218-2456 U / g, 2033-3125 U / g, and 3229-4136 U / g, respectively. Therefore, this application demonstrates that by using wheat bran, soybean meal, and corn flour as a solid culture medium, and controlling the addition of Aspergillus oryzae spore suspension to 35-60% of the solid culture medium mass, and the addition of sterile water to 10-35% of the solid culture medium mass, the resulting compound enzyme preparation exhibits high enzyme activity and good degradation effects on fibrous substances and anti-nutritional factors. Its application in fiber feed can improve the nutritional value of feed ingredients and enhance feed utilization.

[0070] The test results of Examples 1-9 showed that the average enzyme activities of xylanase obtained in Examples 1-7 were 8555-9938 U / g (≥8000 U / g), β-glucanase was 789-985 U / g (≥700 U / g), cellulase was 5267-6835 U / g (≥5000 U / g), mannanase was 5342-6973 U / g (≥5000 U / g), pectinase was 3559-3985 U / g (≥3000 U / g), glucoamylase was 4366-4934 U / g (≥4000 U / g), and neutral protease was 5264-5989 U / g (≥5000 U / g). The average enzyme activities of xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease obtained in Examples 8-9 were only 7274-7826 U / g, 547-625 U / g, 4289-4525 U / g, 4125-4343 U / g, 2440-2947 U / g, 3745-4025 U / g, and 4349-4658 U / g, respectively. Therefore, it is evident that by further controlling the addition amounts of wheat bran, soybean meal, and corn flour in the solid culture medium to the following ranges: 5-10 parts wheat bran, 2-4 parts soybean meal, and 0.2-1 parts corn flour, the compound enzyme preparation obtained from fermenting Aspergillus oryzae exhibits higher enzyme activity and better degradation effect on fibrous materials.

[0071] The detection results of Examples 2 and 10-17 showed that the average enzyme activities of xylanase, β-glucanase, cellulase, mannanase, pectinase, and neutral protease obtained in Examples 2, 11-12, and 15-16 were 8627-9938 U / g (≥8000 U / g), 837-985 U / g (≥800 U / g), 5413-6835 U / g (≥5000 U / g), 5667-6973 U / g (≥5000 U / g), 3116-3985 U / g (≥3000 U / g), 4027-4934 U / g (≥4000 U / g), and 5139-5989 U / g (≥5000 U / g). The average enzyme activities of xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme, and neutral protease obtained in Examples 10, 13-14, and 17 were only 7541-8328 U / g, 696-817 U / g, 4732-5028 U / g, 4525-4990 U / g, 2634-2930 U / g, 3273-3951 U / g, and 4535-5025 U / g, respectively. Therefore, it is evident that by further controlling the addition of Aspergillus oryzae spore suspension to 40-50% of the solid culture medium mass and the addition of sterile water to 17-30% of the solid culture medium mass, the resulting composite enzyme preparation exhibits higher enzyme activities and better degradation effects on fibrous materials and anti-nutritional factors.

[0072] Fiber degradation test

[0073] The fiber degradation effect of the compound enzyme preparation obtained in Example 2 was tested. The test method was as follows: 100g of wheat bran was taken, 60g of water and the compound enzyme preparation from Example 2 (added amounts of 0, 0.3g, 0.4g, 0.5g, 0.6g, and 0.7g, respectively) were added, mixed evenly, and then enzymatically hydrolyzed in an incubator at 45℃ for 20h. After enzymatic hydrolysis, the enzyme was immediately inactivated by heating at high temperature for 10min, and then dried and pulverized at 85℃ to obtain the test samples. The total dietary fiber (TDF), IDF (insoluble dietary fiber), SDF (soluble dietary fiber), reducing sugar, water holding capacity, and swelling capacity of each test sample were measured. The determination of TDF, IDF, and SDF of fiber was in accordance with GB / T5009.88-2014, the determination of reducing sugar was performed using the DNS colorimetric method, and the determination of water holding capacity and swelling capacity was in accordance with "Degradation and Functional Evaluation of Dietary Fiber from Bamboo Shoots". The results are as follows. Figure 1 As shown.

[0074] according to Figure 1 The test results show that with the increase of the amount of compound enzyme preparation added, the TDF, IDF, SDF, water holding capacity, and swelling capacity of wheat bran continuously decreased, while the reducing sugar continuously increased. Therefore, this indicates that the compound enzyme preparation provided in this application can degrade fiber and anti-nutritional factors in wheat bran, improve various properties of wheat bran, make it easier for animals to digest and absorb, thereby improving the bioavailability of wheat bran feed, ensuring feeding effects, and reducing feed costs.

[0075] Application Examples 1-3

[0076] Application Examples 1-3 each provide a pig feed.

[0077] The difference in the above application examples lies in the amount of compound enzyme preparations added to pig feed, as shown in Table 4 below.

[0078] The method for preparing pig feed provided in Application Examples 1-3 is as follows: add the compound enzyme preparation provided in Example 2 to ordinary pig feed (purchased from Liaoning Bollet Agricultural and Animal Husbandry Industry Co., Ltd.), mix evenly, and obtain pig feed.

[0079] Comparative Application Examples 1-3

[0080] Comparative application examples 1-3 each provide a pig feed.

[0081] The difference between the above comparative application examples is that the compound enzyme preparations used in pig feed were derived from comparative examples 1-3.

[0082] Pig feeding trial

[0083] Seventy pigs of similar growth condition were selected from a farm as experimental subjects and randomly divided into seven groups of ten pigs each. Three groups were experimental, three were control, and one was a blank control group. The three experimental groups were fed the pig feed used in Examples 1-3; the three control groups were fed the pig feed used in Examples 1-3; and the blank control group was fed ordinary pig feed. The pigs had free access to feed and water and underwent routine immunization and deworming. The experimental period was 40 days. The feed conversion ratio (FCR) of each group was statistically analyzed, and the results are shown in Table 4 below.

[0084] Table 4 Results of the feeding trial on pigs

[0085]

[0086] According to the results in Table 4, when the compound enzyme preparation provided in Example 2 of this application was added to pig feed and fed to pigs for 40 days, the feed conversion ratio (FCR) of the pigs reached 1.51-1.65; while when the compound enzyme preparations provided in Comparative Examples 1-3 were added to pig feed and fed to pigs for 40 days, the FCR of the pigs was 1.71-1.78; and when pigs were fed ordinary pig feed for 40 days, the FCR of the pigs was as high as 1.83. Therefore, it is demonstrated that the compound enzyme preparation provided in this application can degrade fiber and anti-nutritional factors in pig feed, improve the nutritional value of feed ingredients, significantly reduce the FCR of pig farming, and improve farming efficiency.

[0087] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A compound enzyme preparation, characterized in that, The compound enzyme preparation is obtained by Aspergillus oryzae through solid-state fermentation, and the compound enzyme preparation includes xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease. The xylanase has an enzyme activity of 8000-10000 U / g, the β-glucanase has an enzyme activity of 700-1000 U / g, the cellulase has an enzyme activity of 5000-7000 U / g, the mannanase has an enzyme activity of 5000-7000 U / g, the pectinase has an enzyme activity of 3000-4000 U / g, the saccharifying enzyme has an enzyme activity of 4000-5000 U / g, and the neutral protease has an enzyme activity of 5000-6000 U / g. The solid culture medium used for solid-state fermentation comprises the following components in parts by weight: 5-10 parts wheat bran, 2-4 parts soybean meal, and 0.2-1 parts corn flour; The preparation method of the compound enzyme preparation includes the following steps: preparing Aspergillus oryzae spore suspension and solid fermentation culture; Solid-state fermentation culture: Add Aspergillus oryzae spore suspension to a solid culture medium and replenish with sterile water, then ferment at 30-35℃ for 4-7 days; the amount of Aspergillus oryzae spore suspension added is 40-50% of the mass of the solid culture medium, and the amount of sterile water added is 17-30% of the mass of the solid culture medium; the final spore concentration of the Aspergillus oryzae spore suspension is 10. 8 -10 9 cells / mL; The preservation number of the Aspergillus oryzae is CGMCC No. 40917.

2. The compound enzyme preparation according to claim 1, characterized in that, The solid fermentation process requires turning the fermented material and adding water. The specific steps are as follows: turn the fermented material once every 19-21 hours, and control the water content of the system to 35-55% during the fermentation process.

3. The compound enzyme preparation according to claim 2, characterized in that, After the solid-state fermentation culture is completed, the moisture content of the fermentation material needs to be adjusted to ≤10%, and then pulverized to obtain a compound enzyme preparation.

4. The compound enzyme preparation according to claim 1, characterized in that, The specific steps for preparing the Aspergillus oryzae spore suspension are as follows: A single colony of Aspergillus oryzae is inoculated onto a PDA slant culture medium and cultured at 30-35℃ for 90-100 hours until the spores mature; then the mature spores are washed off with physiological saline and shaken on a shaker for 2 hours; after filtration and resuspending in physiological saline, a final spore concentration of 10 is obtained. 8 -10 9 A suspension of Aspergillus oryzae spores per mL.

5. The use of the compound enzyme preparation as described in any one of claims 1-4 in feed additives or animal feed.

6. An animal feed comprising the compound enzyme preparation according to any one of claims 1-4, characterized in that, The compound enzyme preparation is added to animal feed at a rate of 300-900 g / t.

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