Compound enzyme preparation for efficiently degrading fibers as well as preparation method and application of compound enzyme preparation

Abstract

The invention relates to the technical field of microbial fermentation, and particularly discloses a compound enzyme preparation for efficiently degrading fibers as well as a preparation method and application of the compound enzyme preparation. The compound enzyme preparation provided by the invention is obtained by solid state fermentation of aspergillus oryzae, and the compound enzyme preparation comprises xylanase, beta-glucanase, cellulase, mannase, pectinase, saccharifying enzyme and neutral protease. Furthermore, a solid culture medium used for solid-state fermentation comprises the following components in parts by weight: 5-10 parts of bran, 2-4 parts of soybean meal and 0.2-1 part of corn flour; the invention also provides a preparation method of the compound enzyme preparation. The preparation method comprises the following steps: preparing an aspergillus oryzae spore suspension and performing solid fermentation culture. The compound enzyme preparation provided by the invention has a remarkable degradation effect on fibers, and when the compound enzyme preparation is applied to animal feed, the digestibility of animals to the fiber feed can be remarkably improved, the growth performance and the health condition of the animals are improved, the breeding cost is reduced, and the economic benefit of a breeding farm is improved.

Description

Technical Field

[0001] The present application relates to the field of microbial fermentation technology, and in particular to a composite enzyme preparation for efficiently degrading fiber, and a preparation method and application thereof. Background Art

[0002] With the rapid development of animal husbandry, the problem of feed shortage in my country is becoming increasingly serious. Since high-energy grains are mainly used for human consumption, more high-fiber grain processing byproducts such as corn stalks are used in animal feed. These agricultural byproducts are rich in organic matter and are considered to be important renewable resources. However, the fiber substances contained in these byproducts pose certain challenges to animal digestion, so it is difficult to fully realize the economic value of agricultural byproducts as animal feed.

[0003] At present, in order to improve the utilization rate of high-fiber feed, the industry mainly uses physical, chemical and biological methods to process fiber feed. Among them, the physical method is to change the fiber structure through heat treatment, high-pressure cooking and other methods to improve the enzymatic hydrolysis efficiency of the fiber, making it easy for animals to absorb; however, this method has a very limited effect on the fiber and requires a lot of energy consumption, so it cannot be applied on a large scale. The chemical method is to use acid and alkali solutions to destroy the chemical bonds between fiber molecules and improve the degradation efficiency of the fiber, but the use of chemical reagents will increase the processing cost and cause pollution to the environment. The biological method is to use the enzymatic hydrolysis of enzyme preparations to decompose complex fiber components into small molecules that are easier for animals to absorb. It has low operating costs and little pollution to the environment. It is currently the most widely used fiber feed processing method.

[0004] However, although the biological method has reduced the processing cost of feed fiber to a certain extent, it still has the following problems: first, the effect of a single enzyme in the enzymatic hydrolysis process is limited, and it is difficult to effectively deal with the complex structure of the fiber, resulting in low enzymatic hydrolysis efficiency; second, the existing commercial enzyme preparations are expensive, which limits their large-scale application. Therefore, if the enzyme can be produced and used locally, the development of an efficient and low-cost composite enzyme preparation is of great significance to improving the economic efficiency of fiber feed. Summary of the invention

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

[0006] In the first aspect, the present application provides a composite enzyme preparation for efficiently degrading fibers, using the following technical solution: A composite enzyme preparation, which is obtained by solid-state fermentation of Aspergillus oryzae, and comprises xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease; The enzyme activity of the xylanase is 7000-10000U / g, the enzyme activity of the β-glucanase is 500-1000U / g, the enzyme activity of the cellulase is 4000-7000U / g, the enzyme activity of the mannanase is 4000-7000U / g, the enzyme activity of the pectinase is 2000-4000U / g, the enzyme activity of the saccharifying enzyme is 3000-5000U / g, and the enzyme activity of the neutral protease is 4000-6000U / g.

[0007] The present application provides a composite enzyme preparation obtained by solid-state fermentation of Aspergillus oryzae, which comprises a variety of enzymes such as xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease. The composite enzyme preparation is used in fiber feed, and the cellulose, hemicellulose and lignin in the fiber feed can be degraded into oligosaccharides or utilized by the synergistic action of a variety of enzymes to produce beneficial biomass, which can not only improve the palatability of fiber animal feed, but also improve the microbial activity in the animal digestive tract and promote the absorption and digestion of nutrients in the feed by the animal; in addition, saccharifying enzyme and neutral amylase can also effectively degrade anti-nutritional factors in feed raw materials, improve the nutritional value of feed raw materials, promote the growth performance of animals, and thus improve the economic benefits of farms.

[0008] In this application, xylanase can effectively degrade hemicellulose, reduce the viscosity of chyme, and improve the palatability and digestibility of feed; β-glucanase helps to decompose β-glucan, reduce feed viscosity, and improve the digestion and absorption efficiency of feed; cellulase can degrade cellulose, promote the utilization of fiber raw materials, and enhance the digestion ability of animals to feed; mannanase can degrade mannan, improve the physical properties of feed, and reduce the impact of anti-nutritional factors; pectinase can decompose pectin, reduce the viscosity of chyme, and improve the digestive tract environment; saccharifying enzyme can convert polysaccharides into monosaccharides, improve energy utilization, and increase the nutritional value of feed; neutral protease can decompose protein, generate small molecule peptides and amino acids, and improve the digestion and absorption rate of protein in feed. This application obtains a composite enzyme preparation containing the above-mentioned vitality by fermenting Aspergillus oryzae. The composite enzyme preparation can reduce the content of cellulose, hemicellulose and lignin through the synergistic action of multiple enzymes, significantly improve the degradation efficiency of fiber feed, thereby improving the nutritional value of feed and the digestion and absorption rate of animals, improving the growth performance of animals, and greatly increasing the economic benefits of farms. In addition, the above-mentioned complex enzyme preparation is convenient to source and low in cost, and can effectively improve the economy of fiber feed.

[0009] Optionally, the solid culture medium used for the solid-state fermentation comprises the following components in parts by weight: 5-10 parts of bran, 2-4 parts of soybean meal and 0.2-1 part of corn flour.

[0010] In the solid culture medium provided in the present application, bran serves as a carbon source, soybean meal serves as a nitrogen source, and corn flour can provide both a carbon source and a nitrogen source, and also plays a good fixation role, helping to fix the morphology of the culture medium; the present application uses the above substances as a culture medium, which can effectively stimulate the mycelial growth of Aspergillus oryzae, thereby producing a large amount of non-starch polysaccharide enzymes, saccharification enzymes and neutral proteases.

[0011] In some embodiments, the bran may be present in an amount of 4-5 parts, 4-8 parts, 4-10 parts, 5-8 parts, 5-10 parts, or 8-10 parts by weight.

[0012] In a specific embodiment, the weight portion of the bran can be 4 parts, 5 parts, 8 parts or 10 parts.

[0013] In some embodiments, the soybean meal may be present in an amount of 2-3 parts, 2-4 parts, 2-5 parts, 3-4 parts, 3-5 parts, or 4-5 parts by weight.

[0014] In a specific embodiment, the weight proportions of the soybean meal can be 2 parts, 3 parts, 4 parts, or 5 parts.

[0015] In some embodiments, the weight portion of the corn flour can be 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.

[0016] In a specific embodiment, the weight portion of the corn flour can be 0.2 parts, 0.6 parts, 1 parts, 1.5 parts or 5 parts.

[0017] Optionally, the Aspergillus oryzae is deposited with CGMCC No.40917.

[0018] In a second aspect, the present application provides a method for preparing a composite enzyme preparation, comprising the following steps: preparing a suspension of Aspergillus oryzae spores and solid fermentation culture; Solid fermentation culture: add Aspergillus oryzae spore suspension to the solid culture medium, and add sterile water, and then ferment and culture at 30-35°C for 4-7 days; the addition amount of the Aspergillus oryzae spore suspension is 35-60% of the mass of the solid culture medium, and the addition amount of the sterile water is 10-35% of the mass of the solid culture medium.

[0019] The present application provides a method for preparing a composite enzyme preparation, which allows the microorganism to grow, reproduce and metabolize in a culture medium with low moisture content by solid-state fermentation of Aspergillus oryzae, thereby releasing a variety of non-starch polysaccharide enzymes, which can effectively degrade fiber content, improve palatability, improve feed quality, and facilitate nutrient absorption and digestion of livestock and poultry. In addition, the preparation method of the composite enzyme preparation provided by the present application has low production cost, high fermentation efficiency, and less pollutants produced by solid-state fermentation, which is very friendly to the environment and is a green and environmentally friendly processing method.

[0020] In some embodiments, the Aspergillus oryzae spore suspension is added in an amount of 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.

[0021] In a specific embodiment, the added amount of the Aspergillus oryzae spore suspension is 35%, 41.2%, 45.2%, 50.0% or 58.3% of the mass of the solid culture medium.

[0022] 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.

[0023] In a specific embodiment, 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.

[0024] Optionally, during the solid fermentation culture process, the fermented material needs to be turned over and hydrated, and the specific steps are: turning over once every 19-21 hours, and controlling the water content of the system to 35-55% during the fermentation process.

[0025] Optionally, after the solid fermentation culture is completed, the water content of the fermented product needs to be adjusted to ≤10%, and the product is crushed to obtain a composite enzyme preparation.

[0026] Optionally, the specific steps of preparing the Aspergillus oryzae spore suspension are: inoculating a single Aspergillus oryzae colony into a PDA slant medium, culturing at 30-35° C. for 90-100 h until the spores mature; then washing the mature spores with physiological saline, and shaking on a shaking table for 2 h; filtering, and resuspending with physiological saline to obtain a final spore concentration of 10 8-10 9 / mL of Aspergillus oryzae spore suspension.

[0027] In a third aspect, the present application provides an application of the complex enzyme preparation in feed additives or animal feed.

[0028] In a fourth aspect, the present application provides an animal feed containing the complex enzyme preparation, wherein the addition amount of the complex enzyme preparation in the animal feed is 300-900 g / t.

[0029] In summary, this application has the following beneficial effects: 1. The present application provides a complex enzyme preparation, which can be directly added to animal feed as a feed additive. Through the synergistic action of multiple enzymes, it can efficiently degrade anti-nutritional factors in feed raw materials, improve the digestion and absorption efficiency of feed nutrients by animals, promote animal intestinal health, improve the body's metabolic level, enhance immunity and disease resistance, improve animal growth performance, and increase the economic benefits of farms.

[0030] 2. The composite enzyme preparation provided in the present application is prepared by solid-state fermentation of Aspergillus oryzae, so its raw material source is wide and the cost is low, which can effectively improve the economy of fiber feed. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a diagram of the fiber degradation effect of the composite enzyme preparation obtained in Example 2 of the present application. DETAILED DESCRIPTION

[0032] The present application is further described below in conjunction with specific examples and drawings, and the advantages and features of the present application will become clearer as the description proceeds. It is particularly noted that if no specific conditions are specified in the following examples, the conventional conditions or the conditions recommended by the manufacturer are followed, and the raw materials used in the following examples can all be sourced from common commercial sources unless otherwise specified.

[0033] The present application provides a composite enzyme preparation, and a method for preparing the composite enzyme preparation comprises the following steps: (1) Preparation of Aspergillus oryzae spore suspension: Inoculate a single colony of Aspergillus oryzae on a PDA slant medium and culture at 30-35°C for 90-100 h until the spores mature; then wash the mature spores with saline and shake on a shaker for 2 h; filter and resuspend in saline to obtain a final spore concentration of 10 8 -10 9 / mL of Aspergillus oryzae spore suspension.

[0034] (2) Solid fermentation culture: Add Aspergillus oryzae spore suspension to the solid culture medium, and add sterile water, and then ferment and culture at 30-35°C for 4-7 days; during the fermentation culture, turn the koji every 19-21 hours, and test the water content of the fermentation mixture after each turning. When the water content is lower than 35%, add sterile water, and control the water content of the system to 35-55% during the fermentation process; after the fermentation culture is completed, the water content of the fermented product needs to be adjusted to ≤10%, and then crushed to obtain a composite enzyme preparation; The solid culture medium used for the solid-state fermentation comprises the following components in parts by weight: 5-10 parts of bran, 2-4 parts of soybean meal and 0.2-1 part of corn flour; the addition amount of the Aspergillus oryzae spore suspension is 35-60% of the mass of the solid culture medium, and the addition amount of the sterile water is 10-35% of the mass of the solid culture medium.

[0035] The composite enzyme preparation prepared in the present application includes xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease; the enzyme activity of the xylanase is 7000-10000U / g, the enzyme activity of the β-glucanase is 500-1000U / g, the enzyme activity of the cellulase is 4000-7000U / g, the enzyme activity of the mannanase is 4000-7000U / g, the enzyme activity of the pectinase is 2000-4000U / g, the enzyme activity of the saccharifying enzyme is 3000-5000U / g, and the enzyme activity of the neutral protease is 4000-6000U / g.

[0036] The present application also provides animal feed comprising the above-mentioned complex enzyme preparation, wherein the addition amount of the complex enzyme preparation in the animal feed is 300-900 g / t.

[0037] In the examples of the present application, Aspergillus oryzae is a known strain of Aspergillus oryzae PL-N in the prior art, with a preservation number of CGMCC No.40917; bran, soybean meal and corn flour were all purchased from COFCO Corporation; the raw materials, reagents, solvents, etc. used in the present application can all be obtained commercially.

[0038] The present application is further described in detail below in combination with preparation examples, embodiments, performance testing experiments and accompanying drawings. Preparation Example 1-9

[0039] Preparation Examples 1-9 each provide a solid culture medium.

[0040] The difference between the above preparation examples is that the addition amount of each component in the solid culture medium is shown in Table 1 below.

[0041] The preparation method of the solid culture medium is as follows: weigh the components as shown in Table 1, and then mix them evenly to obtain the solid culture medium.

[0042] Table 1 Addition amount of each component in the solid culture medium provided in Preparation Examples 1-9 Examples 1-9

[0043] Examples 1-9 respectively provide a composite enzyme preparation.

[0044] The difference between the above embodiments is that the solid culture medium used in Embodiments 1-9 is derived from Preparation Examples 1-9, respectively.

[0045] The preparation method of the above-mentioned complex enzyme preparation comprises the following steps: (1) Preparation of Aspergillus oryzae spore suspension: A single colony of Aspergillus oryzae was inoculated into a PDA slant medium and cultured at 30°C for 100 h until the spores matured; the mature spores were then washed off with saline and shaken for 2 h; the spore suspension was filtered through four layers of gauze and resuspended with saline, and the spore concentration was adjusted by counting with a hemocytometer to obtain a spore concentration of 10 8 / mL of Aspergillus oryzae spore suspension.

[0046] (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, which is covered with a cloth; then ferment and culture at 30°C for 6 days; during the fermentation culture, turn the koji every 20 hours, and measure the water content of the fermentation mixture after each turning. When the water content is lower than 35%, add sterile water to make the water content of the system 35-55%; after 6 days of fermentation, the koji is covered with dark green hyphae, and the fermentation mixture is evaporated at 45°C to a water content of ≤10%, and then pulverized to obtain a composite enzyme preparation. Examples 10-17

[0047] Examples 10-17 each provide a composite enzyme preparation.

[0048] The difference between the above embodiment and embodiment 2 is that in step (2), the added amounts of solid culture medium, Aspergillus oryzae spore suspension and sterile water are as shown in Table 2 below. Comparative Examples 1-3

[0049] Comparative Examples 1-3 each provide a composite enzyme preparation.

[0050] The difference between the comparative example and Example 2 is that in step (2), the amounts of solid culture medium, Aspergillus oryzae spore suspension and sterile water added are as shown in Table 2 below.

[0051] Table 2 Addition amount of solid culture medium, Aspergillus oryzae spore suspension and sterile water in Examples 10-17 and Comparative Examples 1-3 Performance testing

[0052] The composite enzyme preparations obtained in Examples 1-17 and Comparative Examples 1-3 were subjected to various enzyme activity index determinations. The enzyme activity determination methods of the enzymes were as follows: the enzyme activity determination of xylanase was based on GB / T 23874-2009, the enzyme activity determination of β-glucanase was based on NY / T 911-2020, the enzyme activity determination of cellulase was based on NY / T 912-2020, the enzyme activity determination of mannanase was based on GB / T 36861-2018, the enzyme activity determination of pectinase was based on QB / T 4482-2013, the enzyme activity determination of saccharifying enzyme was based on QB / T1803-1993, and the enzyme activity determination of neutral protease was based on GB / T 23527-2009. The determination results are shown in Table 3 below.

[0053] Table 3 Enzyme activity test results of composite enzyme preparations obtained in Examples 1-17 and Comparative Examples 1-3

[0054] According to the test results in Table 3, the average enzyme activity of xylanase in the composite enzyme preparations obtained in Examples 1-17 of the present application is 7274-9938 U / g, the average enzyme activity of β-glucanase is 547-985 U / g, the average enzyme activity of cellulase is 4289-6835 U / g, the average enzyme activity of mannanase is 4125-6973 U / g, the average enzyme activity of pectinase is 2440-3985 U / g, the average enzyme activity of saccharifying enzyme is 3745-4934 U / g, and the average enzyme activity of neutral protease is 4349-5989 U / g; while the average enzyme activity of xylanase in the composite enzyme preparations obtained in Comparative Examples 1-3 was only 5213-6790U / g, the average enzyme activity of β-glucanase was only 320-561U / g, the average enzyme activity of cellulase was only 2125-4255U / g, the average enzyme activity of mannanase was only 2049-3942U / g, the average enzyme activity of pectinase was only 1218-2456U / g, the average enzyme activity of saccharifying enzyme was only 2033-3125U / g, and the average enzyme activity of neutral protease was only 3229-4136U / g. Therefore, it is explained that the present application adopts bran, soybean meal and corn flour as solid culture medium, and controls the addition amount of Aspergillus oryzae spore suspension to 35-60% of the mass of the solid culture medium, and the addition amount of sterile water to 10-35% of the mass of the solid culture medium. The enzymes in the prepared composite enzyme preparation have high activity and good degradation effect on fiber substances and anti-nutritional factors. Using it in fiber feed can improve the nutritional value of feed raw materials and improve feed utilization.

[0055] The test results of Examples 1-9 show that the average enzyme activity of the xylanase obtained in Example 1-7 is 8555-9938 U / g (≥8000 U / g), the average enzyme activity of the β-glucanase is 789-985 U / g (≥700 U / g), the average enzyme activity of the cellulase is 5267-6835 U / g (≥5000 U / g), the average enzyme activity of the mannanase is 5342-6973 U / g (≥5000 U / g), the average enzyme activity of the pectinase is 3559-3985 U / g (≥3000 U / g), the average enzyme activity of the saccharifying enzyme is 4366-4934 U / g (≥4000 U / g), and the average enzyme activity of the neutral protease is 5264-5989 U / g (≥5000 U / g); while the average enzyme activity of xylanase obtained in Examples 8-9 is only 7274-7826U / g, the average enzyme activity of β-glucanase is only 547-625U / g, the average enzyme activity of cellulase is only 4289-4525U / g, the average enzyme activity of mannanase is only 4125-4343U / g, the average enzyme activity of pectinase is only 2440-2947U / g, the average enzyme activity of saccharifying enzyme is only 3745-4025U / g, and the average enzyme activity of neutral protease is only 4349-4658U / g. Therefore, it is explained that the application further controls the addition amount of bran, soybean meal and corn flour in the solid culture medium within the following range: 5-10 parts of bran, 2-4 parts of soybean meal and 0.2-1 parts of corn flour, and the enzyme activity of the composite enzyme preparation obtained by fermenting Aspergillus oryzae is higher, and its degradation effect on fiber substances is better.

[0056] The test results of Example 2 and Examples 10-17 show that the average enzyme activity of the xylanase obtained in Example 2, Examples 11-12, and Examples 15-16 is 8627-9938 U / g (≥8000 U / g), the average enzyme activity of the β-glucanase is 837-985 U / g (≥800 U / g), the average enzyme activity of the cellulase is 5413-6835 U / g (≥5000 U / g), the average enzyme activity of the mannanase is 5667-6973 U / g (≥5000 U / g), the average enzyme activity of the pectinase is 3116-3985 U / g (≥3000 U / g), the average enzyme activity of the saccharifying enzyme is 4027-4934 U / g (≥4000 U / g), and the average enzyme activity of the neutral protease is 5139-5989 U / g (≥5000 U / g); while the average enzyme activity of xylanase obtained in Example 10, Example 13-14, and Example 17 is only 7541-8328U / g, the average enzyme activity of β-glucanase is only 696-817U / g, the average enzyme activity of cellulase is only 4732-5028U / g, the average enzyme activity of mannanase is only 4525-4990U / g, the average enzyme activity of pectinase is only 2634-2930U / g, the average enzyme activity of saccharifying enzyme is only 3273-3951U / g, and the average enzyme activity of neutral protease is only 4535-5025U / g. Therefore, it is explained that the application further controls the addition amount of Aspergillus oryzae spore suspension to 40-50% of the mass of the solid culture medium, and the addition amount of sterile water is controlled to 17-30% of the mass of the solid culture medium, and the enzyme activity of each enzyme in the prepared composite enzyme preparation is higher, and the degradation effect of fiber substances and anti-nutritional factors is better. Fiber degradation test

[0057] The fiber degradation effect of the composite enzyme preparation obtained in Example 2 was tested. The test method was as follows: take 100g of wheat bran, add 60g of water and the composite enzyme preparation of Example 2 (the added amounts were 0, 0.3g, 0.4g, 0.5g, 0.6g, and 0.7g, respectively), mix well, and then enzymolyze in a 45°C incubator for 20h; after the enzymolysis is completed, immediately place it in a high temperature for 10min to heat and inactivate the enzyme, dry and crush at 85°C to obtain the test sample. The total dietary fiber (TDF), IDF (insoluble dietary fiber), SDF (water-soluble dietary fiber), reducing sugar, water holding capacity and swelling capacity of each test sample were tested respectively. The determination of TDF, IDF, and SDF of fiber refers to GB / T5009.88-2014, the determination of reducing sugar uses the DNS colorimetric method, and the determination of water holding capacity and swelling capacity refers to "Degradation and Functional Evaluation of Dietary Fiber of Bamboo Shoots". The results are as follows Figure 1 shown.

[0058] according to Figure 1 The test results show that with the increase in the amount of compound enzyme preparation added, the TDF, IDF, SDF, water holding capacity and swelling power of wheat bran continue to decrease, and reducing sugar continues to increase. Therefore, it is shown that the compound enzyme preparation provided by the present application can degrade the fiber and anti-nutritional factors in wheat bran, improve the various properties of wheat bran, make it easy for animals to digest and absorb, and then can improve the bioavailability of wheat bran feed, ensure feeding effect, and reduce feed costs. Application Examples 1-3

[0059] Application Examples 1-3 each provide a kind of pig feed.

[0060] The difference between the above application examples is that the addition amount of the complex enzyme preparation used in the pig feed is shown in Table 4 below.

[0061] The preparation method of the pig feed provided in Application Examples 1-3 is as follows: add the complex enzyme preparation provided in Example 2 to ordinary pig feed (purchased from Liaoning Bollet Agriculture and Animal Husbandry Industry Co., Ltd.), mix well, and obtain pig feed. Comparative Application Examples 1-3

[0062] Comparative Application Examples 1-3 each provide a kind of pig feed.

[0063] The difference between the above comparative application examples is that the complex enzyme preparations used in the pig feed are derived from comparative examples 1-3 respectively. Pig feeding trial

[0064] 70 pigs of the same growth condition were selected from the farm as the test subjects and randomly divided into 7 groups, each with 10 pigs; 3 groups were experimental groups, 3 groups were comparison groups, and 1 group was a blank control group; the 3 experimental groups were fed with the pig feeds of Application Examples 1-3; the 3 comparison groups were fed with the pig feeds of Comparative Application Examples 1-3, and the 1 blank control group was fed with ordinary pig feed. The pigs were allowed to eat and drink freely, and were immunized and dewormed as usual. The test period was 40 days. The feed-to-meat ratio of the pigs in each group was statistically analyzed, and the results are shown in Table 4 below.

[0065] Table 4 Results of pig feeding experiment

[0066] According to the results in Table 4, when the composite enzyme preparation provided in Example 2 of the present application is added to pig feed and fed to pigs for 40 days, the feed-to-meat ratio of the pigs can reach 1.51-1.65; when the composite enzyme preparation provided in Comparative Examples 1-3 is added to pig feed and fed to pigs for 40 days, the feed-to-meat ratio of the pigs can be 1.71-1.78; when ordinary pig feed is used to feed pigs for 40 days, the feed-to-meat ratio of the pigs is as high as 1.83. Therefore, it is shown that the composite enzyme preparation provided in the present application can degrade the fiber substances and anti-nutritional factors in pig feed, improve the nutritional value of feed raw materials, significantly reduce the feed-to-meat ratio of pig farming, and improve the farming efficiency.

[0067] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A composite enzyme preparation, characterized in that: The composite enzyme preparation is obtained by solid-state fermentation of Aspergillus oryzae, and the composite enzyme preparation comprises xylanase, β-glucanase, cellulase, mannanase, pectinase, saccharifying enzyme and neutral protease; The enzyme activity of the xylanase is 7000-10000U / g, the enzyme activity of the β-glucanase is 500-1000U / g, the enzyme activity of the cellulase is 4000-7000U / g, the enzyme activity of the mannanase is 4000-7000U / g, the enzyme activity of the pectinase is 2000-4000U / g, the enzyme activity of the saccharifying enzyme is 3000-5000U / g, and the enzyme activity of the neutral protease is 4000-6000U / g.

2. The complex enzyme preparation according to claim 1, characterized in that The solid culture medium used for the solid-state fermentation comprises the following components in parts by weight: 5-10 parts of bran, 2-4 parts of soybean meal and 0.2-1 part of corn flour.

3. The complex enzyme preparation according to claim 1, characterized in that The deposit number of the Aspergillus oryzae is CGMCC No.40917.

4. The method for preparing the complex enzyme preparation according to any one of claims 1 to 3, characterized in that: The following steps are involved: Preparation of Aspergillus oryzae spore suspension and solid fermentation culture; Solid fermentation culture: add Aspergillus oryzae spore suspension to the solid culture medium, and add sterile water, and then ferment and culture at 30-35°C for 4-7 days; the addition amount of the Aspergillus oryzae spore suspension is 35-60% of the mass of the solid culture medium, and the addition amount of the sterile water is 10-35% of the mass of the solid culture medium.

5. The method for preparing the complex enzyme preparation according to claim 4, characterized in that: During the solid fermentation culture process, the fermented material needs to be turned over and hydrated. The specific steps are: turning over once every 19-21 hours, and controlling the water content of the system to 35-55% during the fermentation process.

6. The method for preparing the complex enzyme preparation according to claim 5, characterized in that: After the solid fermentation culture is completed, the water content of the fermented product needs to be adjusted to ≤10%, and the product is crushed to obtain a composite enzyme preparation.

7. The method for preparing the complex enzyme preparation according to claim 4, characterized in that: The specific steps of preparing the Aspergillus oryzae spore suspension are: inoculating a single Aspergillus oryzae colony into a PDA slant medium, culturing at 30-35° C. for 90-100 hours until the spores mature; then washing the mature spores with physiological saline, and shaking on a shaking table for 2 hours; filtering, and resuspending with physiological saline to obtain a final spore concentration of 10 8 -10 9 / mL of Aspergillus oryzae spore suspension.

8. Use of the complex enzyme preparation according to any one of claims 1 to 3 and the complex enzyme preparation obtained by the preparation method according to any one of claims 4 to 7 in feed additives or animal feed.

9. An animal feed comprising the complex enzyme preparation according to any one of claims 1 to 3 or the complex enzyme preparation obtained by the preparation method according to any one of claims 4 to 7, characterized in that: The compound enzyme preparation is added in an amount of 300-900 g / t in animal feed.

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