Compound enzyme preparation of aspergillus niger fermented traditional Chinese medicine as well as preparation method and application of compound enzyme preparation

Abstract

The invention relates to the technical field of traditional Chinese medicine fermentation, and particularly discloses a compound enzyme preparation of aspergillus niger fermented traditional Chinese medicine as well as a preparation method and application of the compound enzyme preparation. The invention provides a compound enzyme preparation of aspergillus niger fermented traditional Chinese medicines. The compound enzyme preparation is prepared by fermenting a mixture of basic nutrients and the traditional Chinese medicines by using the aspergillus niger, the weight ratio of the aspergillus niger to the basic nutrients to the traditional Chinese medicines is 1: (150-180): (50-70); the traditional Chinese medicine consists of the following components in parts by weight: 22-32 parts of coptis chinensis, 17-27 parts of honeysuckle, 12-22 parts of dandelion, 8-14 parts of agastache rugosus, 4-9 parts of endive, 4-9 parts of hawthorn and 4-9 parts of dark plum. The compound enzyme preparation provided by the invention is high in enzyme activity and wide in enzyme system, can effectively degrade multi-component anti-nutritional factors and improve the feed utilization rate, and can be used as a livestock feed additive to be applied to feed, so that the immunity can be improved, and the intestinal health can be regulated.

Description

Technical Field

[0001] The present application relates to the technical field of Chinese medicine fermentation, and in particular to a composite enzyme preparation of Chinese medicine fermented by Aspergillus niger, and a preparation method and application thereof. Background Art

[0002] Aspergillus niger, belonging to the subphylum Aspergillus, class Hyphomycetes, order Hyphomycetales, family Hyphomycetaceae, is a common non-toxic Aspergillus fungus. Aspergillus niger has a strong ability to express foreign genes and efficient protein expression, secretion and modification, and the recombinant has high genetic stability. Therefore, Aspergillus niger has become an important enzyme expression system and an important industrial enzyme production strain. It is reported that Aspergillus niger can produce a variety of enzymes such as amylase, saccharifying enzyme, protease, cellulase, xylanase, pectinase, lipase, glucose oxidase, etc.

[0003] After the Chinese medicine is fermented by microorganisms, the plant cell wall is enzymatically hydrolyzed, and active substances such as alkaloids, organic acids, polysaccharides, flavonoids, and total phenols are dissolved in large quantities. Alkaloids, as the main active ingredients of Coptis chinensis, have strong antibacterial effects, can improve immunity, and protect the gastric mucosa. Organic acids are natural molecular substances with antibacterial activity and are the most effective non-drug substitutes for feed antibiotics. They can be single acids or mixtures of multiple acids. In addition to their antibiotic effects, organic acids also have several other effects, including lowering the pH of digesta and increasing pancreatic secretion. These organic acids have been used in the diet of weaned piglets for a long time, and have a positive effect on the health and growth of piglets, improving the growth and development of pig immune organs, reducing morbidity, and regulating their gastrointestinal function. Polysaccharides are high-molecular compounds with complex structures and special activities. They have many aspects of biological activity and are biological non-specific immunostimulants. Flavonoids have antioxidant, anti-inflammatory, and hypoglycemic effects, which can promote the redistribution of nutrients in animals, improve protein synthesis efficiency, and improve breeding benefits. Phenolic substances have antioxidant effects, which help reduce oxidative stress in animals, improve their immunity and health, and also have antibacterial effects and improve meat quality.

[0004] However, there are few systematic studies and reports on the fermentation methods of traditional Chinese medicine using Aspergillus niger. Summary of the invention

[0005] In order to solve the above technical problems, the present application provides a composite enzyme preparation of Chinese medicine fermented by Aspergillus niger and a preparation method and application thereof.

[0006] In a first aspect, the present application provides a composite enzyme preparation of Chinese medicine fermented by Aspergillus niger, characterized in that the composite enzyme preparation is prepared by fermenting a mixture of basic nutrients and Chinese medicine by Aspergillus niger; The weight ratio of the Aspergillus niger, the basic nutrients and the traditional Chinese medicine is 1:150-180:50-70; The traditional Chinese medicine consists of the following components in parts by weight: 22-32 parts of coptis root, 17-27 parts of honeysuckle, 12-22 parts of dandelion, 8-14 parts of patchouli, 4-9 parts of chicory, 4-9 parts of hawthorn and 4-9 parts of ebony.

[0007] By adopting the above-mentioned technical scheme, during the fermentation of traditional Chinese medicine by Aspergillus niger, the plant cell walls are enzymatically hydrolyzed, and active substances such as alkaloids, organic acids, polysaccharides, flavonoids, total phenols, etc. are dissolved in large quantities, thereby effectively improving the utilization rate of traditional Chinese medicine and the palatability of feed. The complex enzyme preparation can also quickly and effectively improve the immunity of livestock and resist viruses.

[0008] In the technical scheme provided in the present application, each raw material component in traditional Chinese medicine plays a vital role: Coptis chinensis contains alkaloids such as berberine, coptisine, methyl coptisine, palmatine, jatrorrhizine, epicoptisine, etc., which have the effects of anti-tumor, antibacterial, lowering blood lipids, lowering blood sugar and promoting callus formation; Honeysuckle contains pharmacologically active ingredients such as chlorogenic acid and luteolin glycosides, which have strong inhibitory effects on various pathogenic bacteria such as hemolytic streptococci and Staphylococcus aureus and pathogenic viruses of upper respiratory tract infections, and can also enhance immunity, reduce inflammation and coagulation; Dandelion contains various healthy nutrients such as taraxacinol, taraxacin, choline, organic acid, inulin, etc., which have diuretic, laxative, anti-jaundice, and choleretic effects; Patchouli contains volatile oils and flavonoid compounds. Chicory is rich in oligofructose, inulin, dietary fiber, vitamin C and E, and has the effects of regulating the balance of intestinal flora, improving intestinal health, regulating blood sugar and blood lipids, slowing down cell aging and inflammatory response; Hawthorn is rich in organic acids, flavonoids and triterpenoid saponins, and has the effects of digesting food, removing accumulation, promoting qi and relieving blood stasis; Black plum mainly contains organic acid components, including citric acid, malic acid and ursolic acid, and has the effects of promoting fluid production and quenching thirst, astringing the lungs and relieving cough; The above Chinese medicine ingredients are combined and fermented with Aspergillus niger to prepare a composite enzyme preparation, which has enzymes with multiple functions, can effectively release the Chinese medicine ingredients and improve the utilization rate of Chinese medicine.

[0009] At the same time, the present application adopts an appropriate weight ratio to mix Aspergillus niger, basic nutrients and traditional Chinese medicine, and fully releases nutrients and traditional Chinese medicine active factors after fermentation. Moreover, the active ingredients of traditional Chinese medicine have a proper ratio, which synergistically exerts the effects of improving immunity and adjusting intestinal health.

[0010] Preferably, the weight ratio of the Aspergillus niger, the basic nutrients and the traditional Chinese medicine is 1:160-170:55-65.

[0011] Preferably, the basic nutrition comprises bran and peanut shells.

[0012] By adopting the above technical scheme, bran is used as energy feed to provide nutrition, and peanut shells are used as fiber feed to play a ventilation role. Under the fermentation of Aspergillus niger, rich secondary metabolites are obtained, mainly alkaloids, organic acids, polysaccharides, flavonoids, total phenols and a wide variety of enzymes; the effective ingredients of traditional Chinese medicine are increased, while the utilization rate of feed is improved and the palatability of feed is increased.

[0013] Preferably, the basic nutrition consists of a mixture of bran and peanut shells in a weight ratio of 8-10:4-6.

[0014] Preferably, the traditional Chinese medicine is composed of the following components in parts by weight: 25-30 parts of coptis root, 20-25 parts of honeysuckle, 15-20 parts of dandelion, 10-12 parts of patchouli, 5-8 parts of chicory, 5-8 parts of hawthorn, and 5-8 parts of ebony.

[0015] Preferably, the complex enzyme preparation comprises acid protease, pectinase, amylase, xylanase, cellulase and glucanase.

[0016] Preferably, in the composite enzyme preparation: the enzyme activity of the acid protease is 20000-40000 U / g, the enzyme activity of the pectinase is 7000-9000 U / g, the enzyme activity of the amylase is 1000-2000 U / g, the enzyme activity of the xylanase is 1000-2100 U / g, the enzyme activity of the cellulase is 1000-2000 U / g, and the enzyme activity of the glucanase is 600-800 U / g.

[0017] By adopting the above technical scheme of the present application, high-activity, multi-type enzymes can be obtained, and the release of effective ingredients of traditional Chinese medicine can be accelerated, thereby improving the utilization rate of traditional Chinese medicine, and having the effects of improving immunity and regulating intestinal health.

[0018] In a second aspect, the present application provides a method for preparing the above-mentioned composite enzyme preparation, which is characterized in that it specifically includes the following steps in sequence: preparation of Aspergillus niger spore suspension, initial solid fermentation culture, turning over and feeding, and drying after the fermentation culture is completed to obtain the composite enzyme preparation.

[0019] Preferably, the preparation method of the complex enzyme preparation specifically comprises the following steps in sequence: Preparation of Aspergillus niger spore suspension: Inoculate a single colony of Aspergillus niger on a slant medium and culture until the spores mature; then wash the mature spores with physiological saline, put them into a bottle, shake them on a shaker to break up the spores; filter the spore suspension, dilute the suspension with physiological saline, and adjust the final spore concentration to 0.5-1.5×10 7 / mL, and a spore suspension was prepared; Solid fermentation initial culture: the basic nutrients and the traditional Chinese medicine are placed in a grinder for crushing and sieving to obtain a powder culture medium, the Aspergillus niger spore suspension is added, and sterile water is added, and the mixture is stirred evenly to obtain a fermentation mixture, and then fermented and cultured in an environment of 30-35°C; wherein the weight ratio of the powder culture medium to the sterile water is 3-5:4; Turning over the koji and supplementing the feed: according to the percentage of the nutrient solution relative to the weight of the powder culture medium, supplementing 1.0-1.5% of the nutrient solution on the 2nd to 4th day of cultivation, supplementing 0.5-1.0% of the nutrient solution on the 5th to 7th day of cultivation; controlling the water content of the fermentation mixture to be 35%-50%; The composition of the nutrient solution is: 1.8-2.2% glucose, 26-30% defatted soybean powder, 3.5-4.5% peptone, 2-2.4% potassium dihydrogen phosphate, 0.5-0.7% magnesium sulfate, which are dissolved in potato juice and the pH value is adjusted to 5.8-6.3 with 4-6 mol / L NaOH solution; drying: after fermentation and culture for 6-8 days, the fermentation mixture is dried until the water content is controlled to be ≤10%, and crushed to obtain a composite enzyme preparation.

[0020] By adopting the above-mentioned technical scheme, the present application develops Aspergillus niger through aseptic inoculation and has strong fermentation activity. The Aspergillus niger seed liquid is added to a moist solid culture medium for solid fermentation culture, releasing a variety of active enzymes and active Chinese medicine ingredients. Turning over the koji and supplementing the feed promote the full progress of the fermentation process, and a composite enzyme preparation rich in a variety of enzymes and active Chinese medicine substances is obtained.

[0021] The preparation method of the composite enzyme preparation provided in the present application is simple to operate, has low energy consumption, is easy to control the fermentation process, has high production efficiency per unit volume, and can effectively improve the substrate.

[0022] In a third aspect, the present application provides the use of the above-mentioned complex enzyme preparation in the preparation of feed additives. By adopting the above technical scheme, a complex enzyme preparation with both medicinal and nutritional properties can be obtained. The complex enzyme preparation can be used as a feed additive in livestock breeding. It can supplement a variety of functional enzymes and free active Chinese medicine ingredients, which helps to improve feed quality and promote livestock health, which is in line with the development direction of sustainable green breeding.

[0023] In summary, the technical solution of this application has the following effects: The composite enzyme preparation provided in the present application has high enzyme activity and a wide enzyme system. It can effectively degrade multi-component anti-nutritional factors and improve feed utilization. At the same time, it can be used in feed as a poultry and livestock feed additive to enhance immunity and regulate intestinal health, which is in line with the development direction of sustainable green breeding.

[0024] The acidic protease activity in the compound enzyme preparation provided by the present application is as high as 20000-40000U / g. In the case of low protein, the effective amino acid supply should be more balanced; the protease eliminates anti-nutritional factors, especially anti-nutritional factors of plant protein sources, such as antigenic proteins, lectins, antitrypsin, etc. Therefore, the compound enzyme preparation provided by the present application can be well applied to low-protein feed as a feed additive.

[0025] The solid fermentation medium of the composite enzyme preparation of the present application is a mixture of Chinese medicine particles and basic nutrients. Through the fermentation of Aspergillus niger, the effective ingredients of Chinese medicine can be effectively released in large quantities, with the alkaloids increased by more than 1.59 times, the total acid increased by more than 1.65 times, the total flavonoids increased by more than 5.46 times, and the total phenols increased by more than 3.26 times. The utilization rate and palatability of Chinese medicine are improved, and it has both medicinal and nutritional properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is an appearance picture of the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger; wherein, the left tray contains the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger in Example 1, and the right tray contains the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger in Example 2. DETAILED DESCRIPTION

[0027] The present application is further described in detail below in conjunction with examples, comparative examples and performance testing. These examples should not be construed as limiting the scope of protection claimed in the present application. It should be 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. Unless otherwise specified, the raw materials used in the following examples can all be sourced from ordinary commercial sources. Example

[0028] Example 1 Example 1 provides a composite enzyme preparation of traditional Chinese medicine fermented by Aspergillus niger and a preparation method thereof.

[0029] The preparation method of the composite enzyme preparation in Example 1 specifically comprises the following steps in sequence: (1) Preparation of Aspergillus niger spore suspension: Inoculate 1g of Aspergillus niger colony into PDA slant medium and culture at 30℃ for 100h until spores mature; then wash the mature spores with saline, put them into a triangular flask containing glass beads, shake on a shaker for 2h to break up the spores. Filter the spore suspension with four layers of gauze, dilute the suspension with an appropriate amount of saline, count the spore concentration with a hemocytometer, and adjust the final spore concentration to 10 7 / mL, and 70mL of Aspergillus niger spore suspension was prepared.

[0030] (2) Solid fermentation initial culture: Take dry bran and dry peanut shell powder and mix them in a weight ratio of 9:5 to obtain basic nutrients; 27 g of coptis root, 22 g of honeysuckle, 17 g of dandelion, 11 g of patchouli, 6 g of chicory, 6 g of hawthorn, and 6 g of ebony are mixed to obtain Chinese medicine; 165 g of basic nutrients and 60 g of Chinese medicine are put into a grinder and ground until all of them pass through a 20-mesh sieve to obtain a powder as a powder culture medium; Add the Aspergillus niger spore suspension to the powder medium, add 90 g (225 g) of sterile water, stir evenly to obtain a fermentation mixture, cover with a cloth, and then ferment and culture under natural pH conditions and 32° C. (3) Turning over the koji and adding materials: According to the percentage of nutrient solution to the weight of powder culture medium, 1.25% nutrient solution is supplemented on the 3rd day of culture, and 0.75% nutrient solution is supplemented on the 5th day of culture; the koji is turned over each time when supplementing, and the water content of the fermentation mixture is tested after each turning over. When the water content is lower than 35%, sterile water is supplemented, and the water content is controlled not to exceed 55%; the water content of the fermentation mixture is controlled to be 35%-50%.

[0031] In this example, sterile water was added on the 3rd day until the water content of the fermentation mixture was 45%, and sterile water was added on the 5th day until the water content of the fermentation mixture was 45%.

[0032] The composition of the nutrient solution is: 2% glucose, 28% defatted soybean powder, 4% peptone, 2.2% potassium dihydrogen phosphate, 0.6% magnesium sulfate, which are dissolved in potato juice and the pH is adjusted to 6 with 5 mol / L NaOH solution.

[0033] (5) Drying: After the fermentation mixture is fermented and cultured at a constant temperature of 30°C for 7 days, the koji is covered with black spores. The fermentation mixture is evaporated at 45°C to a moisture content of ≤10%, and then crushed and preserved to obtain a composite enzyme preparation rich in acid protease, pectinase, amylase, xylanase, cellulase and glucanase.

[0034] Embodiment 2-5 Examples 2-5 respectively provide a composite enzyme preparation of a Chinese medicine fermented by Aspergillus niger and a preparation method thereof.

[0035] The difference between the above embodiment and embodiment 1 is that the weight ratios of Aspergillus niger, basic nutrients and traditional Chinese medicine are different, as shown in Table 1.

[0036] Table 1 Weight ratio of Aspergillus niger, basic nutrients and Chinese medicine in Examples 1-5 Example Weight ratio of Aspergillus niger, basic nutrients and Chinese medicine 1 The weight ratio of Aspergillus niger, basic nutrients and Chinese medicine is 1:165:60 2 The weight ratio of Aspergillus niger, basic nutrients and Chinese medicine is 1:160:65 3 The weight ratio of Aspergillus niger, basic nutrients and Chinese medicine is 1:170:55 4 The weight ratio of Aspergillus niger, basic nutrients and Chinese medicine is 1:150:70 5 The weight ratio of Aspergillus niger, basic nutrients and Chinese medicine is 1:180:50 The other process parameters in the above embodiment are the same as those in embodiment 1.

[0037] Embodiment 6-9 Examples 6-9 respectively provide a composite enzyme preparation of a Chinese medicine fermented by Aspergillus niger and a preparation method thereof.

[0038] The difference between the above embodiment and embodiment 1 is that the composition of the Chinese medicine is different, as shown below.

[0039] In Example 6: 25 g of Rhizoma Coptidis, 25 g of Flos Lonicerae, 15 g of Herba Taraxaci, 12 g of Herba Patchouli, 5 g of Cichorium, 8 g of Fructus Crataegi and 5 g of Prunus Mume were mixed to obtain a Chinese medicine.

[0040] In Example 7: 30 g of Rhizoma Coptidis, 20 g of Flos Lonicerae, 20 g of Herba Taraxaci, 10 g of Herba Patchouli, 8 g of Cichorium Indicum, 5 g of Crataegus, and 8 g of Prunus Mume were mixed to obtain a traditional Chinese medicine.

[0041] In Example 8: 22 g of Rhizoma Coptidis, 27 g of Flos Lonicerae, 12 g of Herba Taraxaci, 14 g of Herba Patchouli, 9 g of Cichorium Indicum, 4 g of Crataegus, and 9 g of Prunus Mume were mixed to obtain a traditional Chinese medicine.

[0042] In Example 9: 32 g of Coptis chinensis, 17 g of Honeysuckle, 22 g of Taraxacum mongolicum, 8 g of Patchouli, 4 g of Cichorium indicum, 9 g of Crataegus pinnatifida, and 4 g of Prunus mume were mixed to obtain a traditional Chinese medicine.

[0043] The other process parameters in the above embodiment are the same as those in embodiment 1.

[0044] Examples 10-13 Examples 10-13 respectively provide a composite enzyme preparation of a Chinese medicine fermented by Aspergillus niger and a preparation method thereof.

[0045] The differences between the above embodiment and embodiment 1 are as follows: the steps of turning over the koji and adding materials are different, as shown below.

[0046] In Example 10: according to the percentage of nutrient solution relative to the weight of powder culture medium, 1.0% nutrient solution was supplemented on the second day of culture to control the moisture content to 35%; 1.0% nutrient solution was supplemented on the sixth day of culture to control the moisture content to 50%.

[0047] In Example 11: according to the percentage of nutrient solution relative to the weight of powder culture medium, 1.5% nutrient solution was supplemented on the 4th day of culture to control the water content to 50%; 0.5% nutrient solution was supplemented on the 7th day of culture to control the water content to 35%.

[0048] In Example 12: according to the percentage of nutrient solution relative to the weight of powder culture medium, 0.75% nutrient solution was supplemented on the 3rd day of culture to control the moisture content to 30%; 1.25% nutrient solution was supplemented on the 5th day of culture to control the moisture content to 55%.

[0049] In Example 13: according to the percentage of nutrient solution relative to the weight of powder culture medium, 1.75% nutrient solution was supplemented on the 3rd day of culture to control the water content to 55%; 0.5% nutrient solution was supplemented on the 5th day of culture to control the water content to 30%.

[0050] The other process parameters in the above embodiment are the same as those in embodiment 1.

[0051] Comparative Example Comparative Examples 1-4 Comparative Examples 1-4 respectively provide a composite enzyme preparation of a Chinese medicine fermented by Aspergillus niger and a preparation method thereof.

[0052] The difference between the above comparative example and Example 1 is that the steps of turning over the koji and supplementing the material are different, as shown below.

[0053] In Comparative Example 1, the weight ratio of Aspergillus niger, basic nutrients and traditional Chinese medicine is 1:140:80.

[0054] In Comparative Example 2, the weight ratio of Aspergillus niger, basic nutrients and traditional Chinese medicine is 1:200:40.

[0055] In comparative example 3: in the Chinese medicine composition, an equal amount of Scutellaria baicalensis is used to replace Coptis chinensis.

[0056] In comparative example 4: 18 g of coptis root, 35 g of honeysuckle, 10 g of dandelion, 16 g of patchouli, 2 g of chicory, 10 g of hawthorn and 2 g of ebony are mixed to obtain a traditional Chinese medicine.

[0057] The other process parameters in the above comparative example are the same as those in Example 1.

[0058] Performance testing like Figure 1 The figure shows the appearance of the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger; wherein, the left tray contains the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger in Example 1, and the right tray contains the complex enzyme preparation obtained after the Chinese medicine is fermented by Aspergillus niger in Example 2.

[0059] (1) Enzyme activity: The composite enzyme preparations prepared in the examples and comparative examples were subjected to index determination. The enzyme activity determination method of each enzyme was as follows: the enzyme activity determination of acid protease was referred to GB / T 23527-2009, the enzyme activity determination of pectinase was referred to QB / T 4482-2013, the enzyme activity determination of amylase was referred to QB / T1803-1993, the enzyme activity determination of xylanase was referred to GB / T 23874-2009, the enzyme activity determination of cellulase was referred to NYT 912-2020, and the enzyme activity determination of glucanase was referred to NYT 911-2020.

[0060] The specific enzyme activity results are shown in Table 2.

[0061] Table 2 Enzyme activity of composite enzyme preparations of Chinese medicine fermented by Aspergillus niger in Examples and Comparative Examples As can be seen from Table 2, the compound enzyme preparation of Aspergillus niger fermented Chinese medicine has many types of enzymes and high enzyme activity, especially the acid protease with the highest enzyme activity, followed by pectinase. Acid protease has complex physical and chemical properties. It can catalyze and hydrolyze protein under acidic conditions. As an additive, it can promote the growth and development of young animals and reduce the stress caused by weaning. It is widely used in animal husbandry and has broad development prospects.

[0062] (2) Active substances of traditional Chinese medicine: The active substances of traditional Chinese medicine were determined by taking the mixture before fermentation as the control. The determination of alkaloids was based on the high performance liquid chromatography method, the determination of organic acids was based on the total acid titration method, the determination of total flavonoids was based on the aluminum nitrate colorimetric method, and the determination of total phenols was based on the Folin-phenol colorimetric method.

[0063] The specific results are shown in Table 3.

[0064] Table 3 Content of active substances in Chinese medicine fermented by Aspergillus niger in the embodiment As can be seen from Table 3, Aspergillus niger can use the various enzymes secreted by itself to degrade or transform the active ingredients of traditional Chinese medicine, so that the effective ingredients of traditional Chinese medicine are fully released, the alkaloids are increased by more than 1.59 times, the total acid is increased by more than 1.65 times, the total flavonoids are increased by more than 5.46 times, and the total phenols are increased by more than 3.26 times. Aspergillus niger ferments traditional Chinese medicine and can transform its macromolecular substances into small molecular substances that can be directly absorbed by the intestines, which can be used to prevent and treat various diseases, and can also improve the flavor of traditional Chinese medicine and improve its palatability.

[0065] Application Example 1 The compound enzyme preparation prepared in Example 1 was added to ordinary pig feed to carry out a feeding effect test. 30 piglets aged 1-5 weeks in the farm were used as test subjects, and three groups of 10 pigs were selected, including 1 blank control group and 2 comparative test groups. The piglets in the blank control group were raised normally, and the compound enzyme preparation of the present application was not added during the weaning period. The compound enzyme preparation of the present application was added to the piglets in the two test groups during the weaning period, and the feeding time was one week. This application has statistically analyzed the weight gain and feed-to-meat ratio improvement of the above test groups, and the specific situation is shown in Table 4 below.

[0066] Table 4: Weight gain and feed-to-meat ratio improvement of the composite enzyme preparation prepared in Example 1 in pig feed Group Weight gain Feed to meat ratio Improvement in feed-to-meat ratio (%) Blank control group 60％ 1.31 / Example 1 - Comparative Test 1 85％ 1.23 6.11 Example 1 - Comparative Test 2 90％ 1.21 7.63 In Table 4, the feed-to-meat ratio is the ratio of the weight of feed input to the weight gain of the pigs, and the improvement of the feed-to-meat ratio = (feed-to-meat ratio of the blank control - feed-to-meat ratio of the comparative test) / feed-to-meat ratio of the blank control × 100%.

[0067] Based on the above results, it can be seen that adding a composite enzyme preparation of Aspergillus niger fermented Chinese medicine to piglet feed can supplement the endogenous enzymes in the piglet to a certain extent, decompose the high-molecular protein in the feed into low-molecular amino acids, and help the piglet digest and absorb nutrients. The active ingredients of Chinese medicine can reduce the stimulation of feed to the piglet's digestive system and reduce stress reactions. The piglet can be unaffected by weaning in the early development stage, ensuring the timely supply of nutrients in the growth stage. Therefore, the composite enzyme preparation provided by the present application can be used as a feed additive in feed, effectively improving the immunity of the piglet and regulating intestinal health.

[0068] 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 of Aspergillus niger fermented Chinese medicine, characterized in that: The composite enzyme preparation is prepared by fermenting a mixture of basic nutrients and traditional Chinese medicine using Aspergillus niger; The weight ratio of the Aspergillus niger, the basic nutrients and the traditional Chinese medicine is 1:150-180:50-70; The traditional Chinese medicine consists of the following components in parts by weight: 22-32 parts of coptis root, 17-27 parts of honeysuckle, 12-22 parts of dandelion, 8-14 parts of patchouli, 4-9 parts of chicory, 4-9 parts of hawthorn and 4-9 parts of ebony.

2. The complex enzyme preparation according to claim 1, characterized in that The weight ratio of the Aspergillus niger, the basic nutrients and the traditional Chinese medicine is 1:160-170:55-65.

3. The complex enzyme preparation according to claim 1, characterized in that The basic nutrients include bran and peanut shells.

4. The complex enzyme preparation according to claim 3, characterized in that The basic nutrition consists of a mixture of bran and peanut shells in a weight ratio of 8-10:4-6.

5. The complex enzyme preparation according to claim 1, characterized in that The traditional Chinese medicine consists of the following components in parts by weight: 25-30 parts of coptis root, 20-25 parts of honeysuckle, 15-20 parts of dandelion, 10-12 parts of patchouli, 5-8 parts of chicory, 5-8 parts of hawthorn and 5-8 parts of ebony.

6. The complex enzyme preparation according to claim 1, characterized in that The complex enzyme preparation comprises acid protease, pectinase, amylase, xylanase, cellulase and glucanase.

7. The complex enzyme preparation according to claim 6, characterized in that In the composite enzyme preparation: the enzyme activity of the acid protease is 20000-40000 U / g, the enzyme activity of the pectinase is 7000-9000 U / g, the enzyme activity of the amylase is 1000-2000 U / g, the enzyme activity of the xylanase is 1000-2100 U / g, the enzyme activity of the cellulase is 1000-2000 U / g, and the enzyme activity of the glucanase is 600-800 U / g.

8. The method for preparing the complex enzyme preparation according to any one of claims 1 to 7, characterized in that: Specifically, the following steps are sequentially performed: preparation of Aspergillus niger spore suspension, initial solid fermentation culture, turning over and feeding, and drying after the fermentation culture is completed to obtain the composite enzyme preparation.

9. The method for preparing the complex enzyme preparation according to claim 8, characterized in that: Specifically, the following steps are performed in sequence: Preparation of Aspergillus niger spore suspension: Inoculate a single colony of Aspergillus niger on a slant medium and culture until the spores mature; then wash the mature spores with physiological saline, put them into a bottle, shake them on a shaker to break up the spores; filter the spore suspension, dilute the suspension with physiological saline, and adjust the final spore concentration to 0.5-1.5×10 7 / mL, and a spore suspension was prepared; Solid fermentation initial culture: the basic nutrients and the traditional Chinese medicine are placed in a grinder for crushing and sieving to obtain a powder culture medium, the Aspergillus niger spore suspension is added, and sterile water is added, and the mixture is stirred evenly to obtain a fermentation mixture, and then fermented and cultured in an environment of 30-35°C; wherein the weight ratio of the powder culture medium to the sterile water is 3-5:4; Turning over the koji and supplementing the feed: according to the percentage of the nutrient solution relative to the weight of the powder culture medium, supplementing 1.0-1.5% of the nutrient solution on the 2nd to 4th day of cultivation, supplementing 0.5-1.0% of the nutrient solution on the 5th to 7th day of cultivation; controlling the water content of the fermentation mixture to be 35%-50%; The composition of the nutrient solution is: 1.8-2.2% glucose, 26-30% defatted soy flour, 3.5-4.5% peptone, 2-2.4% potassium dihydrogen phosphate, 0.5-0.7% magnesium sulfate, which are dissolved in potato juice and adjusted to pH 5.8-6.3 with 4-6 mol / L NaOH solution; Drying: After 6-8 days of fermentation, the fermentation mixture is dried until the moisture content is controlled to ≤10%, and then crushed to obtain the complex enzyme preparation.

10. Use of the complex enzyme preparation according to any one of claims 1 to 7 in the preparation of feed additives.

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