CN117904078A - Complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine and preparation method and application thereof - Google Patents

Abstract

The application relates to the technical field of traditional Chinese medicine fermentation, in particular to a compound enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine, and a preparation method and application thereof. The complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine is obtained by using a mixture of aspergillus oryzae fermented basic nutrition and traditional Chinese medicine. The Chinese medicinal materials comprise folium Artemisiae Argyi, caulis Zanthoxyli, herba Taraxaci, flos Lonicerae, pericarpium Citri Tangerinae and Glycyrrhrizae radix. The preparation method comprises the steps of preparing Aspergillus oryzae seed liquid, crushing and mixing the traditional Chinese medicine and basic nutrition as a culture medium, inoculating Aspergillus oryzae into a solid culture medium, fermenting, and finally fermenting to obtain the compound enzyme preparation. The complex enzyme preparation prepared by the application has the advantages of wide enzyme system, high activity of neutral protease, rich bioactive components such as polysaccharide, flavone, glycoside and the like, capability of effectively improving the immunity of livestock, simple preparation method, low cost for production and subsequent environment-friendly treatment, and the complex enzyme preparation has two properties of medicine and nutrition.

Description

Complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine and preparation method and application thereof

Technical Field

The application relates to the technical field of fermentation, in particular to a compound enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine, a preparation method and application thereof.

Background

Aspergillus oryzae (Aspergillus oryzae), belonging to the phylum Deuteromycotina, class Cellularomycetes, family Conidiosporaceae, is a common non-toxic Aspergillus fungus. Aspergillus oryzae is a strain producing complex enzymes, mainly producing protease, amylase, saccharifying enzyme, cellulase, phytase, etc. Aspergillus oryzae contains rich glycoside hydrolase and can be used for efficiently producing protease by using starch or cellulose and other raw materials. The proteases produced by Aspergillus oryzae are of a wide variety, and the Aspergillus oryzae proteases currently detected include 9 acid proteases, 3 neutral proteases and 6 alkaline proteases. Aspergillus oryzae protease has wide pH tolerance range, high temperature resistance and good stability.

Neutral protease is an important protease secreted by Aspergillus oryzae, and has an optimal reaction temperature of 55deg.C and an optimal reaction pH of 7.0-7.2. The neutral protease can decompose macromolecular proteins into small peptides or amino acids, and the neutral protease is added into the feed, so that the utilization rate of the proteins is improved, and the feeding cost is reduced.

The existing processing technology of traditional Chinese medicines can be divided into four types, namely modification, water preparation, fire preparation and water-fire co-preparation, and the preparation methods of the 4 traditional Chinese medicines are all long, but all the effective components of the traditional Chinese medicines are changed and the drug effect is reduced. The enzymatic traditional Chinese medicine, namely the traditional Chinese medicine prepared by adopting a fermentation method, has no damage to the active ingredients of the traditional Chinese medicine, increases the original efficacy by times, and also breaks down some ingredients into small molecule traditional Chinese medicine ingredients. The enzymatic micromolecular Chinese medicine not only can be completely absorbed, but also has the function of treating and repairing genes.

As a substitute for feed antibiotics, the common veterinary traditional Chinese medicine has the efficacy of improving the immunity of livestock and poultry, has the advantages of no toxic or side effect, no medicine residue and the like, but has the defects of poor palatability, slow effect, low utilization rate and the like in the practical application process.

Disclosure of Invention

In view of the defects of poor palatability, low utilization rate, slow effect in improving the immunity of livestock and poultry and the like of the common veterinary traditional Chinese medicine in the practical application process, the application uses aspergillus oryzae to effectively ferment the traditional Chinese medicine so as to overcome the defects.

In a first aspect, the application provides a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine, and adopts the following technical scheme.

A complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine, which is obtained by using a mixture of aspergillus oryzae fermented basic nutrition and traditional Chinese medicine; the Chinese medicine is a composition of mugwort leaf, pricklyash peel stem and leaf, dandelion, honeysuckle, dried orange peel and liquorice.

By adopting the technical scheme, the aspergillus oryzae ferments the traditional Chinese medicine to generate a plurality of high-activity enzymes, thereby accelerating the release of the effective components of the traditional Chinese medicine, improving the utilization rate of the traditional Chinese medicine and improving the taste.

As an improvement of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the basic nutrients comprise bean pulp and peanut shells.

By adopting the technical scheme, the soybean meal provides rich proteins and enzymes, the peanut shell contains 60% of crude fibers, under the fermentation of aspergillus oryzae, nutrient substances such as peptide chains, amino acids, glucose and the like are released, and rich enzymes are produced, so that the release of active ingredients of the traditional Chinese medicine can be accelerated, the utilization rate of the traditional Chinese medicine is improved, and the taste is improved.

As an improvement of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the complex enzyme preparation comprises neutral protease, xylanase, cellulase, pectinase, amylase and saccharifying enzyme; the enzyme activity of the neutral protease is 60000-70000U/g, the enzyme activity of the xylanase is 8000-10000U/g, the enzyme activity of the cellulase is 9000-10000U/g, the enzyme activity of the pectase is 1900-3200U/g, the enzyme activity of the amylase is 900-920U/g, and the enzyme activity of the saccharifying enzyme is 900-990U/g.

By adopting the technical scheme, the release of the active ingredients of the traditional Chinese medicine can be accelerated by high-activity and various enzymes, the utilization rate of the traditional Chinese medicine is improved, the taste is improved, and the effects of improving the immunity and the growth speed are achieved.

In a second aspect, the application also provides a preparation method of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, and the following technical scheme is adopted.

A preparation method of a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine comprises the steps of obtaining the complex enzyme preparation by using aspergillus oryzae through solid state fermentation of basic nutrients and traditional Chinese medicine; the Chinese medicine is a composition of mugwort leaf, pricklyash peel stem and leaf, dandelion, honeysuckle, dried orange peel and liquorice.

By adopting the technical scheme: the mugwort leaf contains various active ingredients such as olein, alkene, alcohol and ketone which can inhibit bacteria, stop bleeding, relieve itching, relieve asthma and protect liver and gall; the stems and leaves of the wild pepper contain volatile oil, alkaloid, lignan, flavone, amide compounds, coumarin, fatty acid and other bioactive substances, and have the effects of killing insects, dispelling wind and cold, activating blood and relieving pain; the dandelion is rich in folic acid and copper, and has the effects of clearing heat and detoxicating, detumescence and resolving hard mass, and inducing diuresis and relieving diarrhea; the honeysuckle contains honeysuckle extract, rosmarinic acid, kaempferic acid and the like, and has the functions of resisting oxidization, inflammation and bacteria and enhancing immunity; the pericarpium Citri Tangerinae contains hesperidin, rutin, naringin, neohesperidin, nobiletin, and tangerines, and has effects of promoting secretion of digestive juice, removing intestinal qi, dilating coronary artery, and promoting bile flow; glycyrrhrizae radix mainly contains flavonoids, coumarin, triterpene saponin and triterpene aglycone, and has effects of treating spleen and stomach weakness, listlessness debilitation, palpitation, short breath, cough with excessive phlegm, carbuncle, skin sore, and spasm pain of stomach and abdomen, limbs; the composite enzyme preparation is prepared by combining the traditional Chinese medicine components and fermenting aspergillus oryzae, has a plurality of high-activity enzymes, can accelerate the release of the effective components of the traditional Chinese medicine, improves the utilization rate of the traditional Chinese medicine and improves the taste.

As an improvement of the preparation method of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the mass ratio of the aspergillus oryzae to the basic nutrition to the traditional Chinese medicine is 1: (140-180): (30-50).

By adopting the technical scheme, proper material proportion is adopted, and nutrients and traditional Chinese medicine active factors are fully released through fermentation.

As an improvement of the preparation method of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the mass content of each component in the traditional Chinese medicine is as follows: 35-40% of mugwort leaf, 20-25% of pepper stem and leaf, 15-20% of dandelion, 10-12% of honeysuckle, 4-8% of dried orange peel and 4-7% of liquorice.

By adopting the technical scheme, the traditional Chinese medicine has proper proportion of active ingredients, and the effects of improving immunity, improving taste and promoting healthy growth of animals are synergistically exerted.

As an improvement of the preparation method of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the basic nutrition is a composition of soybean meal and peanut shell, and the soybean meal is as follows: the mass ratio of peanut shells is = (9-11) = (5-7).

By adopting the technical scheme, under the fermentation of aspergillus oryzae, various nutrients and various active enzymes are released, and the release of active ingredients of the traditional Chinese medicine is promoted.

As an improvement of the preparation method of the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine, the preparation method comprises the following steps: preparing aspergillus oryzae seed liquid: inoculating Aspergillus oryzae into sterile water for culturing to obtain Aspergillus oryzae seed liquid;

solid fermentation primary culture: preparing the basic nutrients and the traditional Chinese medicines into powder, taking the powder as a culture medium, adding the aspergillus oryzae seed liquid, uniformly mixing to obtain a fermentation mixture, and carrying out fermentation culture;

Turning and feeding: supplementing a carbon source and a nitrogen source in the fermentation culture process, turning over, and controlling the water content of the fermentation mixture; and after fermentation culture is completed, the compound enzyme preparation is prepared.

By adopting the technical scheme, the aspergillus oryzae asepsis inoculation development has stronger fermentation activity, the aspergillus oryzae seed liquid is added into a wet solid culture medium for solid fermentation culture, various active enzymes and active traditional Chinese medicine components are released, the fermentation process is fully carried out by turning and feeding, and the compound enzyme preparation rich in various active enzymes and active traditional Chinese medicine components is obtained.

As an improvement of the preparation method of the aspergillus oryzae fermented traditional Chinese medicine compound enzyme preparation, in the preparation of the aspergillus oryzae seed liquid, the aspergillus oryzae is inoculated into sterile water and then is cultured for 4-8 hours in a constant temperature incubator at 30-35 ℃ to obtain the aspergillus oryzae seed liquid.

During the primary culture of the solid fermentation, the basic nutrition and the traditional Chinese medicine are put into a pulverizer for pulverizing and sieving to obtain powder, the powder is taken as a culture medium, the aspergillus oryzae seed liquid is added, sterile water is supplemented, the mixture is stirred uniformly to obtain a fermentation mixture, and then the fermentation mixture is fermented and cultured in an environment of 30-35 ℃; in the fermentation mixture which is preliminarily prepared, the mass of the aspergillus oryzae is 0.3-0.7% of the mass of the culture medium, and the mass ratio of the culture medium to the total amount of sterile water is (4-6): 3.

Among the turning and feeding: the carbon source is glucose, and the nitrogen source is ammonium sulfate; according to the percentages of carbon sources and nitrogen sources relative to the mass of the culture medium, 0.4-0.6% of glucose and 0.4-0.6% of ammonium sulfate are supplemented on days 2-4, 0.4-0.6% of glucose and 0.4-0.6% of ammonium sulfate are supplemented on days 5-7, and 0.1-0.3% of glucose and 0.1-0.3% of ammonium sulfate are supplemented on days 8-10; the water content of the fermentation mixture is controlled to be 30% -55%.

After the fermentation mixture is fermented and cultured for 11-13 days, the water content of the fermentation mixture is controlled to be less than or equal to 10%, and the fermentation mixture is crushed to obtain the compound enzyme preparation.

By adopting the technical scheme, the preparation method is simple and energy-saving, has no wastewater pollution, has high production efficiency per unit volume, and can lead the substrate to be changed benign.

In a third aspect, the present application provides an application of the complex enzyme preparation, and adopts the following technical scheme.

The application of the complex enzyme preparation as a feed additive.

By adopting the technical scheme, the complex enzyme preparation is used as a feed additive, can be applied to livestock raising, can supplement various functional enzymes and free active traditional Chinese medicine components, is beneficial to improving the immunity and health condition of livestock, and promotes the healthy growth of livestock.

In conclusion, the aspergillus oryzae fermented traditional Chinese medicine compound enzyme preparation and the preparation method and application thereof have the following beneficial effects:

1) The complex enzyme preparation provided by the application has high enzyme activity and wide enzyme system, can effectively degrade macromolecular proteins and cellulose, improves the utilization rate of feed, can effectively improve the health condition of livestock and poultry by being used as a feed additive in the feed, effectively reduces the feed conversion ratio, and has good application prospect.

2) The enzyme activity of the neutral protease of the compound enzyme preparation provided by the application is up to 60000-70000U/g, and the protease is added into typical bean pulp type daily ration, so that the low protein level of the daily ration can be reduced by 2-3 percentage points according to NRC recommended standard. Protease is added into the mixed meal type daily ration, and the low protein level of the daily ration can be reduced by 3-4 percent. Protease improves the protein digestibility of the whole ration, which is the most important function of improving animal productivity and maintaining the productivity of low-protein ration.

3) The solid fermentation medium of the compound enzyme preparation is a mixture of traditional Chinese medicine and basic nutrients, the traditional Chinese medicine can be effectively enzymatically decomposed through the fermentation of aspergillus oryzae, the release of the effective components of the traditional Chinese medicine is accelerated, the utilization rate of the traditional Chinese medicine is improved, the taste is improved, the compound enzyme preparation has two properties of medicine and nutrition, and finally the effects of improving the immunity and regulating the health are achieved.

Drawings

The left tray in fig. 1 contains the complex enzyme preparation obtained after the aspergillus oryzae fermentation of the traditional Chinese medicine in example 1, and the right tray contains the complex enzyme preparation obtained after the aspergillus oryzae fermentation of the traditional Chinese medicine in example 2.

Detailed Description

The advantages and features of the present application will become more apparent from the following description when taken in conjunction with the accompanying drawings and detailed description. The specific description is: the following examples, in which no specific conditions are noted, are conducted under conventional conditions or conditions recommended by the manufacturer, and the raw materials used in the following examples are commercially available from ordinary sources except for the specific descriptions.

Aspergillus oryzae used in the examples below was obtained from yishui Jinrun Biotechnology Inc.

Example 1

A complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine is prepared by using aspergillus oryzae through a solid state fermentation mode, and the preparation method of the complex enzyme preparation specifically comprises the following steps:

(1) Preparing aspergillus oryzae seed liquid: 1g of Aspergillus oryzae powder is inoculated into 70mL of sterile water, and the mixture is cultured for 4 hours in a constant temperature incubator at 30 ℃ to obtain Aspergillus oryzae seed liquid.

(2) Solid fermentation primary culture: 110g of dried bean pulp, 50g of dried peanut shell powder and 40g of dried traditional Chinese medicine (the traditional Chinese medicine comprises 14g of mugwort leaf powder, 10g of Chinese prickly ash stem and leaf powder, 8g of dandelion, 4.8g of honeysuckle, 1.6g of dried orange peel and 1.6g of liquorice) are put into a pulverizer to be pulverized into powder with 20 meshes, the powder obtained after pulverizing and sieving is put into a tray, the powder is taken as a culture medium, aspergillus oryzae seed liquid prepared in the step (1) is added, 50mL of sterile water is supplemented, the mixture is stirred uniformly, a fermentation mixture is obtained, drawer cloth is covered, and then fermentation culture is carried out in a constant temperature environment at 32 ℃ under the natural pH condition. In the initial fermentation mixture, the inoculum size of the aspergillus oryzae is 0.5 percent, namely the mass of aspergillus oryzae powder/the mass of the culture medium is 1 g/the (110+50+40) g=0.5 percent, and the initial feed water mass ratio of the fermentation mixture is (110+50+40) g (70+50) g=5:3.

(3) Turning and feeding: during the fermentation culture of the fermentation mixture in a constant temperature environment at 32 ℃, 0.8g of glucose and 0.8g of ammonium sulfate are supplemented on the 3 rd day, 0.8g of glucose and 0.8g of ammonium sulfate are supplemented on the 6 th day, and 0.4g of glucose and 0.4g of ammonium sulfate are supplemented on the 9 th day. Turning the yeast during each feeding, detecting the water content of the fermentation mixture after each turning, feeding sterile water when the water content is lower than 30%, and controlling the water content to be not more than 55%. In this example, the water content of the fermentation mixture was 40% by supplementing the sterile water to the fermentation mixture on day 3, 45% by supplementing the sterile water to the fermentation mixture on day 6, and 40% by supplementing the sterile water to the fermentation mixture on day 9.

(4) And (3) drying: fermenting and culturing the fermentation mixture in a constant temperature environment at 32 ℃ for 11 days, wherein the yeast grows up to white hypha, evaporating the fermentation mixture at 45 ℃ until the water content is less than or equal to 10%, and crushing and preserving to obtain the compound enzyme preparation which is rich in neutral protease, xylanase, cellulase, pectase, amylase and diastase.

Example 2

A complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine is prepared by using aspergillus oryzae through a solid state fermentation mode, and the preparation method of the complex enzyme preparation specifically comprises the following steps:

(1) Preparing aspergillus oryzae seed liquid: 1g of Aspergillus oryzae powder is inoculated into 70mL of sterile water, and the mixture is cultured for 8 hours in a constant temperature incubator at 35 ℃ to obtain Aspergillus oryzae seed liquid.

(2) Solid fermentation primary culture: 90g of dried bean pulp, 70g of dried peanut shell powder and 40g of dried traditional Chinese medicine (the traditional Chinese medicine comprises 16g of mugwort leaf powder, 8g of Chinese prickly ash stem and leaf powder, 6g of dandelion, 4g of honeysuckle, 3.2g of dried orange peel and 2.8g of liquorice) are put into a pulverizer to be pulverized into powder with 20 meshes, the powder obtained after pulverizing and sieving is put into a tray, the powder is taken as a culture medium, aspergillus oryzae seed liquid prepared in the step (1) is added, 50mL of sterile water is supplemented, the mixture is stirred uniformly, a fermentation mixture is obtained, drawer cloth is covered, and then fermentation culture is carried out in a constant temperature environment at 35 ℃ under the natural pH condition. In the initial fermentation mixture, the inoculum size of the aspergillus oryzae is 0.5 percent, namely the mass of aspergillus oryzae powder/the mass of the culture medium is 1 g/the (90+70+40) g=0.5 percent, and the initial feed water mass ratio of the fermentation mixture is (90+70+40) g (70+50) g=5:3.

(3) Turning and feeding: during the fermentation culture of the fermentation mixture in a constant temperature environment at 32 ℃, 1.2g of glucose and 1.2g of ammonium sulfate are supplemented on the 3 rd day, 1.2g of glucose and 1.2g of ammonium sulfate are supplemented on the 6 th day, and 0.6g of glucose and 0.6g of ammonium sulfate are supplemented on the 9 th day. Turning the yeast during each feeding, detecting the water content of the fermentation mixture after each turning, feeding sterile water when the water content is lower than 30%, and controlling the water content to be not more than 55%. In this example, the water content of the fermentation mixture was 40% by supplementing the sterile water to the fermentation mixture on day 3, 45% by supplementing the sterile water to the fermentation mixture on day 6, and 40% by supplementing the sterile water to the fermentation mixture on day 9.

(4) And (3) drying: fermenting and culturing the fermentation mixture in a constant temperature environment at 32 ℃ for 11 days, wherein the yeast grows up to white hypha, evaporating the fermentation mixture at 45 ℃ until the water content is less than or equal to 10%, and crushing and preserving to obtain the compound enzyme preparation which is rich in neutral protease, xylanase, cellulase, pectase, amylase and diastase.

Example detection

As shown in FIG. 1, the left tray contains the complex enzyme preparation obtained after the Aspergillus oryzae fermentation of the Chinese medicine in example 1, and the right tray contains the complex enzyme preparation obtained after the Aspergillus oryzae fermentation of the Chinese medicine in example 2. The complex enzyme preparations prepared in example 1 and example 2 were subjected to index measurement, and the enzyme activities of the respective enzymes were measured by: the measurement of the enzyme activity of neutral protease refers to GB/T23527-2009, the measurement of the enzyme activity of xylanase refers to GB/T23874-2009, the measurement of the enzyme activity of cellulase refers to NYT 912-2020, the measurement of the enzyme activity of pectase refers to QB/T4482-2013, the measurement of the enzyme activity of amylase refers to QB/T1803-1993, and the measurement of the enzyme activity of saccharifying enzyme refers to QB/T1803-1993. Specific enzyme activities are shown in Table 1.

TABLE 1 enzyme Activity of Complex enzyme preparation of Aspergillus oryzae fermented traditional Chinese medicine

As can be seen from Table 1, the complex enzyme preparation of the traditional Chinese medicine fermented by Aspergillus oryzae has a plurality of enzyme types and high enzyme activity, and especially the enzyme activity of neutral protease is the highest, and xylanase and cellulase are the next. The complex enzyme preparation of Aspergillus oryzae fermented traditional Chinese medicine is also rich in bioactive components such as polysaccharide, flavone, glycoside, etc.

Common daily ration supplements some amino acids by adding amino acid additives, but common amino acid additives are lysine additives and methionine additives, and lack of supplement of limited amino acids such as valine, isoleucine and the like. The neutral protease of the complex enzyme preparation of the embodiment 1-2 has the enzyme activity as high as 60000-70000U/g, can improve the protein digestion rate of the whole ration, can supplement the supply of valine, isoleucine and other limiting amino acids, and ensures that the effective amino acid supply tends to be balanced under the condition of low-protein feed. The protease also eliminates antinutritional factors, especially those of vegetable protein origin, such as antigenic proteins, lectins, antitrypsin, etc. Therefore, the complex enzyme preparation provided by the application can be used as a feed additive to be applied to low-protein feed, and the immunity and the growth speed of the culture are improved.

Application example 1

The complex enzyme preparation prepared in example 1 was added to a normal pig feed. 50 pigs with the same growth condition in a farm are taken as test subjects and randomly divided into 5 groups, and 10 repeats of each group comprise 1 blank control group and 4 comparison test groups. The compound enzyme preparation of the application is not added into the pig diets of the blank control group, and the compound enzyme preparation prepared in the example 1 is added into the pig diets of 4 test groups (the addition amount is 400-1600g/t, namely 400-1600g of the compound enzyme preparation is added into each ton of the diets). The live pigs eat and drink water freely, immunity and insect expelling are carried out conventionally, and the test period is 40 days. The application counts the ratio of the ingredients to the meat of the test, and the specific conditions are shown in the following table 2:

table 2 feed to meat ratio improvement

In table 2, the feed/meat ratio is the ratio of the weight of the feed charged to the weight of the pig growing, and the feed/meat ratio improvement condition= (feed/meat ratio of the blank control-feed ratio of the comparison test)/(feed/meat ratio of the blank control x 100%).

Based on the results, the complex enzyme preparation of the embodiment 1 of the application can supplement the deficiency of endogenous enzyme, activate the secretion and activity of the endogenous enzyme, promote the degradation of coarse fodder, obviously reduce the feed conversion ratio of live pig cultivation, improve the feed conversion ratio by more than 10%, and reduce the feed conversion ratio along with the increase of the addition in the range of 400-1600g/t, and the feed conversion ratio tends to be stable after the addition reaches 1200g/t, so that the complex enzyme preparation improves the healthy growth condition of live pigs. Therefore, the complex enzyme preparation provided by the application can be used as a feed additive for degrading feed, and the economic benefit of feeding is effectively improved.

Application example test since the concentration of cytokines (IL-2, IL-6, TNF-alpha) in pig serum can reflect the level of cellular immunity of pigs, generally the higher the concentration of the factors is, the higher the immunity of pigs is, so the test pigs in application example 1 are fasted for 12 hours, 10mL of venous blood is taken, and after standing for 2 hours, serum is separated out, and centrifuged to prepare serum samples, and the ELISA method is used for measurement, the operation steps are carried out according to the specification of the kit, and the concentration of the cytokines (IL-2, IL-6, TNF-alpha) in pig serum is as shown in the following table 3:

TABLE 3 Effect of Complex enzyme preparation on pig cellular immunity

As can be seen from Table 3, the concentrations of cytokines (IL-2, IL-6, TNF-alpha) in the pig serum of test group 1, test group 2, test group 3 and test group 4 are higher than those of the control group, which indicates that the complex enzyme preparation of the application has the effect of improving the cellular immunity of pigs.

Some traditional Chinese medicines for pigs directly influence the efficacy exertion and the utilization rate of the traditional Chinese medicines after entering the alimentary canal because the effective components such as immune polysaccharide, total flavone and the like are wrapped by hemicellulose, cellulose and lignin. The compound enzyme preparation prepared by the application has an enzymatic function on traditional Chinese medicines, efficiently decomposes hemicellulose, cellulose and part of lignin, exposes effective components of the traditional Chinese medicines, and converts macromolecules into micromolecules and polysaccharides into active chemical components through enzyme conversion, so that after eating, the compound enzyme preparation can achieve a good effect of improving the immunity of pig organisms.

The above are merely some embodiments of the present application, the protective scope of the present application is not limited to the above embodiments, and it should be obvious to those skilled in the art that modifications and extensions according to the above embodiments are also within the protective scope of the present application without departing from the gist of the present application.

Claims (10)

1. A complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine, which is characterized in that the complex enzyme preparation is obtained by using a mixture of aspergillus oryzae fermented basic nutrition and traditional Chinese medicine; the Chinese medicine is a composition of mugwort leaf, pricklyash peel stem and leaf, dandelion, honeysuckle, dried orange peel and liquorice.

2. The complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to claim 1, wherein the basic nutrients comprise soybean meal and peanut shell.

3. The complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to claim 1, wherein the complex enzyme preparation comprises neutral protease, xylanase, cellulase, pectinase, amylase and saccharifying enzyme; the enzyme activity of the neutral protease is 60000-70000U/g, the enzyme activity of the xylanase is 8000-10000U/g, the enzyme activity of the cellulase is 9000-10000U/g, the enzyme activity of the pectase is 1900-3200U/g, the enzyme activity of the amylase is 900-920U/g, and the enzyme activity of the saccharifying enzyme is 900-990U/g.

4. A preparation method of a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine is characterized in that the complex enzyme preparation is obtained by using aspergillus oryzae through solid state fermentation of basic nutrients and traditional Chinese medicine; the Chinese medicine is a composition of mugwort leaf, pricklyash peel stem and leaf, dandelion, honeysuckle, dried orange peel and liquorice.

5. The method for preparing the complex enzyme preparation of the aspergillus oryzae fermented traditional Chinese medicine according to claim 4, wherein the mass ratio of the aspergillus oryzae to the basic nutrient to the traditional Chinese medicine is 1: (140-180): (30-50).

6. The method for preparing a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to claim 4, wherein the mass content of each component in the traditional Chinese medicine is as follows: 35-40% of mugwort leaf, 20-25% of pepper stem and leaf, 15-20% of dandelion, 10-12% of honeysuckle, 4-8% of dried orange peel and 4-7% of liquorice.

7. The method for preparing a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to claim 4, wherein the basic nutrition is a composition of soybean meal and peanut shell, and the soybean meal is: the mass ratio of peanut shells is = (9-11) = (5-7).

8. The method for preparing a complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to any one of claims 4 to 7, wherein the preparation method comprises the following steps:

preparing aspergillus oryzae seed liquid: inoculating Aspergillus oryzae into sterile water for culturing to obtain Aspergillus oryzae seed liquid;

solid fermentation primary culture: preparing the basic nutrients and the traditional Chinese medicines into powder, taking the powder as a culture medium, adding the aspergillus oryzae seed liquid, uniformly mixing to obtain a fermentation mixture, and carrying out fermentation culture;

turning and feeding: supplementing a carbon source and a nitrogen source in the fermentation culture process, turning over, and controlling the water content of the fermentation mixture;

and after fermentation culture is completed, the compound enzyme preparation is prepared.

9. The method for preparing the complex enzyme preparation of aspergillus oryzae fermented traditional Chinese medicine according to claim 8, which is characterized in that,

In the preparation of the aspergillus oryzae seed liquid, inoculating aspergillus oryzae into sterile water, and culturing for 4-8 hours in a constant temperature incubator at 30-35 ℃ to obtain the aspergillus oryzae seed liquid;

During the primary culture of the solid fermentation, the basic nutrition and the traditional Chinese medicine are put into a pulverizer for pulverizing and sieving to obtain powder, the powder is taken as a culture medium, the aspergillus oryzae seed liquid is added, sterile water is supplemented, the mixture is stirred uniformly to obtain a fermentation mixture, and then the fermentation mixture is fermented and cultured in an environment of 30-35 ℃; in the fermentation mixture which is preliminarily prepared, the mass of the aspergillus oryzae is 0.3-0.7% of the mass of the culture medium, and the mass ratio of the culture medium to the total amount of sterile water is (4-6): 3;

Among the turning and feeding: the carbon source is glucose, and the nitrogen source is ammonium sulfate; according to the percentages of carbon sources and nitrogen sources relative to the mass of the culture medium, 0.4-0.6% of glucose and 0.4-0.6% of ammonium sulfate are supplemented on days 2-4, 0.4-0.6% of glucose and 0.4-0.6% of ammonium sulfate are supplemented on days 5-7, and 0.1-0.3% of glucose and 0.1-0.3% of ammonium sulfate are supplemented on days 8-10; controlling the water content of the fermentation mixture to be 30% -55%;

after the fermentation mixture is fermented and cultured for 11-13 days, the water content of the fermentation mixture is controlled to be less than or equal to 10%, and the fermentation mixture is crushed to obtain the compound enzyme preparation.

10. Use of a complex enzyme preparation according to any one of claims 1 to 9 as a feed additive.