Complex enzyme preparation and preparation method thereof

Abstract

The invention discloses a compound enzyme preparation and a preparation method thereof, and relates to the technical field of enzyme preparations. The compound enzyme preparation is at least prepared from the following components in parts by weight: 10 to 30 parts of xylanase, 15 to 35 parts of modified cellulase, 8 to 20 parts of glucanase, 5 to 15 parts of protease, 5 to 18 parts of amylase, 3 to 12 parts of pectinase, 5 to 25 parts of phytase and 15 to 40 parts of modified superoxide dismutase compound microspheres. The compound enzyme preparation provided by the invention is added into daily ration of ruminants, so that the apparent digestibility and growth performance of the ruminants on nutrients can be improved, meanwhile, safer and better meat quality can be obtained, the meat taste is improved, and higher economic value is obtained.

Description

Technical Field

[0001] The present invention relates to the technical field of enzyme preparations, and particularly relates to a compound enzyme preparation and a preparation method thereof. Background Art

[0002] With the increase in the global population, meeting the demand for animal protein products has become a major issue in global food security. Nowadays, the reduction in the area of agricultural land has led to a shortage of feed resources, thus affecting the development of livestock production. How to improve the feed utilization rate of livestock and poultry has become a major problem in current livestock and poultry farming. Since the ban on antibiotics, the addition of compound enzymes to livestock and poultry diets to improve production performance has been widely used in production. As a green and efficient new type of feed additive, enzyme preparations can improve the production performance of livestock and poultry, increase the utilization efficiency of livestock and poultry for feed, reduce pollution, and play an important role in fundamentally improving the production efficiency of beef cattle. They also provide a broader space for the selection of feed raw materials. Enzyme preparations, as feed additives, have now been widely used in poultry farming. Different types of enzyme preparations play their unique roles in feed, which can improve the nutrient utilization rate of feed and the growth performance of poultry: Cellulase and pectinase can effectively break down the plant cell wall and release the nutrients inside the cells, thereby increasing the nutrient utilization rate of feed; β-glucanase and xylanase focus on decomposing the anti-nutritional factors in feed, reducing the anti-nutritional effect of non-starch polysaccharides on poultry, and improving the digestion and absorption efficiency of feed; Protease and amylase can make up for the deficiency of endogenous enzymes in poultry, help poultry digest and utilize protein and starch substances more effectively, and promote growth and development.

[0003] Different enzyme preparations have a synergistic effect. A compound enzyme preparation is a mixture of different types of enzymes in an appropriate ratio, enabling each component to play a better role. Currently, in modern poultry feeding, adding compound enzyme preparations with different ratios has become a common practice. The structure of the cell wall of roughage is complex, and the cellulose and lignin in it produce a "cage" effect, which is not conducive to the digestion and absorption of feed by ruminants. Therefore, in production, it is crucial to break the "cage" effect of cellulose and lignin and improve the utilization rate of roughage for ruminants. The application research of fiber compound enzymes mostly focuses on monogastric animals such as pigs and chickens, and many research results have confirmed that fiber compound enzymes have significant good effects in improving the feed rate and production performance of pigs and chickens, enhancing the digestion and absorption function, improving the nutritional value of feed, regulating the body metabolism, and improving disease resistance. However, the application research of fiber compound enzymes in ruminant feeding started relatively late. Because the fiber decomposition activity in the rumen is very active, simply adding exogenous enzyme preparations will not increase the fiber decomposition activity. Therefore, existing compound enzyme preparations have defects such as being easily decomposed and inactivated and having poor digestion-promoting effects, which severely limit the use of this technology. Summary of the Invention

[0004] The object of the present invention is to provide a compound enzyme preparation and a preparation method thereof, and solve the following technical problems: When the existing compound enzyme preparation is used in ruminant feed, there are problems of easy decomposition and failure and poor digestion-promoting effect.

[0005] The object of the present invention can be achieved by the following technical solutions: A compound enzyme preparation, at least comprising the following components in parts by weight: 10 - 30 parts of xylanase, 15 - 35 parts of modified cellulase, 8 - 20 parts of glucanase, 5 - 15 parts of protease, 5 - 18 parts of amylase, 3 - 12 parts of pectinase, 5 - 25 parts of phytase, 15 - 40 parts of modified superoxide dismutase composite microspheres.

[0006] As a further scheme of the present invention: the modified cellulase is polyethylene glycol-modified cellulase, and the superoxide dismutase is polydopamine-chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase composite microspheres.

[0007] As a further scheme of the present invention: the preparation method of the modified superoxide dismutase composite microspheres comprises the following steps: Adding the sodium alginate solution of superoxide dismutase into isooctane, then adding an emulsifier, and after emulsifying evenly, adding a calcium chloride solution and isopropanone to obtain a superoxide dismutase core layer; Adding the superoxide dismutase core layer into the acetonitrile solution of poly(lactic-co-glycolic acid), and then adding it into peanut oil to obtain poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules; Adding the poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules into a chitosan solution for incubation to obtain chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules; Adding the chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules into a tris(hydroxymethyl)aminomethane hydrochloride buffer solution, and then adding dopamine hydrochloride to obtain modified superoxide dismutase composite microspheres.

[0008] As a further scheme of the present invention: the mass ratio of the superoxide dismutase core layer to the poly(lactic-co-glycolic acid) is 1:5 - 7.

[0009] As a further scheme of the present invention: the mass ratio of the poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules to the chitosan is 1:0.3 - 1.2.

[0010] As a further scheme of the present invention: the mass ratio of the chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules to the dopamine hydrochloride is 1:0.01 - 0.05.

[0011] As a further solution of the present invention: The preparation method of the modified cellulase at least includes the following steps: Add the polyethylene glycol solution to the cellulase solution, and dry to obtain the modified cellulase.

[0012] As a further solution of the present invention: The degree of polymerization of the polyethylene glycol is 2000-8000, and the mass ratio of the polyethylene glycol to the cellulase is 1-20:100.

[0013] A preparation method of a compound enzyme preparation at least includes the following preparation steps: Add xylanase, modified cellulase, glucanase, protease, amylase, pectinase, phytase and modified superoxide dismutase composite microspheres to a mixer, mix evenly and then press into shape to obtain the compound enzyme preparation.

[0014] The beneficial effects of the present invention: The compound enzyme preparation of the present invention at least includes xylanase, modified cellulase, glucanase, protease, amylase, pectinase, phytase and modified superoxide dismutase composite microspheres. Through the synergistic action of multiple enzymes, adding cellulase, pectinase, xylanase and β-glucanase to the diet can destroy the cell wall structure of plant raw materials, increase the contact area between rumen microorganisms and feed, enhance the rumen fiber digestion ability, improve the digestibility of various substances in the diet, and then improve the feed conversion rate, promote the degradation and absorption of non-starch polysaccharide substances such as cellulose, xylan and pectin, and improve the nutritional value of feed fermentation in the rumen, thereby improving the overall digestion and absorption of animals to feed. At the same time, the exogenous enzyme preparation can also cooperate with the endogenous enzyme to improve the attachment of microorganisms to feed particles and play a role. Adding the prepared compound enzyme preparation to the diet of ruminants in the present invention can effectively improve the activity of endogenous enzymes in the rumen, thereby improving the apparent digestibility of nutrients and growth performance of ruminants.

[0015] In the present invention, the cellulase is physically coated to obtain the modified cellulase. The cellulase coated with polyethylene glycol has enhanced enzyme activity and stability. Through polyethylene glycol coating, the action time of the enzyme can be extended. Polyethylene glycol remains stable in the acidic environment of the rumen and gradually dissolves after entering the intestine, increasing the release rate in the intestine, thereby enhancing the decomposition of cellulose and improving the feed digestibility. Polyethylene glycol forms hydrogen bonds and hydrophobic interactions with the cellulase, which can buffer thermal shock, protect the enzyme spatial structure, and reduce the thermal inactivation of the enzyme during feed processing. At the same time, using polyethylene glycol to physically coat the cellulase has no risk of toxic residue, does not affect the rumen microbial community balance, and the preparation method is simple and the cost is low, which is suitable for large-scale production.

[0016] In the composite enzyme preparation of the present invention, superoxide dismutase is also added, and the superoxide dismutase is prepared into modified superoxide dismutase composite microspheres, forming a four-layer wrapping structure of polydopamine-chitosan-polylactic acid glycolic acid-calcium alginate-superoxide dismutase, which not only protects the activity of superoxide dismutase, but also enables the superoxide dismutase to be slowly released with the degradation of the polymer material, having high stability and high heat resistance. In the rumen stage, the calcium alginate layer and the polylactic acid glycolic acid layer provide the ability for superoxide dismutase to resist the acidic environment and slow release, avoiding the rapid inactivation of superoxide dismutase directly exposed in the rumen and losing its antioxidant function; in the intestinal stage, the calcium alginate layer and the polylactic acid glycolic acid layer degrade, and the chitosan layer and the polydopamine layer provide adhesiveness, improving the absorption rate of superoxide dismutase, thereby improving the feed conversion rate, enhancing the immunity, scavenging harmful free radicals in the body to prevent the occurrence of diseases, and being safe without side effects. In the present invention, the modified superoxide dismutase composite microspheres are prepared into a composite enzyme preparation and then added to the ruminant diet, so as to increase the content of superoxide dismutase in poultry from two aspects of exogenous supplementation and endogenous stimulation in the body, reduce the use of antibiotics, enhance the anti-stress ability, improve nutrient absorption, improve the disease resistance and survival rate of ruminants, and reduce the probability of getting sick. At the same time, safer and better-quality meat poultry can be obtained, the meat texture and taste can be improved, and higher economic value can be obtained. Detailed implementation mode

[0017] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0018] Example 1 The preparation method of the modified superoxide dismutase composite microspheres includes the following steps: 80 mg of superoxide dismutase with an enzyme activity of 30000 U / g is dissolved in a phosphate buffer solution containing 1 wt% sodium alginate, added to isooctane containing 6 wt% span 80 according to a volume ratio of 1:2, homogenized at high speed for 3 min, then tween 80 is added dropwise, homogenized for 3 min, 8 wt% calcium chloride solution is added dropwise and then homogenized for 3 min, and finally, isopropanol is added and homogenized for 3 min at the same rotation speed, centrifuged, washed, and freeze-dried to obtain the superoxide dismutase core layer; Dissolve 400 mg of poly (lactic-co-glycolic acid) in 10 mL of acetonitrile, add the superoxide dismutase core layer prepared above, after ultrasonic treatment with a 100 w ultrasonic probe for 20 s, dropwise add it to peanut oil containing 6 wt% Span 80 under stirring at 600 r / min, then increase the rotation speed to 1000 r / min and continue stirring for 2 min, rotary evaporate under reduced pressure to remove acetonitrile, wash with petroleum ether, and dry at 37 °C to obtain poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules; Add the above poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules to a 0.5 wt% chitosan solution and incubate for 30 min, then centrifuge to collect the microcapsules, wash once with the above chitosan solution and then wash twice with water, and freeze-dry. The mass ratio of poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules to chitosan is 1:0.8 to obtain chitosan-poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules; Add the above chitosan-poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules to a tris (hydroxymethyl) aminomethane hydrochloride buffer solution with a pH of 8.5, then add a dopamine hydrochloride solution with a concentration of 2 mg / mL, magnetically stir at a rotation speed of 300 r / min for 24 h, then add 0.1 mol / L hydrochloric acid solution to adjust the pH to 6, wash and freeze-dry. The mass ratio of chitosan-poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules to dopamine hydrochloride is 1:0.03 to obtain modified superoxide dismutase composite microspheres.

[0019] Example 2 The preparation method of the modified superoxide dismutase composite microspheres includes the following steps: Dissolve 80 mg of superoxide dismutase with an enzyme activity of 30000 U / g in a phosphate buffer solution containing 1 wt% sodium alginate, add it to isooctane containing 6 wt% Span 80 according to a volume ratio of 1:2, homogenize at high speed for 3 min, then dropwise add Tween 80 and homogenize for 3 min, gradually add 8 wt% calcium chloride solution and continue to homogenize for 3 min. Finally, add isopropanol and homogenize at the same rotation speed for 3 min, centrifuge, wash, and freeze-dry to obtain the superoxide dismutase core layer; Dissolve 500 mg of poly (lactic-co-glycolic acid) in 12 mL of acetonitrile, add the superoxide dismutase core layer prepared above, after ultrasonic treatment with a 100 w ultrasonic probe for 20 s, dropwise add it to peanut oil containing 6 wt% Span 80 under stirring at 600 r / min, then increase the rotation speed to 1000 r / min and continue stirring for 2 min, rotary evaporate under reduced pressure to remove acetonitrile, wash with petroleum ether, and dry at 37 °C to obtain poly (lactic-co-glycolic acid)-superoxide dismutase microcapsules; After incubating the above poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules in a 0.8 wt% chitosan solution for 30 min, centrifuging, collecting the microcapsules, washing them once with the above chitosan solution and then twice with water, and freeze-drying them, where the mass ratio of poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules to chitosan is 1:1, chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules are obtained; Add the above chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules to a tris(hydroxymethyl)aminomethane hydrochloride buffer solution with a pH of 8.5, then add a dopamine hydrochloride solution with a concentration of 2 mg / mL, magnetically stir at a speed of 300 r / min for 24 h, then add 0.1 mol / L hydrochloric acid solution to adjust the pH to 6, wash and freeze-dry, where the mass ratio of chitosan-poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules to dopamine hydrochloride is 1:0.03, and modified superoxide dismutase composite microspheres are obtained.

[0020] Example 3 The preparation method of modified cellulase includes the following steps: Add polyethylene glycol with a molecular weight of 6000 to a citric acid-sodium citrate buffer solution with a pH of 4.8 and dissolve it fully to prepare a polyethylene glycol solution with a concentration of 10 g / L, and add the above polyethylene glycol solution to the cellulase solution so that the mass ratio of polyethylene glycol to cellulase is 1:10. The enzyme activity of the cellulase solution is 10 U / mL (specific enzyme activity is 2000 U / g), stir for 0.5 h, and freeze-dry to obtain modified cellulase.

[0021] Example 4 A composite enzyme preparation is made by the following method: Add 20 parts by mass of xylanase with an enzyme activity of 25000 U / g, 25 parts by mass of the modified cellulase prepared in Example 3, 15 parts by mass of glucanase with an enzyme activity of 4500 U / g, 10 parts by mass of protease with an enzyme activity of 5000 U / g, 12 parts by mass of amylase with an enzyme activity of 35000 U / g, 8 parts by mass of pectinase with an enzyme activity of 4200 U / g, 15 parts by mass of phytase with an enzyme activity of 20000 U / g, and 25 parts by mass of the modified superoxide dismutase composite microspheres prepared in Example 1 into a mixer, mix evenly and then press into shape to obtain the composite enzyme preparation.

[0022] Example 5 A composite enzyme preparation is made by the following method: Add 20 parts by mass of xylanase with an enzyme activity of 25,000 U / g, 25 parts by mass of the modified cellulase prepared in Example 3, 15 parts by mass of dextranase with an enzyme activity of 4,500 U / g, 10 parts by mass of protease with an enzyme activity of 5,000 U / g, 12 parts by mass of amylase with an enzyme activity of 35,000 U / g, 8 parts by mass of pectinase with an enzyme activity of 4,200 U / g, 15 parts by mass of phytase with an enzyme activity of 20,000 U / g, and 25 parts by mass of the modified superoxide dismutase composite microspheres prepared in Example 2 into a mixer. After mixing evenly, press and mold to obtain a composite enzyme preparation.

[0023] Example 6 A composite enzyme preparation is made by the following method: Add 25 parts by mass of xylanase with an enzyme activity of 25,000 U / g, 20 parts by mass of the modified cellulase prepared in Example 3, 15 parts by mass of dextranase with an enzyme activity of 4,500 U / g, 10 parts by mass of protease with an enzyme activity of 5,000 U / g, 12 parts by mass of amylase with an enzyme activity of 35,000 U / g, 8 parts by mass of pectinase with an enzyme activity of 4,200 U / g, 15 parts by mass of phytase with an enzyme activity of 20,000 U / g, and 30 parts by mass of the modified superoxide dismutase composite microspheres prepared in Example 1 into a mixer. After mixing evenly, press and mold to obtain a composite enzyme preparation.

[0024] Example 7 A composite enzyme preparation is made by the following method: Add 25 parts by mass of xylanase with an enzyme activity of 25,000 U / g, 20 parts by mass of the modified cellulase prepared in Example 3, 15 parts by mass of dextranase with an enzyme activity of 4,500 U / g, 10 parts by mass of protease with an enzyme activity of 5,000 U / g, 12 parts by mass of amylase with an enzyme activity of 35,000 U / g, 8 parts by mass of pectinase with an enzyme activity of 4,200 U / g, 15 parts by mass of phytase with an enzyme activity of 20,000 U / g, and 30 parts by mass of the modified superoxide dismutase composite microspheres prepared in Example 2 into a mixer. After mixing evenly, press and mold to obtain a composite enzyme preparation.

[0025] Comparative Example 1 The preparation method of the modified superoxide dismutase composite microspheres includes the following steps: Dissolve 80 mg of superoxide dismutase with an enzyme activity of 30,000 U / g in a phosphate buffer solution containing 1 wt% sodium alginate, add it to isooctane containing 6 wt% span 80 according to a volume ratio of 1:2, homogenize at high speed for 3 min, then add tween 80 dropwise and homogenize for 3 min. Gradually add 8 wt% calcium chloride solution and continue to homogenize for 3 min. Finally, add isopropanol and homogenize for 3 min at the same rotation speed, then centrifuge, wash, and freeze-dry to obtain the superoxide dismutase core layer; After incubating the above-mentioned superoxide dismutase nuclear layer in a 0.5 wt% chitosan solution for 30 min, centrifuge to collect the microcapsules. Wash them once with the above chitosan solution and then twice with water, followed by freeze-drying to obtain chitosan-superoxide dismutase microcapsules; Add the above chitosan-superoxide dismutase microcapsules to a tris(hydroxymethyl)aminomethane hydrochloride buffer solution with a pH of 8.5, and then add a dopamine hydrochloride solution with a concentration of 2 mg / mL. Magnetically stir at a speed of 300 r / min for 24 h, and then add 0.1 mol / L hydrochloric acid solution to adjust the pH to 6. Wash and freeze-dry to obtain modified superoxide dismutase composite microspheres.

[0026] Comparative Example 2 The preparation method of the modified superoxide dismutase composite microspheres includes the following steps: Dissolve 80 mg of superoxide dismutase with an enzyme activity of 30000 U / g in a phosphate buffer solution containing 1 wt% sodium alginate, add it to isooctane containing 6 wt% Span 80 according to a volume ratio of 1:2, homogenize at high speed for 3 min, then add Tween 80 and homogenize for 3 min. Dropwise add 8 wt% calcium chloride solution and continue to homogenize for 3 min. Finally, add isopropanol and homogenize at the same speed for 3 min, followed by centrifugation, washing, and freeze-drying to obtain the superoxide dismutase nuclear layer; Dissolve 400 mg of poly(lactic-co-glycolic acid) in 10 mL of acetonitrile, add the above-prepared superoxide dismutase nuclear layer, sonicate with a 100 w ultrasonic probe for 20 s, and then dropwise add it to peanut oil containing 6 wt% Span 80 under stirring at 600 r / min. Subsequently, increase the speed to 1000 r / min and continue to stir for 2 min. Rotate evaporate under reduced pressure to remove acetonitrile, wash with petroleum ether, and dry at 37°C to obtain poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules; After incubating the above poly(lactic-co-glycolic acid)-superoxide dismutase microcapsules in a 0.5 wt% chitosan solution for 30 min, centrifuge to collect the microcapsules. Wash them once with the above chitosan solution and then twice with water, followed by freeze-drying to obtain modified superoxide dismutase composite microspheres.

[0027] Comparative Example 3 Compared with Example 4, in Comparative Example 3, the modified superoxide dismutase composite microspheres prepared in Example 1 added in Example 4 were simply replaced with the modified superoxide dismutase composite microspheres prepared in Comparative Example 1 in equal mass, and the remaining components and preparation methods were exactly the same as those in Example 4.

[0028] Comparative Example 4 Compared with Example 4, in Comparative Example 4, the modified superoxide dismutase composite microspheres prepared in Example 3 added in Example 4 were simply replaced with the modified superoxide dismutase composite microspheres prepared in Comparative Example 2 in equal mass, and the remaining components and preparation methods were exactly the same as those in Example 4.

[0029] Comparative Example 5 Compared with Example 4, in Comparative Example 5, the quality of the modified cellulase prepared in Example 3 was replaced with unmodified cellulase, and the remaining components and preparation method were exactly the same as those in Example 4.

[0030] Comparative Example 6 Compared with Example 4, in Comparative Example 6, the modified superoxide dismutase composite microspheres prepared in Example 1 were not added, and the remaining components and preparation method were exactly the same as those in Example 4.

[0031] Performance detection The Qing goat was provided by Henan Yuanmu Co., Ltd. A total of 200 healthy Qing goats were selected for the experiment, and the average weight was (16.18 ± 0.13) kg. The feed for raising Qing goats met the standard NY / T 816-2004, and the feed composition was 42% corn, 15% wheat bran, 12% soybean meal, 15% peanut cake, 13% alfalfa, 2% salt, and 1% of the composite enzyme preparation prepared in Examples 4-7 and Comparative Examples 3-6. A control group without adding the composite enzyme preparation was set. The 200 Qing goats were randomly divided into 4 groups, with 5 replicates in each group and 10 goats in each replicate. The experiment lasted for 40 days. During the experiment, the Qing goats had free access to water and food, and were immunized according to the conventional immunization program.

[0032] Growth performance test: On the day when the experiment started and ended, the experimental meat goats were fasted for 12 h, and the initial weight and final weight of the experimental meat goats were measured on an empty stomach. The feed intake of each group of meat goats during the experimental period was recorded, and the average daily feed intake, average daily weight gain, and feed-to-weight ratio of each group of meat goats were calculated; Average daily weight gain = (final weight of meat goats in the experiment - initial weight of meat goats in the experiment) / number of experimental days Average daily feed intake = total feed intake / number of experimental days Feed-to-weight ratio = average daily feed intake / average daily weight gain; The test results are shown in Table 1; Apparent digestibility test of nutrients: Three days before the end of the experiment, fresh feces of meat goats were collected continuously every day, 100 g each time, and 10 mL of 10% sulfuric acid was added for nitrogen fixation. The fecal samples of 3 days were mixed correspondingly, dried at 65 °C, pulverized, and passed through a 40-mesh sieve for standby. 200 g of the experimental diets of each group were pulverized and passed through a 40-mesh sieve for standby. The apparent digestibility of crude protein, crude fat, dry matter, acid detergent fiber, and neutral detergent fiber in the diets and feces was measured and calculated by the method of acid-insoluble ash content; Apparent digestibility of a certain nutrient = 100% - (content of this nutrient in feces × acid-insoluble ash content in diet) / (content of this nutrient in diet × acid-insoluble ash content in feces) × 100%; The test results are shown in Table 1; Table 1: Growth and digestion performance test data in Examples 4-7, Comparative Examples 3-6, and the control group

[0033] Immune index test: After the experiment, 2 meat sheep were randomly selected from each replicate, and the jugular vein blood was aseptically collected and placed in a clean centrifuge tube. It was allowed to stand for 15 min and then centrifuged at 2000 r / min for 20 min. The upper serum was aspirated and stored in a -20°C refrigerator for later use. The contents of immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM) were measured using an immunoglobulin ELISA detection kit (Shanghai Enzyme-linked Biotechnology Co., Ltd.). After the meat sheep were slaughtered, their spleens were removed and weighed for calculating the spleen index; Spleen index = fresh spleen weight (g) / meat sheep body weight (kg); The test results are shown in Table 2; Meat quality test: After the experiment, 2 meat sheep were randomly selected from each replicate for slaughter. The right longissimus dorsi muscle was taken, and the meat quality indexes were measured according to the test method in the reference literature. The main indexes measured were the carcass weight, eye muscle area, shear force, cooking loss rate, drip loss rate, and meat color of the meat sheep; The test results are shown in Table 2; Table 2: Detection data of immune performance and meat quality in Examples 4 - 7, Comparative Examples 3 - 6, and the control group

[0034] As can be seen from Table 1 and Table 2, when the compound enzyme preparation prepared by the present invention is added to the diet of ruminants such as sheep, the apparent digestibility of various nutrients is improved, and thus the growth performance is significantly enhanced. Moreover, the addition of the modified superoxide dismutase composite microspheres can improve the immunity and meat quality of Qingshan goats to a certain extent. In Comparative Example 3, the modified superoxide dismutase composite microspheres were added without forming a poly (lactic - glycolic acid) layer, and in Comparative Example 4, the added modified superoxide dismutase composite microspheres did not form a polydopamine layer. When the obtained compound enzyme preparation was used in the diet, the digestion ability of experimental sheep for nutrients, growth performance, immunity indexes, and mutton quality decreased. In Comparative Example 5, the added cellulase was not coated with polyvinyl alcohol, and the digestion ability of experimental sheep for nutrients and growth performance decreased significantly. In Comparative Example 6, the modified superoxide dismutase composite microspheres were not added, and when the obtained compound enzyme preparation was used in the diet, the immunity indexes and mutton quality of mutton decreased significantly.

[0035] The above has described a specific embodiment of the present invention in detail, but the content described is only the preferred embodiment of the present invention and cannot be considered as limiting the scope of implementation of the present invention. All equivalent changes and improvements made according to the scope of the present invention application should still fall within the scope covered by the patent of the present invention.

Claims

1. A composite enzyme preparation, characterized in that, Comprising at least the following components by weight: Xylanase 10 - 30 parts, modified cellulase 15 - 35 parts, glucanase 8 - 20 parts, protease 5 - 15 parts, amylase 5 - 18 parts, pectinase 3 - 12 parts, phytase 5 - 25 parts, modified superoxide dismutase composite microspheres 15 - 40 parts.

2. The composite enzyme preparation according to claim 1, wherein The modified cellulase is polyethylene glycol - modified cellulase, and the superoxide dismutase is polydopamine - chitosan - poly (lactic - co - glycolic acid) - superoxide dismutase composite microspheres.

3. The composite enzyme preparation according to claim 2, characterized in that, The preparation method of the modified superoxide dismutase composite microspheres comprises the following steps: Adding the sodium alginate solution of superoxide dismutase into isooctane, then adding an emulsifier, and after emulsifying evenly, adding a calcium chloride solution and isopropanone to obtain the superoxide dismutase core layer; Adding the superoxide dismutase core layer into the acetonitrile solution of poly (lactic - co - glycolic acid), and then adding it into peanut oil to obtain poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules; Adding the poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules into a chitosan solution for incubation to obtain chitosan - poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules; Adding the chitosan - poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules into a tris - hydrochloride buffer solution, and then adding dopamine hydrochloride to obtain the modified superoxide dismutase composite microspheres.

4. The composite enzyme preparation according to claim 3, wherein The mass ratio of the superoxide dismutase core layer to the poly (lactic - co - glycolic acid) is 1:5 - 7.

5. The composite enzyme preparation according to claim 3, characterized in that, The mass ratio of the poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules to the chitosan is 1:0.3 - 1.

2.

6. The composite enzyme preparation according to claim 3, characterized in that, The mass ratio of the chitosan - poly (lactic - co - glycolic acid) - superoxide dismutase microcapsules to the dopamine hydrochloride is 1:0.01 - 0.

05.

7. A composite enzyme preparation according to claim 1, characterized in that, The preparation method of the modified cellulase comprises at least the following steps: Adding a polyethylene glycol solution into a cellulase solution and drying to obtain the modified cellulase.

8. A composite enzyme preparation according to claim 7, characterized in that, The degree of polymerization of the polyethylene glycol is 2000 - 8000, and the mass ratio of the polyethylene glycol to the cellulase is 1 - 20:

100.

9. A preparation method of a composite enzyme preparation, characterized in that At least comprising the following preparation steps: Adding xylanase, modified cellulase, glucanase, protease, amylase, pectinase, phytase and modified superoxide dismutase composite microspheres into a mixer, mixing evenly and then pressing into shape to obtain the composite enzyme preparation.

Images