CN112655839A - Exogenous enzyme composite preparation and preparation method and application thereof - Google Patents
Exogenous enzyme composite preparation and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of feed additives, and discloses an exogenous enzyme composite preparation, a preparation method and an application thereof, wherein the exogenous enzyme composite preparation comprises the following components: enzymes, probiotics, carriers and protectants; the enzymes comprise acidic cellulase, neutral phytase, acidic xylanase, neutral protease, high-temperature resistant alpha-amylase, neutral mannase, glucose oxidase and pectinase; the probiotics comprise bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio; the carrier is selected from at least one of zeolite powder, montmorillonite or diatomite; the protective agent comprises trehalose, yeast extract and mucopolysaccharide. The exogenous enzyme compound preparation can resist the damage of the physiological environment of the alimentary canal of the raw fish, fully exert the efficacy of the exogenous enzyme, effectively improve the immunity of the raw fish, improve the utilization efficiency of feed nutrients and reduce the pollution of residual feed to aquaculture water.
Description
Technical Field
The invention belongs to the technical field of feed additives, and particularly relates to an exogenous enzyme composite preparation as well as a preparation method and application thereof.
Background
The exogenous enzyme mainly refers to an additional enzyme which is not inherent in the product, and the exogenous enzyme preparation is used for aquatic feed to bring the following effects: 1. directly decompose nutrient substrates, improve the metabolic level and reduce the feed coefficient; 2. exogenous digestive enzyme is supplemented, endogenous digestive enzyme secretion is activated, and the feed utilization rate is improved; 3. eliminating anti-nutritional factors in the feed and improving digestive function; 4. improving immunity and promoting health of aquatic animals; 5. promoting the digestion and absorption of phytate phosphorus and reducing the environmental pollution.
However, most of the current application research of the exogenous enzyme preparation in the aquatic animal feed stays in the qualitative aspect, namely whether the added enzyme preparation has the growth promotion effect on the aquatic animals, whether the feed utilization rate can be improved, the feed coefficient can be reduced, and the phosphorus discharge can be reduced, but the quantitative research on the enzyme preparation is less. In practical application, the effect of applying the same content of specific exogenous enzyme to different aquatic animals is different. This is probably due to the fact that the enzymatic reactions are related to the enzyme systems present in the aquatic animal body and the associated reactions are extremely complex. The enzyme is added and the proper addition amount of the enzyme has different requirements according to different aquatic animals, different raw material types and the mixture ratio, the growth can reach the optimal state only within the proper addition range, and the effect is not good when the enzyme is too low or too high.
Meanwhile, exogenous enzymes can only act after entering the digestive tract of animals along with the feed, so that the enzyme preparation is required to have activity under the physiological conditions of the digestive tract (such as pH value, temperature and ion concentration of the gastrointestinal tract); second, because the enzyme is a protein, the exogenous enzyme must be able to resist degradation by endogenous proteases in the animal's digestive tract.
Therefore, the inventor hopes to develop an exogenous enzyme compound preparation which is particularly suitable for raw fish feed and can fully exert the efficacy of exogenous enzyme aiming at raw fish feed which is one of main nutritional products.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides an exogenous enzyme compound preparation, a preparation method and an application thereof, wherein the exogenous enzyme compound preparation can resist the damage of the physiological environment of the digestive tract of the raw fish and fully exert the efficacy of exogenous enzyme; the immunity of the raw fish can be effectively improved, and the occurrence of diseases is reduced; and anti-nutritional factors in the feed are eliminated, the digestive function is improved, the utilization efficiency of feed nutrients is improved, and the pollution of residual feed to the aquaculture water body is reduced.
An exogenous enzyme complex preparation comprises the following components: enzymes, probiotics, carriers and protectants;
the enzymes comprise acidic cellulase, neutral phytase, acidic xylanase, neutral protease, high-temperature resistant alpha-amylase, neutral mannase, glucose oxidase and pectinase;
the probiotics comprise bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio;
the carrier is at least one of zeolite powder, montmorillonite or diatomite;
the protective agent comprises trehalose, yeast extract and mucopolysaccharide.
There are many kinds of exogenous enzymes, each of which can be classified into various specific classes, for example, proteases at least include acid protease and neutral protease. According to the invention, through researching the digestion physiological characteristics of the raw fish and exploring the influence of various enzymes on the raw fish, long-term screening is carried out to finally discover that the acid cellulase, the neutral phytase, the acid xylanase, the neutral protease, the high temperature resistant alpha-amylase, the neutral mannase, the glucose oxidase, the pectinase and other exogenous enzymes have important influence on the growth of the raw fish, and through finding out the specific enzymes most suitable for the raw fish and the optimal dosage thereof, the various enzymes are reasonably compatible and loaded on the carrier, so that the utilization efficiency of the raw fish on nutrient substances and energy is effectively improved, the disease resistance of the raw fish is improved, the nitrogen excretion of the raw fish can be greatly reduced, the eutrophication of the culture water body is reduced, the water body environment is improved, and very obvious economic benefits and social benefits are achieved.
The carrier used in the invention can be used for loading enzymes and probiotics.
By adding some high-temperature resistant probiotics capable of generating exogenous enzymes (protease, lipase, amylase and the like), the high-temperature resistant performance of an exogenous enzyme preparation can be improved, the gastrointestinal microbial balance of raw fish can be adjusted, the beneficial flora in intestinal tracts can be increased, the growth of harmful bacteria can be inhibited, toxins generated by the harmful bacteria can be reduced and eliminated, anti-nutritional factors can be passivated, the intestinal health of fish bodies can be maintained and promoted, and enzymes lost due to the action of digestive tracts can be supplemented. The addition amount of the probiotics is not suitable to be too much, and the addition of too much probiotics can reduce the ingestion rate, the digestibility and the feed utilization rate of the raw fish.
The protective agent can enhance the high temperature resistance of the exogenous enzyme preparation and resist the damage of the digestive tract to the exogenous enzyme.
Through a plurality of feeding tests and laboratory analysis, the complex enzyme preparation is safe and efficient, and the fed raw fish completely meets the food sanitation requirement.
Preferably, the enzyme activity of the enzyme in the exogenous enzyme preparation is: 4000-6000u/g of acidic cellulase, 500-800u/g of neutral phytase, 400-1000u/g of acidic xylanase, 1000-1500u/g of neutral protease, 700-1200u/g of high-temperature resistant alpha-amylase, 200-300u/g of neutral mannanase, 1200-2000u/g of glucose oxidase and 500-800u/g of pectinase. Experiments of the fish bait show that when the enzyme activity of enzyme components is in the range, the growth condition of the raw fish is the most excellent
Preferably, the mass ratio of the probiotics to the carrier to the protective agent in the exogenous enzyme preparation is (0.5-1.2): (6-8): (0.1-0.3).
Preferably, the mass ratio of bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio in the probiotics is (1-2): (0.2-0.6): (0.4-0.8): (1-2).
Preferably, the mass ratio of the trehalose, the yeast extract and the mucopolysaccharide in the protective agent is (2-3): (3-5): (1-2).
Preferably, at least one of magnesium salt or zinc salt is further included in the exogenous enzyme preparation. The addition of inorganic salts such as magnesium salt and zinc salt into the exogenous enzyme can greatly improve the thermal stability of the exogenous enzyme preparation in the digestive tract.
More preferably, the magnesium salt is magnesium chloride and the zinc salt is zinc chloride.
The preparation method of the exogenous enzyme composite preparation comprises the following steps: premixing the exogenous enzyme, the probiotics and the protective agent, adding the carrier, and mixing to obtain the exogenous enzyme composite preparation.
The exogenous enzyme compound preparation is applied to raw fish feed.
Preferably, the application method of the exogenous enzyme compound preparation in the raw fish feed comprises the following steps: and spraying the exogenous enzyme compound preparation on the surface of the raw fish feed.
The raw fish feed comprises the exogenous enzyme compound preparation, and the addition amount of the exogenous enzyme compound preparation in the raw fish feed is 0.5-2 wt%. When the addition amount of the exogenous enzyme compound preparation is 0.5-2% of the total weight of the raw fish feed, the effect generated by the raw fish is optimal; when added in excess, the amount of other nutrients (protein, starch and lipids) added to the raw fish feed is reduced.
Compared with the prior art, the invention has the following beneficial effects:
the exogenous enzyme composite preparation has excellent performance and is particularly suitable for raw fish feed. The exogenous enzyme compound preparation is added into the raw fish feed, so that the feed coefficient can be reduced by 10-30%, the growth speed of the raw fish using the exogenous enzyme compound preparation in the raw fish feed can be improved by 10-25%, the feed cost can be greatly reduced, the morbidity and the survival rate can be reduced, the breeding cost and the pollution of excrement and residual bait to the environment can be reduced, and the feed is convenient to use, green and environment-friendly and free of residues. According to the invention, the carrier and the protective agent are added into the exogenous enzyme compound preparation, so that the loss of exogenous enzyme in the physiological environment of the digestive tract is less, and meanwhile, by adding a certain amount of high-temperature-resistant probiotics, the active enzyme generated by the high-temperature-resistant probiotics can also compensate the enzyme activity lost due to the damage of the digestive tract.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.
The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.
Example 1
The embodiment provides an exogenous enzyme complex preparation, which comprises the following components: enzymes, probiotics, carriers and protectants;
wherein the enzyme activity of the enzymes in the exogenous enzyme preparation is as follows: 4000u/g of acidic cellulase, 500u/g of neutral phytase, 400u/g of acidic xylanase, 1000u/g of neutral protease, 700u/g of high-temperature-resistant alpha-amylase, 200u/g of neutral mannase, 1200u/g of glucose oxidase and 500u/g of pectinase;
wherein the mass ratio of the probiotics to the carrier to the protective agent is 0.5: 6: 0.1;
wherein the mass ratio of bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio in the probiotics is 1: 0.2: 0.4: 1;
wherein the mass ratio of trehalose, yeast extract and mucopolysaccharide in the protective agent is 3: 5: 2;
the carrier is selected from zeolite powder;
the preparation method of the exogenous enzyme composite preparation in the embodiment comprises the following steps: premixing the exogenous enzyme, the probiotics and the protective agent, adding the carrier, and mixing to obtain the exogenous enzyme composite preparation.
Example 2
The embodiment provides an exogenous enzyme complex preparation, which comprises the following components: enzymes, probiotics, carriers and protectants;
wherein the enzyme activity of the enzymes in the exogenous enzyme preparation is as follows: 6000u/g of acidic cellulase, 800u/g of neutral phytase, 1000u/g of acidic xylanase, 1500u/g of neutral protease, 1200u/g of high-temperature resistant alpha-amylase, 300u/g of neutral mannase, 2000u/g of glucose oxidase and 800u/g of pectinase;
wherein the mass ratio of the probiotics to the carrier to the protective agent is 1.2: 8: 0.3;
wherein the mass ratio of bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio in the probiotics is 2: 0.6: 0.8: 2;
wherein the mass ratio of the trehalose to the yeast extract to the mucopolysaccharide in the protective agent is 2: 3: 1;
the carrier is selected from zeolite powder;
the preparation method of the exogenous enzyme composite preparation in the embodiment comprises the following steps: premixing the exogenous enzyme, the probiotics and the protective agent, adding the carrier, and mixing to obtain the exogenous enzyme composite preparation.
Example 3
This example provides a complex formulation of exogenous enzymes, and compared with example 1, example 3 is different in that: magnesium chloride and zinc chloride which account for 1 percent of the total mass of the exogenous enzyme compound preparation are also added (the mass ratio of the magnesium chloride to the zinc chloride is 1: 1);
the preparation method of the exogenous enzyme composite preparation in the embodiment comprises the following steps: premixing the exogenous enzyme, the probiotics, the protective agent, the magnesium chloride and the zinc chloride, adding the carrier, and mixing to obtain the exogenous enzyme composite preparation.
Comparative example 1
The present comparative example provides an exogenous enzyme complex formulation, and compared to example 1, comparative example 1 is distinguished in that: wherein the enzyme activity of the enzymes in the exogenous enzyme preparation is as follows: 4000u/g of neutral cellulase, 800u/g of neutral phytase, 400u/g of acidic xylanase, 1000u/g of acidic protease, 700u/g of high-temperature-resistant alpha-amylase, 200u/g of neutral mannanase and 1200u/g of glucose oxidase;
comparative example 2
This comparative example provides an exogenous enzyme complex formulation, and compared to example 1, comparative example 2 is distinguished in that: wherein the probiotics only comprise bacillus subtilis and bacillus licheniformis, and the mass ratio of the bacillus subtilis to the bacillus licheniformis is 1: 1.
Comparative example 3
The present comparative example provides an exogenous enzyme complex formulation, and compared to example 1, comparative example 3 is distinguished in that: wherein the protectant comprises trehalose.
Comparative example 4
This comparative example provides an exogenous enzyme complex formulation, and compared to example 1, comparative example 4 is distinguished in that: no carrier is contained.
Product effectiveness testing
The exogenous enzyme complex preparations prepared in examples 1-3 and comparative examples 1-4 were respectively sprayed on conventional raw fish feed (25% of bean cake, 10% of chicken meal, 25% of fish meal, 15% of bran, 15% of flour, 6% of fish oil, 2% of vitamin complex, 1% of inorganic salt and 1% of feed adhesive), and the amount of the exogenous enzyme complex preparation added was 1% of the total weight of the raw fish feed, to prepare corresponding raw fish feed products.
The growth and cultivation tests were carried out by feeding raw fish, which were the species of aquatic animals used in the cultivation tests, with the raw fish feeds prepared in examples 1 to 3 and comparative examples 1 to 4, and with a conventional raw fish feed. Adopting a circulating water culture system to carry out a growth culture test, randomly dividing 400 fries of raw fish which are purchased from the market and have basically the same growth condition into 8 groups, and carrying out a growth culture test on 50 fishes in each group under the environmental conditions of the same temperature, pH value and the like for 60 days; the feeding method and the culture condition are both conventional methods and conditions. After the culture test is finished, the weight gain rate, the feed coefficient, the disease incidence rate and the survival rate of the raw fish are measured, and the test results are shown in table 1:
TABLE 1 raw fish culture test results
As shown in Table 1, the performance of the raw fish feed supplemented with the exogenous enzyme complex formulations prepared in examples 1 to 3 and comparative examples 1 to 4 was improved as compared with the conventional raw fish feed. Compared with the exogenous enzyme compound preparation in the comparative examples 1-4, the exogenous enzyme compound preparation in the examples 1-3 is added into the raw fish feed, so that the indexes of the fed raw fish, such as the weight gain rate, the feed coefficient, the disease incidence rate, the survival rate and the like, are obviously improved, and the exogenous enzyme compound preparation product with excellent performance can be prepared only under the selection conditions of proper enzymes, protective agents, probiotics and carriers. It can also be seen from table 1 that, compared with examples 1-2, the addition of magnesium chloride and zinc chloride in example 3 further improves the stability of the complex preparation of exogenous enzyme in the digestive tract of raw fish, thereby ensuring the full exertion of the efficacy of exogenous enzyme.
Claims (10)
1. The exogenous enzyme composite preparation is characterized by comprising the following components: enzymes, probiotics, carriers and protectants;
the enzymes comprise acidic cellulase, neutral phytase, acidic xylanase, neutral protease, high-temperature resistant alpha-amylase, neutral mannase, glucose oxidase and pectinase;
the probiotics comprise bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio;
the carrier is at least one of zeolite powder, montmorillonite or diatomite;
the protective agent comprises trehalose, yeast extract and mucopolysaccharide.
2. The exogenous enzyme complex formulation according to claim 1, wherein the enzyme activity of the enzyme class in the exogenous enzyme formulation is: 4000-6000u/g of acidic cellulase, 500-800u/g of neutral phytase, 400-1000u/g of acidic xylanase, 1000-1500u/g of neutral protease, 700-1200u/g of high-temperature resistant alpha-amylase, 200-300u/g of neutral mannanase, 1200-2000u/g of glucose oxidase and 500-800u/g of pectinase.
3. The exogenous enzyme composite preparation according to claim 1, wherein the mass ratio of the probiotics to the carrier to the protective agent in the exogenous enzyme preparation is (0.5-1.2): (6-8): (0.1-0.3).
4. The exoenzyme complex formulation according to claim 1, wherein the mass ratio of bacillus subtilis, bacillus licheniformis, streptococcus faecalis and bdellovibrio in the probiotic is (1-2): (0.2-0.6): (0.4-0.8): (1-2).
5. The complex formulation of exogenous enzyme according to claim 1, wherein the mass ratio of trehalose, yeast extract and mucopolysaccharide in the protective agent is (2-3): (3-5): (1-2).
6. The complex exogenous enzyme preparation according to claim 1, further comprising at least one of a magnesium salt or a zinc salt.
7. A method for preparing the complex formulation of exogenous enzymes according to any of claims 1 to 5, characterized in that it comprises the following steps: premixing the exogenous enzyme, the probiotics and the protective agent, adding the carrier, and mixing to obtain the exogenous enzyme composite preparation.
8. Use of the complex formulation of exogenous enzymes according to any one of claims 1 to 6 in raw fish feed.
9. The use of claim 8, wherein the exogenous enzyme complex preparation is applied to raw fish feed by the following method: and spraying the exogenous enzyme compound preparation on the surface of the raw fish feed.
10. A raw fish feed comprising the exogenous enzyme complex formulation of any one of claims 1 to 6, wherein the exogenous enzyme complex formulation is added to the raw fish feed in an amount of 0.5 to 2 wt%.
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