CN114831212A - Complex enzyme preparation for enzymolysis of hoof and horn and application thereof - Google Patents
Complex enzyme preparation for enzymolysis of hoof and horn and application thereof Download PDFInfo
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- CN114831212A CN114831212A CN202210404037.5A CN202210404037A CN114831212A CN 114831212 A CN114831212 A CN 114831212A CN 202210404037 A CN202210404037 A CN 202210404037A CN 114831212 A CN114831212 A CN 114831212A
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 113
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 113
- 210000000003 hoof Anatomy 0.000 title claims abstract description 95
- 238000002360 preparation method Methods 0.000 title claims abstract description 64
- 229940088598 enzyme Drugs 0.000 claims abstract description 112
- 239000000843 powder Substances 0.000 claims abstract description 72
- 239000004365 Protease Substances 0.000 claims abstract description 48
- 108091005804 Peptidases Proteins 0.000 claims abstract description 18
- 235000019419 proteases Nutrition 0.000 claims abstract description 18
- 102000004882 Lipase Human genes 0.000 claims abstract description 16
- 108090001060 Lipase Proteins 0.000 claims abstract description 16
- 239000004367 Lipase Substances 0.000 claims abstract description 16
- 235000019421 lipase Nutrition 0.000 claims abstract description 16
- 102000004400 Aminopeptidases Human genes 0.000 claims abstract description 15
- 108090000915 Aminopeptidases Proteins 0.000 claims abstract description 15
- 108010004032 Bromelains Proteins 0.000 claims abstract description 15
- 108090000526 Papain Proteins 0.000 claims abstract description 15
- 239000003674 animal food additive Substances 0.000 claims abstract description 15
- 230000001580 bacterial effect Effects 0.000 claims abstract description 15
- 235000019835 bromelain Nutrition 0.000 claims abstract description 15
- 108010059345 keratinase Proteins 0.000 claims abstract description 15
- 229940055729 papain Drugs 0.000 claims abstract description 15
- 235000019834 papain Nutrition 0.000 claims abstract description 15
- 241001465754 Metazoa Species 0.000 claims abstract description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 3
- 230000000694 effects Effects 0.000 claims description 58
- 230000007071 enzymatic hydrolysis Effects 0.000 claims 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims 1
- 235000019621 digestibility Nutrition 0.000 abstract description 25
- 102000004169 proteins and genes Human genes 0.000 abstract description 23
- 108090000623 proteins and genes Proteins 0.000 abstract description 23
- 150000001875 compounds Chemical class 0.000 abstract description 14
- 238000000338 in vitro Methods 0.000 abstract description 11
- 238000000034 method Methods 0.000 abstract description 8
- 239000002994 raw material Substances 0.000 abstract description 8
- 239000002699 waste material Substances 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 229940079919 digestives enzyme preparation Drugs 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 235000018102 proteins Nutrition 0.000 description 22
- 238000012360 testing method Methods 0.000 description 18
- 102000035195 Peptidases Human genes 0.000 description 15
- 241001494479 Pecora Species 0.000 description 8
- 241000283690 Bos taurus Species 0.000 description 7
- 235000019733 Fish meal Nutrition 0.000 description 6
- 230000001079 digestive effect Effects 0.000 description 6
- 239000004467 fishmeal Substances 0.000 description 6
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 102000057297 Pepsin A Human genes 0.000 description 4
- 108090000284 Pepsin A Proteins 0.000 description 4
- 235000019764 Soybean Meal Nutrition 0.000 description 4
- 102000004142 Trypsin Human genes 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 229940111202 pepsin Drugs 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000004455 soybean meal Substances 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 235000012054 meals Nutrition 0.000 description 3
- 238000007873 sieving Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 108010010803 Gelatin Proteins 0.000 description 2
- 108010076876 Keratins Proteins 0.000 description 2
- 102000011782 Keratins Human genes 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 229920000832 Cutin Polymers 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 235000019735 Meat-and-bone meal Nutrition 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 238000004134 energy conservation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 210000003746 feather Anatomy 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 108010091431 meat tenderizer Proteins 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000007065 protein hydrolysis Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/26—Animal feeding-stuffs from material of animal origin from waste material, e.g. feathers, bones or skin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Biomedical Technology (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Fodder In General (AREA)
Abstract
The invention belongs to the technical field of enzyme preparations, and particularly provides a compound enzyme preparation for hoof and horn enzymolysis and application thereof, wherein the compound enzyme preparation comprises the following components in percentage by mass: 10-15% of bacterial protease, 20-30% of keratinase, 10-15% of arazyme, 5-10% of papain, 5-10% of bromelain, 20-25% of aminopeptidase and 10-15% of lipase. The hoof and horn powder is subjected to enzymolysis by using the compound enzyme preparation, so that on one hand, the waste utilization of hoofs and horns is realized, the feed resource is saved, and the environmental protection pressure is relieved; on the other hand, the method reduces uncertain influence caused by unstable quality, improves the in vitro digestibility of the hoof and horn powder, expands the variety of unconventional feed raw materials and provides scientific basis for substituting protein raw materials. The prepared hoof and horn powder feed additive is applied to animal feed, so that the feed quality can be improved, the feed utilization rate is improved, the feed cost is reduced, and good economic benefit is obtained.
Description
Technical Field
The invention belongs to the technical field of enzyme preparations, and particularly relates to a compound enzyme preparation for hoof and horn enzymolysis and application thereof.
Background
In recent years, the price of the bean pulp rises dramatically all the way, the price fluctuation of the protein raw materials is normalized, and in order to meet the urgent needs of promoting the weight gain of livestock and poultry, producing milk and egg-laying animal feed, the compound feed is produced by purchasing fish meal or meat and bone meal. The price of the waste hoof and horn of the slaughterhouse is far lower than that of the bean pulp and the imported fish meal, and the hoof and horn meal is applied to the feed, so that feed enterprises can be effectively helped to improve the feed quality and reduce the cost, and meanwhile, energy conservation and emission reduction can be realized.
The hoof and horn powder is made up by using hoofs and horns of pig, cow, sheep and horse, and contains rich phosphorus, calcium and several trace elements, and can be used as a high-quality protein feed, and its protein quality is only second to that of fish powder. The quality of protein varies greatly with the raw materials, generally, the meat meal and the hoof and horn meal contain more connective tissue protein and poor amino acid composition, the energy mainly comes from protein and fat (8-18%), and the content and quality of the two components are greatly influenced by the types and processing technology of the raw materials, so that the energy content varies greatly and cannot be fully utilized.
Therefore, the invention provides a compound enzyme preparation, hoof and horn powder is produced by a standard production process, and a feed additive produced by the compound enzyme preparation can be used as an effective substitute resource for soybean meal and fish meal.
Disclosure of Invention
The invention aims to solve the problems of high cost and resource waste in the prior art.
Therefore, the invention provides a compound enzyme preparation for enzymolysis of hoof and horn, which comprises the following components in percentage by mass: 10-15% of bacterial protease, 20-30% of keratinase, 10-15% of arazyme, 5-10% of papain, 5-10% of bromelain, 20-25% of aminopeptidase and 10-15% of lipase.
Specifically, the enzyme activity of the bacterial protease is 300000-500000U/g, the enzyme activity of the keratinase is 100000-300000U/g, the enzyme activity of the arazyme is 100000-300000U/g, the enzyme activity of the papain is 400000-600000U/g, the enzyme activity of the bromelain is 200000-700000U/g, the enzyme activity of the aminopeptidase is 100000-350000U/g, and the enzyme activity of the lipase is 200000-500000U/g.
The invention also provides a feed additive prepared by mixing the complex enzyme preparation and hoof and horn powder, wherein 0.1-0.5 g of complex enzyme preparation is added into every kilogram of hoof and horn powder. Can improve the dry matter digestibility of hoof and horn powder by 0.14-2.77% and the protein digestibility by 1.01-9.53%.
The feed additive can be used as an effective substitute resource of soybean meal and fish meal in feed, and the addition amount is preferably 2-3% of the basic daily ration.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the complex enzyme preparation provided by the invention belongs to a green safe biological additive, accords with the modern food consumption concept, and solves the problems of undefined type and unqualified effect data of the existing feed additive. The hoof and horn powder is subjected to enzymolysis by using the compound enzyme preparation, so that on one hand, the waste utilization of hoofs and horns is realized, the feed resource is saved, and the environmental protection pressure is relieved; on the other hand, the method reduces uncertain influence caused by unstable quality, improves the in vitro digestibility of the hoof and horn powder, expands the variety of unconventional feed raw materials and provides a scientific basis for replacing protein raw materials. The prepared hoof and horn powder feed additive is applied to animal feed, so that the feed quality can be improved, the feed utilization rate is improved, the adverse effect caused by the rising of the prices of raw materials such as soybean meal and fish meal is reduced, the feed cost is reduced, and good economic benefit is obtained.
Detailed Description
The technical solutions in the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Although representative embodiments of the present invention have been described in detail, those skilled in the art to which the present invention pertains will appreciate that various modifications and changes can be made to the present invention without departing from the scope of the invention. Therefore, the scope of the present invention should not be limited to the embodiments, but should be defined by the appended claims and equivalents thereof.
The invention provides a compound enzyme preparation for hoof and horn enzymolysis, which comprises the following components in percentage by mass: 10-15% of bacterial protease, 20-30% of keratinase, 10-15% of arazyme, 5-10% of papain, 5-10% of bromelain, 20-25% of aminopeptidase and 10-15% of lipase. Wherein the enzyme activity of the bacterial protease is 300000-500000U/g, the enzyme activity of the keratinase is 100000-300000U/g, the enzyme activity of the arazyme is 100000-300000U/g, the enzyme activity of the papain is 400000-600000U/g, the enzyme activity of the bromelain is 200000-700000U/g, the enzyme activity of the aminopeptidase is 100000-350000U/g, and the enzyme activity of the lipase is 200000-500000U/g.
The bacterial protease has wide sources, and can improve the utilization rate and the conversion rate of the feed protein. Keratinases specifically degrade keratin, both soluble (e.g., casein, gelatin, etc.) and insoluble proteins (feathers, cutin, nails, etc.). Arazyme can significantly degrade keratin, collagen, elastin, gelatin and the like, preferentially hydrolyze peptide bonds formed by carboxyl ends of arginine, phenylalanine and the like; has strong resistance to pepsin and trypsin. Papain and bromelain are mild protease preparations with high stability, which can hydrolyze macromolecular proteins into small molecular peptides or amino acids, and can be made into meat tenderizer, hydrolyzed sheep placenta extract, etc. The aminopeptidase and the protease are used in a compound way, so that the protein enzymolysis liquid can be further hydrolyzed, the bitter taste of the protein enzymolysis liquid is removed, and the protein hydrolysis degree and the polypeptide content can be greatly improved. The energy of the hoof and horn powder mainly comes from protein and fat, and lipase can act together with protease to catalyze and decompose oil and fat substances, so that the protein digestibility is further improved.
The invention also provides a feed additive prepared by mixing the complex enzyme preparation and hoof and horn powder, wherein 0.1-0.5 g of complex enzyme preparation is added into every kilogram of hoof and horn powder. Can improve the dry matter digestibility of hoof and horn powder by 0.14-2.77% and the protein digestibility by 1.01-9.53%.
The feed additive can be used as an effective substitute resource of soybean meal and fish meal in feed, and the addition amount is preferably 2-3% of the basic daily ration.
The complex enzyme preparation and the effect thereof in the enzymolysis of hoof and horn powder are studied by specific examples.
Example 1:
the embodiment provides a compound enzyme preparation for enzymolysis of hoof and horn, which comprises the following components in percentage by mass: 13% of bacterial protease, 25% of keratinase, 12% of arazyme, 8% of papain, 8% of bromelain, 22% of aminopeptidase and 12% of lipase. Wherein the enzyme activity of the bacterial protease is 300000U/g, the enzyme activity of the keratinase is 200000U/g, the enzyme activity of the arazyme is 200000U/g, the enzyme activity of the papain is 500000U/g, the enzyme activity of the bromelain is 400000U/g, the enzyme activity of the aminopeptidase is 200000U/g, and the enzyme activity of the lipase is 300000U/g.
Example 2:
the embodiment provides a compound enzyme preparation for enzymolysis of hoof and horn, which comprises the following components in percentage by mass: 10% bacterial protease, 20% keratinase, 10% arachnidase, 10% papain, 10% bromelain, 25% aminopeptidase, 15% lipase. Wherein the enzyme activity of the bacterial protease is 300000U/g, the enzyme activity of the keratinase is 200000U/g, the enzyme activity of the arazyme is 200000U/g, the enzyme activity of the papain is 500000U/g, the enzyme activity of the bromelain is 400000U/g, the enzyme activity of the aminopeptidase is 200000U/g, and the enzyme activity of the lipase is 300000U/g.
Example 3:
the embodiment provides a compound enzyme preparation for enzymolysis of hoof and horn, which comprises the following components in percentage by mass: 15% of bacterial protease, 30% of keratinase, 15% of arazyme, 5% of papain, 5% of bromelain, 20% of aminopeptidase and 10% of lipase. Wherein the enzyme activity of the bacterial protease is 300000U/g, the enzyme activity of the keratinase is 200000U/g, the enzyme activity of the arazyme is 200000U/g, the enzyme activity of the papain is 500000U/g, the enzyme activity of the bromelain is 400000U/g, the enzyme activity of the aminopeptidase is 200000U/g, and the enzyme activity of the lipase is 300000U/g.
Comparative example 1:
the comparative example provides a complex enzyme preparation, which is different from the example 1 in that the complex enzyme preparation of the comparative example does not contain lipase, and the mass ratio of the other enzymes is the same as that of the example 1. Wherein the enzyme activity of the bacterial protease is 300000U/g, the enzyme activity of the keratinase is 200000U/g, the enzyme activity of the arazyme is 200000U/g, the enzyme activity of the papain is 500000U/g, the enzyme activity of the bromelain is 400000U/g, and the enzyme activity of the aminopeptidase is 200000U/g.
Example 4:
the method comprises the following steps of crushing cattle and sheep hoof horns, sieving the crushed cattle and sheep hoof horns with a 60-mesh sieve to obtain hoof horn powder, and averagely dividing the hoof horn powder into 5 test groups and 3 control groups, wherein:
control group 1: the hoof and horn powder is not added with any enzyme preparation;
control group 2: adding the complex enzyme preparation provided in comparative example 1 into hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of hoof and horn powder;
control group 3: adding lipase with enzyme activity of 300000U/g into the hoof and horn powder at the addition of 0.5 g/kg;
test group 1: adding the complex enzyme preparation provided in the embodiment 1 into hoof and horn powder according to the proportion that 0.1g of the complex enzyme preparation is added into each kilogram of hoof and horn powder;
test group 2: adding the complex enzyme preparation provided in the embodiment 1 into hoof and horn powder according to the proportion that 0.2g of the complex enzyme preparation is added into each kilogram of hoof and horn powder;
test group 3: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of the hoof and horn powder;
test group 4: adding the complex enzyme preparation provided in the embodiment 2 into the hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of the hoof and horn powder;
test group 5: adding the complex enzyme preparation provided in the embodiment 3 into the hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of the hoof and horn powder;
the in-vitro digestibility of each group of hydrolyzed hoof and horn powder is respectively detected by adopting an in-vitro enzymolysis test, and the method comprises the following specific steps: taking 1g of the hoof and horn powder after enzymolysis at 41 ℃, adding 8mL of buffer solution with the pH value of 2.0 as a substrate, adding 2mL of pepsin (sigma P7012) with the concentration of 2mg/mL, digesting for 45min in a conical flask, transferring the digestive juice into a 8000-plus 14000kD dialysis bag, adjusting the pH value of the digestive juice to 6.5, adding 1.5mL of trypsin (sigma P7545) solution with the concentration of 1.5mg/mL, and digesting for 4h in a constant-temperature water bath shaker at 41 ℃.
The water content and the protein content of each group after digestion were measured, and the results are shown in table 1.
TABLE 1 Effect of Complex enzyme preparation on in vitro digestibility of hoof and horn powders
It can be known from the results of the test groups 1-5 and the control group 1 in table 1 that the hoof and horn powder dry matter digestibility and the protein digestibility can be improved by adding the complex enzyme preparation provided by the invention, and when the adding amount is 0.1g/kg, 0.2g/kg and 0.5g/kg, the dry matter digestibility can be respectively improved by 0.07%, 2.99% and 7.07%, and the protein digestibility can be respectively improved by 2.55%, 5.07% and 16.40%; the complex enzyme preparations provided in examples 2 and 3 with the addition amount of 0.5g/kg can respectively improve the dry matter digestibility by 6.96% and 5.36%, and the protein digestibility by 15.71% and 12.42%. Compared with the control group 1 without the enzyme preparation, the control group 2 and the control group 3 have slightly improved dry matter digestibility and protein digestibility of the hoof powder, but the effect is far lower than that of the complex enzyme preparation provided by the invention.
Example 5:
the application effect of the complex enzyme preparation in the feed is researched.
The method comprises the following steps of crushing cattle and sheep hoof horns, sieving the crushed cattle and sheep hoof horns with a 60-mesh sieve to obtain hoof horn powder, and averagely dividing the hoof horn powder into 3 test groups and 1 control group, wherein:
control group 1: the hoof and horn powder is not added with any enzyme preparation; adding hoof and horn powder into basic ration in an addition amount of 2%;
test group 1: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.1g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into basic daily ration in an addition amount of 2%;
test group 2: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.2g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into basic daily ration by 2 percent of adding amount;
test group 3: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into the basic daily ration by 2 percent of adding amount.
In-vitro enzymolysis tests are adopted to respectively detect the in-vitro digestibility of each group of basic ration added with hoof and horn powder, and the method comprises the following specific steps: taking 1g of basal ration added with hoof and horn powder into a conical flask at 41 ℃, adding 8mL of buffer solution with the pH value of 2.0 as a substrate, adding 2mL of pepsin (sigma P7012) with the concentration of 2mg/mL, digesting for 45min in the conical flask, transferring the digestive juice into a 8000-14000kD dialysis bag, adjusting the pH value of the digestive juice to 6.5, adding 1.5mL of trypsin (sigma P7545) solution with the concentration of 1.5mg/mL, and digesting for 4h in a constant-temperature water bath shaker at 41 ℃.
The water content and the protein content of each group after digestion were measured, and the results are shown in table 2.
TABLE 2 Effect of Complex enzyme preparation on feed digestibility in vitro
The results in table 2 show that the feed additive prepared by adding the complex enzyme preparation provided by the invention into the hoof and horn powder in the addition amount of 0.1g/kg, 0.2g/kg and 0.5g/kg can respectively improve the dry matter digestibility by 0.14%, 1.24% and 2.77% and the protein digestibility by 1.01%, 0.69% and 9.53% when the feed additive is used in feed.
Example 6:
the application effect of the complex enzyme preparation in the feed is researched.
The method comprises the following steps of crushing cattle and sheep hoof horns, sieving the crushed cattle and sheep hoof horns with a 60-mesh sieve to obtain hoof horn powder, and averagely dividing the hoof horn powder into 3 test groups and 1 control group, wherein:
control group 1: the hoof and horn powder is not added with any enzyme preparation; adding hoof and horn powder into basic ration by 3%;
test group 1: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.1g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into basic daily ration by 3 percent of adding amount;
test group 2: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.2g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into basic daily ration by 3 percent of adding amount;
test group 3: adding the complex enzyme preparation provided in the embodiment 1 into the hoof and horn powder according to the proportion of adding 0.5g of the complex enzyme preparation into each kilogram of the hoof and horn powder; adding the hoof and horn powder after enzymolysis into the basic daily ration by 3 percent of adding amount.
In-vitro enzymolysis tests are adopted to respectively detect the in-vitro digestibility of each group of basic ration added with the hoof and horn powder, and the method comprises the following specific steps: taking 1g of basal ration added with hoof and horn powder into a conical flask at 41 ℃, adding 8mL of buffer solution with the pH value of 2.0 as a substrate, adding 2mL of pepsin (sigma P7012) with the concentration of 2mg/mL, digesting for 45min in the conical flask, transferring the digestive juice into a 8000-14000kD dialysis bag, adjusting the pH value of the digestive juice to 6.5, adding 1.5mL of trypsin (sigma P7545) solution with the concentration of 1.5mg/mL, and digesting for 4h in a constant-temperature water bath shaker at 41 ℃.
The water content and the protein content of each group after digestion were measured, and the results are shown in table 2.
TABLE 3 Effect of Complex enzyme preparation on feed digestibility in vitro
The results in table 3 show that the feed additive prepared by adding the complex enzyme preparation provided by the invention into the hoof and horn powder in the addition amount of 0.1g/kg, 0.2g/kg and 0.5g/kg can respectively improve the dry matter digestibility by 0.08%, 0.72% and 1.03% and the protein digestibility by 0.53%, 0.85% and 1.18% when the feed additive is used in feed.
The above examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention, which is intended to be covered by the claims and any design similar or equivalent to the scope of the invention.
Claims (4)
1. A complex enzyme preparation for enzymolysis of hoofs and horns is characterized by comprising the following components in percentage by mass: 10-15% of bacterial protease, 20-30% of keratinase, 10-15% of arazyme, 5-10% of papain, 5-10% of bromelain, 20-25% of aminopeptidase and 10-15% of lipase.
2. A complex enzyme preparation for enzymatic hydrolysis of hoof and horn according to claim 1, wherein: the enzyme activity of the bacterial protease is 300000-500000U/g, the enzyme activity of the keratinase is 100000-300000U/g, the enzyme activity of the arazyme is 100000-300000U/g, the enzyme activity of the papain is 400000-600000U/g, the enzyme activity of the bromelain is 200000-700000U/g, the enzyme activity of the aminopeptidase is 100000-350000U/g, and the enzyme activity of the lipase is 200000-500000U/g.
3. A feed additive comprising the complex enzyme preparation for enzymolysis of hoof and hoof powder according to any one of claims 1-2, wherein: 0.1-0.5 g of the complex enzyme preparation is added into every kilogram of hoof and horn powder.
4. Use of a feed additive according to claim 3 in animal feed, wherein: the addition amount of the feed additive is 2-3% of that of basic daily ration.
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