CN108486087A - It is a kind of enzymolysis chicken gizzard complex enzyme formulation and its application - Google Patents
It is a kind of enzymolysis chicken gizzard complex enzyme formulation and its application Download PDFInfo
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Abstract
The present invention disclose it is a kind of enzymolysis chicken gizzard complex enzyme formulation and its application, belong to complex enzyme formulation field.The complex enzyme formulation includes the component of following mass parts meter：1~3 part of neutral proteinase, 1~6 part of alkali protease, 1~3 part of lipase, 0.1~0.3 part of visceral enzym, 0.1~0.4 part of food flavor enzyme, 3~5 parts of carrier.The present invention is complex enzyme formulation by different enzyme activity, proportioning, dosage compounding by neutral proteinase, alkali protease, lipase, visceral enzym, food flavor enzyme and carrier, chicken gizzard is digested, product albumen degree of hydrolysis and nitrogen recovery are higher, amino acid molecular amount and bitter taste amino acid are few, palatability is strong, has very high nutritive value.Enzymolysis product is further prepared as chicken gizzard albumen powder through precipitating, being freeze-dried, and protein content is high, substitutes Partial Fish Meal feeding animals, and growth of animal performance can be improved, and improves culture efficiency, saves aquaculture cost.
Description
Technical field
The invention belongs to complex enzyme formulation fields, and in particular to it is a kind of enzymolysis chicken gizzard complex enzyme formulation and its application.
Background technology
Chicken yield in China's grows steadily in recent years, and 2013 the 2nd, the worlds gross annual output Liang Wenju become consumption per head only
Inferior to the second largest meat products class of pork, and the 2%~2.5% of every plumage chicken is accounted for the by-product chicken gizzard of chicken association, annual
Yield reaches more than 200,000 tons.Chicken gizzard nutritive value is high, containing protein, fat, carbohydrate, vitamin A, vitamin D,
The ingredients such as phosphorus, wherein iron-holder are abundant, can play the effect of enriching blood.In addition, in chicken gizzard vitamin A content be significantly larger than milk,
The general foods such as egg, meat, fish, have the function of maintaining normal growth and reproduction, at the same can protect eyes, maintain twenty-twenty vision,
Certain popularization and application has been obtained in feed cultivation, food, medicinal etc..In existing chicken gizzard treatment process, take more
It is simple to be crushed, dry and chicken meal is made, in feed breeding production.But because this simple process easily causes nutrition
Loss, product fishy smell weight and cholesterol level height, animal palatability is poor, causes digestibility low, and apparent consumption is little, makes
At waste.Therefore, research and development improve chicken gizzard protein hydrolysis degree and utilization rate using new technique becomes hot spot of people's attention
One of problem.
Complex enzyme formulation enzymatic isolation method, which extracts albumen in chicken gizzard, becomes one of the effective way for solving chicken gizzard comprehensive utilization.It grinds
Study carefully report, by adding appropriate enzyme preparation in chicken gizzard, chicken gizzard is digested under the conditions of preference temperature, pH, reaction time, with albumen
Degree of hydrolysis is measurement index, and hydrolysate is mainly amino acid, micromolecule polypeptide and flavour nucleotide etc., have protein content and
The high feature of nutritive value can be used as by being further processed into high protein feed raw material, be added in animal and fowl fodder, recycling
It utilizes, improves chicken gizzard added value.Relatively lay particular emphasis on enzymolysis process technique study to chicken gizzard at present, as additive capacity, temperature, pH,
Extraction time etc., but enzyme preparation enzyme activity, optimum proportioning and bitter taste amino acid content can not be quantified, take which kind of enzyme preparation,
Which kind of enzyme activity, optimum proportioning enzymolysis chicken gizzard achieve the purpose that scale hydrolyzes chicken gizzard, while protein hydrolysis degree, nitrogen recycling can be improved
Rate, and reduce the generation of delicious amino acid.Chicken gizzard after enzymolysis can be by being further processed into feeding animals after protein feed, can
Reach and improve growth of animal performance, such as daily gain, daily ingestion amount, reduces feed-weight ratio, improve culture efficiency, reduce aquaculture cost.
Therefore, a kind of complex enzyme formulation that can improve chicken gizzard utilization rate of research and development is of great significance.
Invention content
In order to overcome in the prior art chicken gizzard utilization rate it is not high, the high fishy smell weight of enzymolysis product bitter taste amino acid content can not
The shortcomings that scale utilizes and deficiency, the primary purpose of the present invention is that providing a kind of complex enzyme formulation of enzymolysis chicken gizzard.
Another object of the present invention is to provide the applications of the complex enzyme formulation of above-mentioned enzymolysis chicken gizzard.
Another object of the present invention is to provide a kind of high protein feeds.
It is still another object of the present invention to provide the preparation methods of above-mentioned high protein feed.
The purpose of the invention is achieved by the following technical solution：
A kind of complex enzyme formulation of enzymolysis chicken gizzard, includes the component of following mass parts meter：
1~3 part of neutral proteinase, 1~6 part of alkali protease, 1~3 part of lipase, 0.1~0.3 part of visceral enzym, flavor
0.1~0.4 part of enzyme, 3~5 parts of carrier.
Preferably, include the component of following mass parts meter：
1~3 part of neutral proteinase, 2~6 parts of alkali protease, 1~3 part of lipase, 0.1~0.3 part of visceral enzym, flavor
0.1~0.25 part of enzyme, 3~5 parts of carrier.
It is further preferred that including the component of following mass parts meter：
1 part of neutral proteinase, 2~6 parts of alkali protease, 1 part of lipase, 0.1 part of visceral enzym, food flavor enzyme 0.1~0.25
Part, 4 parts of carrier.
Still more preferably, include the component of following mass parts meter：
1 part of neutral proteinase, 4 parts of alkali protease, 1 part of lipase, 0.1 part of visceral enzym, 0.15~0.25 part of food flavor enzyme,
4 parts of carrier.
Most preferably, include the component of following mass parts meter：
1 part of neutral proteinase, 4 parts of alkali protease, 1 part of lipase, 0.1 part of visceral enzym, 0.2 part of food flavor enzyme, carrier 4
Part.
In the complex enzyme formulation, the U/g of 50,000 U/g≤neutral proteinase enzyme activity≤100,000；100000 U/g≤basic protein
Enzyme enzyme activity≤20U/g；50000 U/g≤lipase activity≤10U/g；The U/g of 5000U/g≤visceral enzym enzyme activity≤10,000；5000U/g≤
The U/g of food flavor enzyme enzyme activity≤10,000.
The complex enzyme formulation granular size：80 mesh percent of pass >=99.0%.
The carrier includes：One or more of cornstarch, rice bran, talcum powder, wheat bran, sizing.
The application in high protein feed is made in the complex enzyme formulation of the enzymolysis chicken gizzard after digesting chicken gizzard.
A kind of high protein feed adds to be freeze-dried in chicken gizzard obtaining after above-mentioned complex enzyme formulation digests.
The preparation method of the high protein feed, includes the following steps：
(1) appropriate chicken gizzard is taken, pureed is then blended into, is uniformly mixed；
(2) 1.00~2.50% additive amount of mass percent for pressing chicken gizzard dry weight, weighs the complex enzyme of above-mentioned enzymolysis chicken gizzard
Preparation is dissolved in water, stirring to thorough dissolving；
(3) and then by complex enzyme formulation solution it is added to being preheated in 45~55 DEG C of chicken gizzard, reaction is mixed, obtains
Enzymolysis liquid；Wherein, in reaction condition, pH is 7.5~8.5, and hydrolysis temperature is 45~55 DEG C, raw material：Water=(1:1)~(6:1),
Enzymolysis time is 1.5~2.5 hours.
(4) chicken gizzard albumen powder is made after precipitating, being freeze-dried in the enzymolysis liquid that step (3) obtains；I.e.：High protein feed.
Preferably, the additive amount of the complex enzyme formulation of the enzymolysis chicken gizzard described in step (2) is 1.50~2.50%；It is more excellent
It is selected as 1.50~2.00%；More preferably 1.50%.
Preferably, the reaction condition described in step (4) is：PH is 8.0, and hydrolysis temperature is 50 DEG C, raw material：Water=4：1,
Enzymolysis time is 2.0 hours.
Above-mentioned preparation method increases product albumen degree of hydrolysis and nitrogen recovery.
Above-mentioned preparation method reduces enzymolysis product delicious amino acid.
Application of the high protein feed in substituting Partial Fish Meal for feeding animals.
Above-mentioned high protein feed is substituted into Partial Fish Meal feeding animals, growth of animal performance can be improved, improves chicken gizzard quality
And its utilization rate in actual production, high-quality protein source is provided for feed cultivation, promotes food security.
The present invention has the following advantages and effects with respect to the prior art：
The present invention by neutral proteinase, alkali protease, lipase, visceral enzym, food flavor enzyme and carrier by different enzyme activity,
Proportioning, dosage compounding are complex enzyme formulation, are digested to chicken gizzard, product albumen degree of hydrolysis and nitrogen recovery are higher, amino acid
Molecular weight and bitter taste amino acid are few, and palatability is strong, have very high nutritive value.Enzymolysis product through precipitate, be freeze-dried into
One step is prepared as chicken gizzard albumen powder, and protein content is high, substitutes Partial Fish Meal feeding animals, growth of animal performance can be improved, and improves
Culture efficiency saves aquaculture cost.
Description of the drawings
Fig. 1 is influence of the different composite enzyme preparation to chicken gizzard protein hydrolysis degree in embodiment 1.
Fig. 2 is influence of the different composite enzyme preparation to chicken gizzard nitrogen recovery in embodiment 1.
Fig. 3 is the different composite enzyme preparation influence to chicken gizzard protein hydrolysis degree under given conditions in embodiment 2.
Fig. 4 is molecular weight of albumen testing result in enzymolysis liquid after different composite enzyme preparation enzymolysis in embodiment 2.
Fig. 5 is influence of the various dose complex enzyme formulation 3 to chicken gizzard protein hydrolysis degree in embodiment 3.
Fig. 6 is influence of the various dose complex enzyme formulation 3 to chicken gizzard nitrogen recovery in embodiment 3.
Specific implementation mode
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Neutral proteinase used in embodiment, alkali protease, lipase are that the green micro- health Animal nutrition in Shenzhen is limited
Company produces, and universal, model is respectively ZD500, JD1000, F800.Visceral enzym derives from pluck, buys in Nan Ningdong
The roads Heng Hua bio tech ltd, model NZ100.Food flavor enzyme is bought in Pangbo Bioengineering Co Ltd, Nanning, model
For FW100.
Carrier is bought for COFCO Biochemical Energy (Yushu) Co., Ltd..
Embodiment 1
1, in complex enzyme formulation each enzyme activity need to meet it is claimed below：
The U/g of 50000 U/g≤neutral proteinase enzyme activity≤100,000；100000 U/g≤alkali protease enzyme activity≤20U/g；50000 U/g
≤ lipase activity≤10U/g；The U/g of 5000U/g≤visceral enzym enzyme activity≤10,000；The U/g of 5000U/g≤food flavor enzyme enzyme activity≤10,000.
2, complex enzyme formulation granular size：80 mesh percent of pass >=99.0%.
3, the carrier includes：One or more of cornstarch, rice bran, talcum powder, wheat bran, sizing.
4, lipase, visceral enzym, food flavor enzyme and carrier number are fixed in complex enzyme formulation, neutral proteinase and basic protein
Enzyme is complex enzyme formulation according to different proportion proportioning.
(1) complex enzyme formulation 1：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：2：1：0.1：0.2：4；
(2) complex enzyme formulation 2：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：3：1：0.1：0.2：4；
(3) complex enzyme formulation 3：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.1：0.2：4；
(4) complex enzyme formulation 4：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：5：1：0.1：0.2：4；
(5) complex enzyme formulation 5：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：6：1：0.1：0.2：4.
5, it takes freezing chicken gizzard several, thaws and then blend into pureed, be uniformly mixed.
(1) 3 250mL conical flasks are taken, 200g minced chicken liver is weighed respectively and is placed in conical flask, it is entirely warm to place HZQ-F160Q
55 DEG C of concussion and cultivate case is spent, rate 220r/min is shaken, preheats 20min.
(2) 1.50% additive amount is pressed, different composite enzyme preparation 1,2,3,4,5 is weighed, is placed in 50mL beakers, 20mL is added
Water is placed on HJ-4 magnetic force heating stirrers stirring 10min, enzyme is made thoroughly to be dissolved in water.
(3) 5 groups of complex enzyme formulation solution are added in the conical flask of the preheating equipped with chicken gizzard respectively, are mixed.Continue
55 DEG C of HZQ-F160Q total temperatures concussion and cultivate case is placed, rate 220r/min is shaken, is reacted 2.5 hours.
(4) it after the completion of reacting, places 5~10 minutes and inactivates in 100 DEG C of water-baths.
(5) after the completion of inactivating, with desk centrifuge 5000r/min, centrifugation reaction 15 minutes.
(6) graduated cylinder for being 100mL with range measures 3 groups of supernatant volumes.
(7) nitrogen content in supernatant and chicken gizzard raw material is surveyed with Kjeldahl's method.
(8) content that amino nitrogen in its 3 groups of supernatants is surveyed with formaldehyde potentiometric titration, is multiplied by supernatant volume and obtains
The quality of amino nitrogen in clear liquid.
(9) molecular weight of albumen size distribution in supernatant is surveyed with gel electrophoresis.
6, albumen in complex enzyme formulation enzymolysis chicken gizzard, additive capacity is that 1.00~2.00%, pH is 7.5~8.5, enzymolysis temperature
Degree is 45~55 DEG C, raw material：Water=(1:1)~(6:1), enzymolysis time is 1.5~2.5 hours, show that additive capacity is
1.5%, optimum pH are 8.0, and hydrolysis temperature is 50 DEG C, raw material：Water=4：1, enzymolysis time is 2 hours.
7, testing index：Amino nitrogen, protein hydrolysis degree, nitrogen recovery.
8, testing index：
The measurement of amino-acid nitrogen content refers to formaldehyde potentiometric titration；
Nitrogen pool, which measures, refers to Kjeldahl's method；
The assay method of protein hydrolysis degree in enzymolysis liquid：
Protein hydrolysis degree (DH, %)=(total nitrogen quality in amino nitrogen quality/sample in supernatant) × 100%；
Nitrogen recovery (NR, %)=(total nitrogen content in amino-acid nitrogen content/raw material in supernatant) × 100%.
9, hydrolysis result of the different composite enzyme preparation to chicken gizzard
Complex enzyme formulation 1~5 is under given conditions to the enzymolysis of chicken gizzard as a result, as shown in table 1 and Fig. 1,2.
1 different composite enzyme preparation of table is under given conditions to the enzymolysis result of chicken gizzard
| Different enzyme preparations | Protein hydrolysis degree DH (%) | Nitrogen recovery DR (%) |
| Complex enzyme formulation 1 | 28.20 | 94.83 |
| Complex enzyme formulation 2 | 29.29 | 97.18 |
| Complex enzyme formulation 3 | 31.55 | 99.71 |
| Complex enzyme formulation 4 | 30.03 | 96.81 |
| Complex enzyme formulation 5 | 28.92 | 95.26 |
By table 1, Fig. 1 and Fig. 2 it is found that in additive capacity 1.5%, pH 8.0, hydrolysis temperature is 50 DEG C, raw material：Water=4：
1, enzymolysis time is 2 hours, and the hydrolysis result of complex enzyme formulation 3 is best, i.e. neutral proteinase：Alkali protease：Lipase：It is interior
Dirty enzyme：Food flavor enzyme：Carrier=1：4：1：0.10：0.10：It is higher to the protein hydrolysis degree of chicken gizzard, nitrogen recovery is high under 4 proportionings.
Analyze possible reason：Work as neutral proteinase：Alkali protease：Lipase is according to 1：4：Under 1 proportioning, it is in pH
Under 8.0 environment, it is in complex enzyme formulation optimal pH state, the decomposition rate of complex enzyme formulation can be played to greatest extent, by chicken gizzard
In protein breakdown be amino acid and micromolecule polypeptide, so complex enzyme formulation 3 is best to the hydrolysis result of chicken gizzard, proteolysis
Degree and nitrogen recovery highest.
Embodiment 2
1, neutral proteinase, alkali protease, lipase, visceral enzym, carrier number are constant, change food flavor enzyme number, survey
Determine the hydrolysis result of chicken gizzard, protein molecular weight and delicious amino acid content.
(1) complex enzyme formulation 6：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.10：0.05：4；
(2) complex enzyme formulation 7：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.10：0.10：4；
(3) complex enzyme formulation 8：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.10：0.15：4；
(4) complex enzyme formulation 9：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.10：0.20：4；
(5) complex enzyme formulation 10：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.10：0.25：4.
2, complex enzyme formulation enzymolysis chicken gizzard, additive capacity 1.0~2.0%, pH 7.5~8.5,45~55 DEG C of hydrolysis temperature,
Raw material：Water=(1:1)~(6:1), enzymolysis time 1.5~2.5 hours obtain additive capacity 1.50%, optimum pH8.0, enzyme
Solve temperature 50 C, raw material：Water=4：1, it is 2 hours when enzymolysis.
3, testing index：Nitrogen pool, amino nitrogen, protein hydrolysis degree, protein molecular weight, free amino acid composition contain
Amount.
4, assay method：
The measurement of amino-acid nitrogen content refers to formaldehyde potentiometric titration；
Nitrogen pool, which measures, refers to Kjeldahl's method；
The assay method of protein hydrolysis degree in enzymolysis liquid：
Protein hydrolysis degree (DH, %)=(total nitrogen quality in amino nitrogen quality/sample in supernatant) × 100%；
Protein molecular weight refers to SDS-PAGE electrophoresis technique determining protein Measuring Molecule Weights.
Free amino acid forms the measurement of content：Using automatic amino acid analyzer analyze 18 kinds of free amino acids composition and
Content.It accurately takes sample 3mL in 5mL plastic centrifuge tubes, the concussion of 42% sulfosalisylic acid solutions of 0.5mL is added and shakes up.Refrigerator
2min is centrifuged in 13000r/min after standing 12 hours, is analyzed with machine on 0.22m membrane filtrations.
, there are 2 columns, splitter in each sample analysis period 53min, when analysis：4.6mm × 60mm, eluent flow rate
0.4mL/min, 700 DEG C of column temperature, column press 11.627MPa；Reaction column：Ninhydrin and ninhydrin buffer flow rate 0.35mL/min,
135 DEG C of column temperature, column press 1.078MPa.
E/T values=essential amino acid/total amino acid content (essential amino acid：Lysine (Lys), phenylalanine (Phe), figured silk fabrics
Propylhomoserin (Val), leucine (Leu), threonine (Thr), isoleucine (Ile), methionine (Met), tryptophan (Trp))
TAV values (flavour activity value) are the ratio of certain amino acid composition concentration and its threshold value.
5, influence of the different composite enzyme preparation to chicken gizzard protein hydrolysis degree
Complex enzyme formulation 6~10 is in the influence under given conditions to chicken gizzard protein hydrolysis degree, as shown in table 2 and Fig. 3.
The influence to chicken gizzard protein hydrolysis degree under given conditions of 2 different composite enzyme preparation of table
| Different composite enzyme preparation | Protein hydrolysis degree DH (%) |
| Complex enzyme formulation 6 | 29.72 |
| Complex enzyme formulation 7 | 30.08 |
| Complex enzyme formulation 8 | 30.63 |
| Complex enzyme formulation 9 | 31.55 |
| Complex enzyme formulation 10 | 30.96 |
By table 2, Fig. 3 it is found that complex enzyme formulation 9 is to the protein hydrolysis degree highest of chicken gizzard, hydrolysis effect is best.Analysis may
The reason of be：9 neutral proteinase of complex enzyme formulation：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier proportioning is 1：4：
1：0.10：0.20：4, it is in optimum proportioning and optimal pH, the enzymatic catalysis of the various enzyme preparations in complex enzyme formulation reaches most
Good discomposing effect.
6, protein molecular weight detects after different composite enzyme preparation enzymolysis chicken gizzard
Molecular weight of albumen testing result in enzymolysis liquid after complex enzyme formulation 6~10 digests, as shown in Figure 4.
Fig. 4, supernatant of the chicken gizzard after the enzymolysis of different composite enzyme preparation 6,7,8,9,10 use SDS- after 10 times of dilution
Protein molecular weight in PAGE electrophoresis technique determining supernatants.As can be known from Fig. 4, the main production after 5 kinds of complex enzyme formulation enzymolysis
Object be molecular weight between 14.4~20KD, the small peptide of 20~27KD, but have in 9 product of complex enzyme formulation a small amount of molecular weight between
The small peptide of 9.5~14.4KD, this illustrates that chicken gizzard is more thorough after 9 enzymolysis of complex enzyme formulation, and effect is more apparent.
7, free amino acid forms content in enzyme solution after different composite enzyme preparation digests chicken gizzard
Free amino acid composition content (g/dL) in enzyme solution after 6~10 pairs of chicken gizzard enzymolysis of complex enzyme formulation, as shown in table 3.
Free amino acid composition content (g/dL) in enzyme solution after 3 different composite enzyme preparation of table digests chicken gizzard
As shown in Table 3, chicken gizzard is after the enzymolysis of different composite enzyme preparation 6,7,8,9,10, essential amino acid in enzyme solution (Lys,
Phe, Val, Leu, Trp, Thr, Ile, Met) account for the ratio (E/T) of free amino acid be respectively 49.37%, 49.88%,
49.25%, 49.07%, 49.70%, difference is little, and ratio is above WHO/FAO standards (35.38%).Essential amino acid profit
In digesting and assimilating for livestock and poultry, content is higher, and nutritive value is higher, is added in feed and is more easy to be absorbed by animal body.
8, delicious amino acid accounts for the content of free amino acid after different composite enzyme preparation enzymolysis chicken gizzard
Delicious amino acid accounts for the content of free amino acid after the enzymolysis chicken gizzard of complex enzyme formulation 6~10, as shown in table 4.
4 delicious amino acid of table accounts for the ratio of free amino acid
As shown in Table 4, after different composite enzyme preparation 6,7,8,9,10 digests chicken gizzard, aspartic acid (Asp), paddy ammonia in product
The sum of sour (Glu) and proline (Pro) this 3 kinds of Fresh ear field contents are in rising trend, wherein chicken gizzard is through complex enzyme formulation 9
After enzymolysis, product Fresh ear field content highest, effect is most apparent.Threonine (Thr), serine (Ser), glycine (Gly),
The sum of alanine (Ala) this 4 kinds of sweet taste amino acid content is not much different.Sweet tea/bitter taste amino acid valine (Val), methionine
(Met), the sum of lysine (Lys), arginine (Arg) content are in the trend being gradually reduced；Bitter taste amino acid content is also in gradual
Downward trend, wherein 9 content of complex enzyme formulation is minimum, and effect is best.
Chicken gizzard is after different composite enzyme preparation enzymolysis, by the content of 4 kinds of delicious amino acids in table 4 it is found that enzymolysis product contains
It is little to measure difference, but 9 effect of complex enzyme formulation is relatively preferable.
Embodiment 3
1, complex enzyme formulation 3：
Neutral proteinase：Alkali protease：Lipase：Visceral enzym：Food flavor enzyme：Carrier=1：4：1：0.1：0.2：4.
2, additive capacity, if 5 gradients：0.50%, 1.00%, 1.50%, 2.00%, 2.50%；
3, using chicken gizzard as raw material, complex enzyme formulation 3 is in optimum pH 8.0,50 DEG C of hydrolysis temperature, raw material：Water=4：1, enzyme
Solve time 2 h.
4, testing index：Total protein nitrogen, amino nitrogen, protein hydrolysis degree, nitrogen recovery.
5, assay method：
Total protein nitrogen：Kjeldahl's method；
Amino nitrogen：Formaldehyde potentiometric titration；
Protein hydrolysis degree (DH, %)=(total nitrogen quality in amino nitrogen quality/sample in supernatant) × 100%；
Nitrogen recovery (NR, %)=(total nitrogen content in amino-acid nitrogen content/raw material in supernatant) × 100%.
6, hydrolysis result of the different graded composite enzyme preparations 3 to chicken gizzard
Various dose complex enzyme formulation 3 is under given conditions to the enzymolysis of chicken gizzard as a result, as shown in table 5 and Fig. 5,6.
5 various dose complex enzyme formulation 3 of table is under given conditions to the enzymolysis result of chicken gizzard
| Additive capacity (%) | Protein hydrolysis degree DH (%) | Nitrogen recovery DR (%) |
| 0.50 | 26.63 | 93.67 |
| 1.00 | 29.06 | 95.83 |
| 1.50 | 31.55 | 99.71 |
| 2.00 | 30.74 | 98.64 |
| 2.50 | 30.07 | 98.02 |
By table 5, Fig. 5, Fig. 6 it is found that when additive capacity is 1.50%, protein hydrolysis degree of the complex enzyme formulation 3 to chicken gizzard
Highest, while nitrogen recovery also highest.Reaction is initial, and as additive capacity gradually increases, enzyme-to-substrate contacts density and increases, enzyme
Promote reaction rate to accelerate, discomposing effect is good, and protein hydrolysis degree and nitrogen recovery are high.When additive capacity is up to 1.50%, albumen water
Xie Du and nitrogen recovery have reached highest, if being further continued for adding, protein hydrolysis degree and nitrogen recovery are without too big variation.
Embodiment 4
The enzymolysis liquid of complex enzyme formulation 6,7,8,9,10 through precipitating, be freeze-dried, be respectively prepared protein feed 1,2,3,4,
5,2% fish meal feeding suckling piglet is substituted respectively.
(1) experiment chooses that 288 weight are close, the good 21 age in days weanling pig (Ternary Pig) of upgrowth situation, random point
For 6 groups, 4 repetitions are each organized, each repeat 12 pigs, the 1st group is positive control group, and the 2nd, 3,4,5,6 group is respectively albumen
Feed 1,2,3,4,5 groups substitute 2% fish meal respectively.
(2) diet recipe and trophic level
It tests Diet Formula to design according to U.S. NRC (2012) pig nutritional requirements, using corn-soybean meal basic day
Grain is shown in Table 6.
6 basal diet formula of table and trophic level
| Raw material | Proportioning | Trophic level * * | Content |
| Corn (%) | 49.87 | Digestible energy (MJ/kg) | 14.23 |
| Soybean Meal (%) | 34.16 | Thick protein (%) | 23.70 |
| Expanded soybean (%) | 4.91 | Crude fat (%) | 3.51 |
| Whey powder (%) | 2.68 | Calcium (%) | 0.80 |
| Soy protein concentrate (%) | 2.17 | Total phosphorus (%) | 0.65 |
| Fish meal (%) | 2.17 | Available phosphorus (%) | 0.41 |
| Soya-bean oil (%) | 1.07 | L-lysine (%) | 1.36 |
| Calcium monohydrogen phosphate (%) | 0.84 | Egg+cystine (%) | 0.76 |
| Mountain flour (%) | 0.83 | ||
| Salt (%) | 0.30 | ||
| Additive premix * (%) | 1.00 | ||
| It is total | 100.00 |
Note：* premix provides vitamin A 8-15 (KIU), vitamine D3 (KIU) 1.5-3.0, dimension life for every kilogram of diet
Plain E (mg) >=32, Vitamin K3 (mg) >=2.5, iron (mg) >=175, copper (mg) 125-200, zinc (mg) 200-2250, manganese (mg)
≥75；* trophic level is calculated value, dry matter content 88%.
(3) feeding management
On the Guangzhou Huadu District towns the Hua Dong pig farms Xin Tian, experimental period is 28 days for test site.Pig house is semi-enclosed, high bed
Slatted floor.Experimental period piglet is freely eaten, and immune and other management are operated according to the normal production management program in pig farm.
(4) testing index and method
1, average daily gain (ADG)：
Average daily gain (ADG)=(the last weight-experiment initial weight of experiment)/experiment number of days
2, average daily gain (ADFI)：
Average daily gain (ADFI)=(the total clout amount of total amount of feeding -)/test period
3, feed consumption weightening is than (FCR)：
Feed consumption weightening ratio=average daily gain (ADFI)/average daily gain (ADG)
4, diarrhea rate (DR)：
Diarrhea rate=experimental period diarrhea piglet head/(experiment piglet head number * tests number of days) * 100%
(5) measurement result
The different protein feed groups of table 7 substitute influence of 2% fish meal to suckling piglet growth performance
Compare with control group, protein feed 2,3,4,5 groups of daily gains increase separately 7.4%, 10.4%, 12.5%,
10.2%, equal significant difference (P ＜ 0.05)；In terms of daily ingestion amount, protein feed 1,2,3,4,5 groups increase separately 1.6%,
4.8%, 5.4%, 6.1%, 5.3%, equal difference is not notable (P ＞ 0.05)；Feed consumption increases weight in terms of ratio, protein feed 3,4,5 groups
4.8%, 5.9%, 4.3% is reduced respectively, equal significant difference (P ＜ 0.05).In terms of diarrhea rate, protein feed 1,2,3,4,5 groups
20.1%, 29.2%, 39.6%, 36.1%, 33.3% is reduced respectively, equal significant difference (P ＜ 0.05).
As known from Table 7, with protein feed 3,4,5 groups of 2% fish meal of replacement, suckling piglet daily gain can be effectively improved and reduce material
Compare again, improves growth performance, while diarrhea rate is substantially reduced.
Embodiment 5
(1) experimental animal grouping and processing
Select 48 ages in days, weight about 15kg (weight differences are not notable), growth body condition good, Du of castration × (length
× big) child care pig, it is randomly divided into 6 groups, control group, protein feed 1,2,3,4,5 groups, every group of 4 repetitions each repeat 2 pigs.
(2) diet recipe and trophic level, with embodiment 4.
(3) feeding management
Experiment carries out in the buildings Agricultural University Of South China animal science institute Wei Nan, takes single cage digestion trial device to raise, pig
It is 25~30 DEG C to give up temperature control, natural lighting, and routinely operating instruction carries out for feeding management.
(4) test method
This digestion trial receives excrement method using complete.Daily early 8：00 point and evening 6：00 point quantitatively feeds intake, one hour recession material
Slot, and clout weight in hopper is weighed, change sink.Concrete operations are as follows：
Laundering period (the 1st~7 day) feeds complete diet pellet, is freely eaten, records every pig daily feed intake.
Pre-feeding period (the 8th~12 day), by 85% feeding experiment material of every pig feed intake.
Positive to raise the phase (the 13rd~16 day), by 85% feeding experiment material of every pig voluntary feed intake, just raising, the 1st day phase is early
Start within 1 hour to receive excrement after morning feeding, continuously feed and receive excrement 4 days, i.e., to 1 hour stopping receipts excrement after early feeding in the 17th day；Receive excrement
After spray a small amount of 10% hydrochloric acid, while recording the daily inventory and clout amount of every pig, total scale of feeding subtracts remaining in the positive feeding phase
Doses is feed intake.
As unit of repetition, excrement sample collected by positive 4 days feeding phases is placed in 65 DEG C of baking ovens and is dried to constant weight, is got damp again at room temperature
It 24 hours, weighs, then crushed 40 mesh sieve, air-drying sample is made and is that 4 DEG C of refrigerators save backup.
(5) digestion trial testing index and method
Dry matter (DM) is measured with reference to the method for GB/T6435-2006 in daily ration and excrement sample；Crude protein (CP) is with reference to GB/
The method of T6432-1994 carries out pre-treatment, is then measured using FOSS Kjeltec-2300 full-automatic Kjeldahl determination devices；
Crude fat (EE) is measured with reference to GB/T6433-2006 using Shanghai fibre inspection SZC-C type Milko-Testers；Total energy (GE) makes
It is measured with IKAC2000 calorimeters.
(6) nutritional ingredient digestibility computational methods
(7) Biochemical Indices In Serum measurement and method
Serum sample acquisition carries out after digestion trial.Start feeding within the 1st day after digestion trial to be freely eaten.
Morning 8 on day 4：Before 00 feeding, vena cava anterior blood sampling 20mL is carried out to every pig, room temperature shady place, which tilts, to be stood, blood to be had
Clear 3000rpm is centrifuged 10 minutes, and upper serum is taken to dispense when being precipitated, and sets that put -80 DEG C of preservations after ice chest precooling to be measured, using examination
Agent box (being built up purchased from Nanjing) detect serum high-density LP (HDL), low-density lipoprotein (LDL), total cholesterol (TC) and
Free fatty (NEFA), and calculate high-density lipoprotein and ldl ratio.
(8) test result
The different protein feed groups of table 8 substitute influence (%) of 2% fish meal to suckling piglet nutritional ingredient digestibility
As shown in Table 8, compared with the control group, protein feed 1, the dry matter in 2,3,4,5 groups, protein, crude fat and
Total energy apparent digestibility is all improved to some extent, but difference is not notable (P ＞ 0.05).Compared with the control group, protein feed
2,3,4,5 histone matter digestibilities are respectively increased 3.5%, 3.9%, 4.3%, 3.6%, equal significant difference (P ＜ 0.05).
9 protein feed group of table substitutes influence of 2% fish meal to suckling piglet Biochemical Indices In Serum
As shown in Table 9, free fatty, compared with the control group, protein feed 2,3,4,5 groups reduce by 12.9% respectively,
15.0%, 25.8%, 19.1%, equal significant difference (P ＜ 0.05)；In terms of total cholesterol, albumen 3,4,5 groups reduce respectively
22.2%, 27.8%, 24.1%, equal significant difference (P ＜ 0.05)；In terms of high-density lipoprotein, compared with the control group, albumen is raised
3,4,5 groups of material increases 18.4%, 25.0%, 21.1% respectively, equal significant difference (P ＜ 0.05)；In terms of low-density lipoprotein, with
Control group compares, and 4,5 groups of protein feed reduces by 33.0%, 27.1% respectively, equal significant difference (P ＜ 0.05)；The side HDL/LDL
Face, protein feed 3,4,5 groups of raisings 48.3%, 87.6%, 66.3%, equal significant difference (P ＜ 0.05).
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications,
Equivalent substitute mode is should be, is included within the scope of the present invention.
Claims (10)
1. a kind of complex enzyme formulation of enzymolysis chicken gizzard, it is characterised in that include the component of following mass parts meter：
1~3 part of neutral proteinase, 1~6 part of alkali protease, 1~3 part of lipase, 0.1~0.3 part of visceral enzym, food flavor enzyme 0.1
~0.4 part, 3~5 parts of carrier.
2. the complex enzyme formulation of enzymolysis chicken gizzard according to claim 1, it is characterised in that include the group of following mass parts meter
Point：
1~3 part of neutral proteinase, 2~6 parts of alkali protease, 1~3 part of lipase, 0.1~0.3 part of visceral enzym, food flavor enzyme 0.1
~0.25 part, 3~5 parts of carrier.
3. the complex enzyme formulation of enzymolysis chicken gizzard according to claim 2, it is characterised in that include the group of following mass parts meter
Point：
1 part of neutral proteinase, 2~6 parts of alkali protease, 1 part of lipase, 0.1 part of visceral enzym, 0.1~0.25 part of food flavor enzyme carry
4 parts of body.
4. the complex enzyme formulation of enzymolysis chicken gizzard according to claim 3, it is characterised in that include the group of following mass parts meter
Point：
1 part of neutral proteinase, 4 parts of alkali protease, 1 part of lipase, 0.1 part of visceral enzym, 0.15~0.25 part of food flavor enzyme, carrier
4 parts.
5. digesting the complex enzyme formulation of chicken gizzard according to Claims 1 to 4 any one of them, it is characterised in that：
In the complex enzyme formulation, the U/g of 50,000 U/g≤neutral proteinase enzyme activity≤100,000；100000 U/g≤alkali protease enzyme
Work≤20U/g；50000 U/g≤lipase activity≤10U/g；The U/g of 5000U/g≤visceral enzym enzyme activity≤10,000；5000U/g≤flavor
The U/g of enzyme enzyme activity≤10,000；
The complex enzyme formulation granular size：80 mesh percent of pass >=99.0%；
The carrier includes：One or more of cornstarch, rice bran, talcum powder, wheat bran, sizing.
6. high protein feed is made after digesting chicken gizzard in the complex enzyme formulation of Claims 1 to 5 any one of them enzymolysis chicken gizzard
In application.
7. a kind of high protein feed, it is characterised in that：It is that Claims 1 to 5 any one of them enzymolysis chicken is added in chicken gizzard
It is freeze-dried and obtains after the complex enzyme formulation enzymolysis of liver.
8. the preparation method of the high protein feed described in claim 7, it is characterised in that include the following steps：
(1) appropriate chicken gizzard is taken, pureed is then blended into, is uniformly mixed；
(2) 1.00~2.50% additive amount of mass percent for pressing chicken gizzard dry weight, weighs the complex enzyme formulation of above-mentioned enzymolysis chicken gizzard
It is dissolved in water, stirring to thorough dissolving；
(3) and then by complex enzyme formulation solution it is added to being preheated in 45~55 DEG C of chicken gizzard, reaction is mixed, is digested
Liquid；Wherein, in reaction condition, pH is 7.5~8.5, and hydrolysis temperature is 45~55 DEG C, raw material：Water=(1:1)~(6:1) it, digests
Time is 1.5~2.5 hours；
(4) chicken gizzard albumen powder is made after precipitating, being freeze-dried in the enzymolysis liquid that step (3) obtains；I.e.：High protein feed.
9. the preparation method of high protein feed according to claim 8, it is characterised in that：
The additive amount of the complex enzyme formulation of enzymolysis chicken gizzard described in step (2) is 1.50~2.50%；
Reaction condition described in step (4) is：PH is 8.0, and hydrolysis temperature is 50 DEG C, raw material：Water=4：1, enzymolysis time is
2.0 hour.
10. application of the high protein feed in substituting Partial Fish Meal for feeding animals described in claim 7.
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| CN109371089A (en) * | 2018-12-25 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of extracting method of small molecule liver peptide |
| CN110292109A (en) * | 2019-06-06 | 2019-10-01 | 光泽县泽农生物科技有限公司 | A method of improving chicken liver meal utilization rate |
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| CN109371089A (en) * | 2018-12-25 | 2019-02-22 | 河北肽都生物科技有限公司 | A kind of extracting method of small molecule liver peptide |
| CN110292109A (en) * | 2019-06-06 | 2019-10-01 | 光泽县泽农生物科技有限公司 | A method of improving chicken liver meal utilization rate |
| CN110915995A (en) * | 2019-12-03 | 2020-03-27 | 江苏省农业科学院 | Method for preparing pet feed additive from poultry liver protein hydrolysate |
| CN112617025A (en) * | 2020-12-11 | 2021-04-09 | 桐城市雨润生物科技有限公司 | Preparation method and application of animal viscera powder |
| CN116250614A (en) * | 2021-12-08 | 2023-06-13 | 南京钦润生物科技有限公司 | Solid preparation method of chicken liver powder |
| CN116019178A (en) * | 2022-12-23 | 2023-04-28 | 安佑生物科技集团股份有限公司 | Creep feed for weaned pigs and preparation method thereof |
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