CN101530162A - Preparation method of novel protein feed additive - Google Patents

Abstract

The invention relates to a preparation method of a hydrolyzed protein mixture and a protein peptide powder feed additive. The method uses a multi-enzyme compound enzymatic method to hydrolyze plant protein to prepare the hydrolyzed protein mixture and protein peptide powder. Compounding can increase the speed and degree of protein hydrolysis, improve feed utilization, and reduce production costs. The product prepared by the preparation method can be used as a feed additive to significantly increase the egg production rate of laying poultry; increase the weight gain of broiler poultry, reduce the feed-to-meat ratio, and improve the production performance of broiler chickens; it can also reduce the incidence of diarrhea in young animals, reduce the Feed-to-meat ratio of piglets; used in aquatic animals, it can increase the weight gain of fish, reduce the bait coefficient, and reduce the viscosity of intestinal contents.

Description

A kind of preparation method of novel protein feed additive

Technical field

The invention belongs to feed additive field, relate to a kind of preparation method of novel protein feed additive, specifically relate to the preparation method of a kind of protein hydrolysate mixture and protein peptide powder feed addictive.

Background technology

The feed resource deficiency, particularly the protein resource shortage in the feed is the subject matter of restriction China aquaculture development, and, on protein feed resource utilizes, China also exists conversion ratio and the lower problem of utilization rate, therefore, exploitation also rationally utilizes protein feed significant to the development of aquaculture.

Discover that existing protein feed often exists ANFs and toxicity problem, the security that influence is used; On the other hand, the protein major part that is digested in the feed is with oligopeptide rather than absorbed with single amino acid, and power consumption was low when various peptides were transferred in the zooblast, running is fast, and function is strong, and the utmost point is beneficial to animal digesting and assimilating protein feed.Therefore, plant albuminoid feeds such as dregs of beans being carried out predigestion, make it to become vegetalitas small peptide product, reduce ANFs content, is the effective way that obtains the high-quality forage protein.

Research to peptide starts from late 1970s in the world, and China began one's study in recent years, but prepared product is used for food and health products more.At present, the peptide product production method has both at home and abroad: acid hydrolyzation, chemical synthesis, fermentation method (solution fermentation, solid state fermentation), enzyme process.The drawback of acid hydrolyzation is to be difficult to control hydrolysis degree, destroys tryptophan and some hydroxyaminos easily, and hydrolysate is unfavorable for the oligopeptides preparation based on amino acid.Chemical synthesis is used for the pharmacology level peptide of the short chain of production high value to long-chain, and the shortcoming of this method is the cost height, and may be harmful to healthy and environment in course of reaction.Solution fermentation prepares Soybean Peptide and needs high-temperature sterilization and airtight condition, the cost height; The working condition of bacterium and enzyme there are differences, and can not satisfy both requirements of growing simultaneously; Hydrolysis degree can not accurately be controlled, unstable product quality etc.Solid state fermentation commonly used has advantages such as energy consumption is low, process is economized on water, easy to operation in feedstuff industry at present, but ventilation or heat radiation difficulty, product quality is difficult to control equally.Production by Enzymes has that production scale is big, cost is low, gentle harmless, the low little peptide yield of molecule of reaction condition is higher, technology is relatively easily controlled, be applicable to advantage such as suitability for industrialized production.Enzyme process has purposes more widely in food service industry, but is mostly to adopt single protease, and is limited to the destruction of degradation of protein and ANFs thereof, is difficult to be applied in practice.

Patent CN1530022 discloses a kind of preparation method of protein feed, and this method adopts biological fermentation process to be prepared in conjunction with enzyme process, and ANFs is removed in fermentation on the one hand, with enzyme albumen is degraded fast on the other hand; But, consider the growth cycle of bacterium, fermentation time is still longer; And bacterium and enzyme are added simultaneously, can not take into account both optimum reaction conditionses; In addition, adopt solid fermentation method, unstable product quality.Disclose the method with liquid Production by Enzymes Soybean Peptide among patent CN1377590 and the CN1274361, the two all adopts single protease, only is hydrolyzed at albumen, the SNSP in the vegetable protein is not degraded.Patent CN1957736 discloses the production method of a kind of soybean protein (peptide) powder, and this method relates to a kind of method of comprehensive development and utilization dregs of beans, but because its technology is loaded down with trivial details, is unfavorable for the application of low value-added feedstuff industry.

Summary of the invention

It is simple to the invention provides a kind of technology, is suitable for producing at the zymyhydrolyzed protein feedstuff industry suitability for industrialized production, that multienzyme is compound the method for protein hydrolysate mixture and protein peptide powder feed addictive.

The objective of the invention is to overcome the art methods complexity, it is consuming time long to ferment, and the drawback that hydrolysis is single, degree of hydrolysis is low improves hydrolysis rate and hydrolysis degree, and improves efficiency of feed utilization, reduces production costs.

The objective of the invention is to be achieved through the following technical solutions:

The invention provides a kind of preparation method of novel protein feed additive, comprise the steps:

1) pretreatment of raw material:

The enzymolysis substrate is crushed to 40～100 orders, put into enzymatic vessel, add entry, add the weight of entry and the weight proportion of the contained protein of enzymolysis substrate is 100:5～18, stir, transfer pH to 7～9, soaked 0.5～1 hour under 30～37 ℃ of conditions, then, 80～100 ℃ of down sterilizations 10～20 minutes, after cool to 50～60 ℃;

2) enzymolysis processing of complex enzyme:

In the pretreated raw material that step 1) obtains, add non-starch polysaccharide enzyme (NSP enzyme) successively, protease is hydrolyzed, 45～60 ℃ of temperature, the pH nature, enzymolysis 6～14h stopped reaction at 20～42% o'clock to degree of hydrolysis (DH%);

3) enzyme that goes out:

With step 2) liquid after the enzymolysis processing that obtains is in 90～120 ℃ of enzymes 15～30 minutes of going out, and flow out liquid and be protein enzymatic hydrolyzate;

4) concentrate:

With the protein enzymatic hydrolyzate that step 3) obtains, be concentrated into solid content 15～35%, get concentrate;

5) spray-drying:

The concentrate of step 4) is squeezed into the press spray drying tower through high-pressure pump, 170～190 ℃ of air intakes, 70～85 ℃ of air drafts, temperature is 100～110 ℃ in the tower, carries out spray-drying, obtains the protein hydrolysate mixture.

Wherein, described enzymolysis substrate is following vegetable protein: any in soybean protein isolate, FSPC, dregs of beans, soya-bean cake, cotton cake dregs, dregs of rapeseed cake, sunflower seeds grouts, the peanut dregs, two or more mixture.

Non-starch polysaccharide enzyme among this preparation method (NSP enzyme) can reduce the content of SNSP in the feed such as dregs of beans, increases the content of reduced sugar in protein hydrolysate mixture and the protein peptides, improves hydrolysis rate and hydrolysis degree.Non-starch polysaccharide enzyme can in pectase, cellulase, the zytase any, two or more; Described protease is neutral proteinase and/or alkali protease, and protease can be following bacterium: any in protease that hay bacillus 1398, actinomyces 166, aspergillus terricola 3942, aspergillus niger 3350 and/or bacillus licheniformis 2709 produce and hydrolysising protease (Alcalase), flavor protease (Flavourzyme), the compound protease (Protamex), two or more.

Further, the addition of described pectase, cellulase, zytase and protease is: pectase, 0～3000U/g albumen; Cellulase, 0～2000U/g albumen; Zytase, 0～4000U/g albumen; Protease, 1000～10000U/g albumen.

All can be used as feed addictive by prepared protein enzymatic hydrolyzate of the above step of the present invention and protein hydrolysate mixture and use, but the present invention preferably uses the protein hydrolysate mixture as feed addictive.

The present invention further comprises the steps:

The preparation of protein peptides: the protein enzymatic hydrolyzate that step 3) is obtained under 3000～6000rpm centrifugal 10～30 minutes, the supernatant of telling passes through ultrafiltration membrance filter, cross ultrafiltration membrane system by Fen Liang ≦ 10000Da, the filtrate that obtains is concentrated to 50% of original volume through negative-pressure vacuum, enter step 5) again and carry out spray-drying, make protein peptide powder.

Can be used as feed addictive by the prepared protein peptide powder of the above step of the present invention uses.

The protein hydrolysate mixture and the protein peptide powder that make by preparation method of the present invention have following effect or purposes as feed addictive:

1. the raising efficiency of feed utilization reduces production costs.Active Toplink improves the absorption of animal to nutriment, thereby improves the trans-utilization rate of feed;

2. obviously improve the laying rate of egg fowl, prolong egg-laying peak;

3. improve the weightening finish of poultry, reduce feedstuff-meat ratio, improve meat chicken production performance, improve the albumen apparent digestibility;

4. can reduce incidence, the reduction piglet feedstuff-meat ratio of young animal diarrhoea;

5. be used for aquatic livestock, can improve the weightening finish of fish, reduce feed coefficient, reduce the intestinal contents viscosity, improve the apparent digestibility of total apparent digestibility, protein and the fat of aquatic livestock.

The product that this method makes as the using method of feed addictive is:

(1) using method of protein hydrolysate mixture: the protein hydrolysate mixture is mixed or spice with feed, wherein

Piglet, meat chick, cowboy, lamb, consumption 200～300 gram/ton feeds;

Sow, growing and fattening pigs, milk cow, fattening sheep and ox, consumption 300～1000 gram/ton feeds;

Laying hen, duck, goose, consumption 250～800 gram/ton feeds;

Aquatic livestock, consumption 200～800 gram/ton feeds.

(2) using method of protein peptide powder: protein peptide powder is mixed or spice with feed, wherein

Piglet, meat chick, cowboy, lamb, consumption 150～200 gram/ton feeds;

Sow, growing and fattening pigs, milk cow, fattening sheep and ox, consumption 200～800 gram/ton feeds;

Laying hen, duck, goose, consumption 150～600 gram/ton feeds;

Aquatic livestock, consumption 100～600 gram/ton feeds.

Compared with the prior art the present invention has following superiority:

(1) non-starch polysaccharide enzyme and protease is composite, can overcome prior art and ferment consuming time longly, and the drawback that hydrolysis is single, degree of hydrolysis is low can improve proteolysis speed and hydrolysis degree, improves efficiency of feed utilization, reduces production costs;

(2) interpolation of non-starch polysaccharide enzyme can reduce the content of SNSP in the feed, improves the content of reduced sugar in the hydrolysate, improves the digestibility of animal to feed, improves animal to energy utilization ratio;

(3) technology is simple, and preparation process time is short, is suitable for industrial applications.

, it should be understood that described embodiment only is for the present invention is described, rather than limit the scope of the invention by any way more specific description the present invention by the following example.

The specific embodiment

Embodiment 1 protein hydrolysate mixture-H1

1) pretreatment of raw material:

Enzymolysis substrate-soybean protein isolate is crushed to 100 orders through pulverizer, put into enzymatic vessel, add entry, add the weight of entry and the weight portion proportioning of the contained protein of enzymolysis substrate is 100: 5, stir, transfer pH to 7, in enzymatic vessel, soaked 1 hour under 30 ℃ of conditions, then, carry out 20 minutes sterilization treatment under 80 ℃, sterilized solution cools to 50 ℃, prepares to add enzyme hydrolysis;

2) enzymolysis processing of complex enzyme:

In the pretreated raw material that step 1) obtains, add non-starch polysaccharide enzyme (NSP enzyme) successively, protease is hydrolyzed, temperature is 45 ℃ between the stage of reaction, the pH nature, enzymolysis 14h stopped reaction at 20% o'clock to degree of hydrolysis (DH%);

The addition of various enzymes is: pectase 0U/g albumen, cellulase 0U/g albumen, bacillus licheniformis 2709 alkali protease 5000U/g albumen;

3) enzyme that goes out:

With step 2) liquid after the enzymolysis processing that obtains is in 90 ℃ of enzymes 30 minutes of going out, and flow out liquid and be protein enzymatic hydrolyzate;

4) concentrate:

Protein enzymatic hydrolyzate with step 3) obtains is concentrated into solid content 15～35% through economic benefits and social benefits falling liquid film vacuum concentrator, gets concentrate;

5) spray-drying: concentrate is squeezed into the press spray drying tower through high-pressure pump, 170～190 ℃ of air intakes, 70～85 ℃ of air drafts, temperature is 100～110 ℃ in the tower, carries out spray-drying, finally obtains protein hydrolysate mixture-H1.

Degree of hydrolysis (Degree of Hydrolysis, assay method DH%) are conventional ninhydrin, now simply are described below:

1. the drafting of calibration curve: at first prepare the standard glycine solution, its concentration is 20ug/ml; Then according to the listed application of sample amount order of table 1 application of sample.

The application of sample amount of table 1 glycine, ninhydrin and ethanol

Press the listed application of sample amount of table 1, in tool plug test tube, add standard glycine solution, distilled water and ninhydrin developer successively, in boiling water, heat 15min, in cold water, cool off 5min then; Add 40% ethanolic solution cessation reaction again, place 15min; Return to zero in the mensuration A (absorbance) of 570nm place value with the blank pipe.

2. in the protein hydrolysate mixture-the NH2 Determination on content

Get a certain amount of protein hydrolysate mixture centrifugal 20min under 4000rpm, get supernatant and suitably dilute, make the OD value between 0～1.Absorption 2ml dilution is in tool plug test tube and add 1ml ninhydrin developer, heats 15min behind the mixing in boiling water; In cold water, cool off 5min then; The ethanolic solution cessation reaction that adds 5ml40% again, behind the placement 15min, to be measured.At last, utilize calibration curve to calculate in the protein hydrolysate mixture-content (umol/ml) of NH2.

3. degree of hydrolysis (DH%) calculates

In following formula:

The relative glycine concentration of protein hydrolysate mixture (being protein hydrolyzate in the formula): after being meant the absorbance of the protein hydrolysate mixture of measuring as stated above, the glycine concentration that converses by calibration curve;

The protein concentration of enzymolysis substrate: the protein concentration that is meant enzymolysis substrate (as soybean protein etc.);

75.07: the molecular weight that is glycine;

7.8mmol/g: be the peptide bond number (constant) after the soybean protein complete hydrolysis;

Ho (mmol/g): be the peptide bond number (constant) in the fermentation substrate (raw material).

Embodiment 2 protein hydrolysate mixture-H2

1) pretreatment of raw material:

Enzymolysis substrate-dregs of beans is crushed to 100 orders through pulverizer, put into enzymatic vessel, add entry, add the weight of entry and the weight portion proportioning of the contained protein of enzymolysis substrate is 100:16, stir, transfer pH to 7, in enzymatic vessel, soaked 0.5 hour under 37 ℃ of conditions, then, carry out 10 minutes sterilization treatment under 100 ℃, sterilized solution cools to 60 ℃, prepares to add enzyme hydrolysis;

2) enzymolysis processing of complex enzyme:

In the pretreated raw material that step 1) obtains, add non-starch polysaccharide enzyme (NSP enzyme) successively, protease is hydrolyzed, temperature is 45 ℃ between the stage of reaction, the pH nature, enzymolysis 24h stopped reaction at 42% o'clock to degree of hydrolysis (DH%);

The addition of various enzymes is: pectase 5000U/g albumen, cellulase 2000U/g albumen, bacillus licheniformis 2709 alkali protease 5000U/g albumen, hay bacillus 1398 neutral proteinase 5000U/g albumen;

3) enzyme that goes out:

With step 2) liquid after the enzymolysis processing that obtains is in 120 ℃ of enzymes 15 minutes of going out, and flow out liquid and be protein enzymatic hydrolyzate;

4) concentrate:

Enzymolysis liquid with step 3) obtains is concentrated into solid content 15～35% through economic benefits and social benefits falling liquid film vacuum concentrator, gets concentrate;

5) spray-drying:

Squeeze into the press spray drying tower with concentrating good concentrate through high-pressure pump, 170～190 ℃ of air intakes, 70～85 ℃ of air drafts, temperature is 100～110 ℃ in the tower, carries out spray-drying, finally obtains protein hydrolysate mixture-H2.

Embodiment 3 protein hydrolysate mixture H2 reduced sugar burst sizes

(1) reducing sugar test

1. the drafting of calibration curve

Draw acetate-sodium acetate buffer (pH5.5) 40MI, add DNS reagent 5.0ml, boiling water heating 5min is cooled to room temperature with running water, and the water constant volume is made the standard blank solution to 25ml.

Draw glucose solution 1.00ml, 2.00ml, 3.00ml, 4.00ml, 5.00ml, 6.00ml, 7.00ml respectively, be settled to 100ml with buffer solution respectively, being mixed with concentration is 0.10mg/ml～0.70mg/ml glucose standard liquid.

Draw each 2.00ml of glucose standard liquid (do two parallel) of above-mentioned concentration series respectively, join respectively in the scale test tube, add 2ml water and 5mlDNS reagent more respectively, electromagnetic viscosimeter 3s, boiling water heating 5min, then.Use the running water cool to room temperature, water is settled to 25ml again, serves as the contrast zeroing with the blank sample of standard, measures absorbance OD value at the 540nm place.

With the concentration of glucose is Y-axis, and absorbance OD value is an X-axis, the drawing standard curve.The DNS reagent of each new preparation all needs to repaint calibration curve.

2. measure

Will be through the dregs of beans protein hydrolysate mixture of NSP enzyme enzymolysis in 4000r/min from 20min, through suitably making reduced sugar liquid to be measured after the dilution;

3. result and calculating

(2) add the influence of SNSP to the reduced sugar burst size

Method is with embodiment 2, and result of the test is as shown in table 2.

Table 2, NSP enzyme additive effect

Group	Pectase (U/g)	Lignose enzyme (U/g)	Alkali protease (U/g)	Neutral proteinase (U/g)	Reduced sugar amount (mg/ml)	DH/％
							Blank group	0	0	5000	5000	274.9	30
Test group	5000	2000	5000	5000	558.1	42

Containing a large amount of SNSP (NSP) in the dregs of beans is the principal element that the restriction dregs of beans uses in feed.Can eliminate or weaken the illeffects of these ANFs from the method for external enzymolysis.From this result of the test, behind the use SNSP, the amount of reduced sugar has tangible rising, and the declaratives SNSP is degraded.Simultaneously, add SNSP and can also promote proteoclastic degree and speed.

Embodiment 4 protein hydrolysate mixture-H3

1) pretreatment of raw material:

Enzymolysis substrate-rapeseed dregs is crushed to 80 orders through pulverizer, put into enzymatic vessel, add entry, add the weight of entry and the weight portion proportioning of the contained protein of enzymolysis substrate is 100:10, stir, transfer pH to 8, in enzymatic vessel, soaked 1 hour under 30 ℃ of conditions, then, carry out 15 minutes sterilization treatment under 195 ℃, sterilized solution cools to 55 ℃, prepares to add enzyme hydrolysis;

2) enzymolysis processing of complex enzyme:

In the pretreated raw material that step 1) obtains, add non-starch polysaccharide enzyme (NSP enzyme) successively, protease is hydrolyzed, temperature is 55 ℃ between the stage of reaction, the pH nature, enzymolysis 14h stopped reaction at 28% o'clock to degree of hydrolysis (DH%);

The addition of various enzymes is: cellulase 2000U/g albumen, zytase 4000U/g albumen, bacillus licheniformis 2709 alkali protease 8000U/g albumen, hay bacillus 1398 neutral proteinase 2000U/g albumen;

3) enzyme that goes out:

With step 2) liquid after the enzymolysis processing that obtains is in 120 ℃ of enzymes 15 minutes of going out, and flow out liquid and be protein enzymatic hydrolyzate;

4) concentrate:

Enzymolysis liquid with step 3) obtains is concentrated into solid content 15～35% through economic benefits and social benefits falling liquid film vacuum concentrator, gets concentrate;

5) spray-drying:

Squeeze into the press spray drying tower with concentrating good concentrate through high-pressure pump, 170～190 ℃ of air intakes, 70～85 ℃ of air drafts, temperature is 100～110 ℃ in the tower, carries out spray-drying, finally obtains protein hydrolysate mixture-H3.

Embodiment 5～12, protein hydrolysate mixture-H4～H11

Table 3, protein hydrolysate mixture

The protein hydrolysate mixture	The preparation method	The enzymolysis substrate	Used enzyme or bacterial classification class	Degree of hydrolysis
					H5	With embodiment 2	Dregs of beans	Pectase, cellulase, hydrolysising protease (Alcalase)	22
H6	With embodiment 4	Rapeseed dregs	Zytase, cellulase, compound protease (Protamex)	24
					H7	With embodiment 2	Soya-bean cake	Pectase, cellulase, flavor protease (Flavourzyme)	34
H8	With embodiment 4	The cottonseed dregs of rice	Zytase, cellulase, actinomyces 166	30
					H9	With embodiment 1	Soybean protein isolate	Aspergillus terricola 3942	35
H10	With embodiment 1	FSPC	Aspergillus niger 3350	34
					H11	With embodiment 4	Peanut dregs	Zytase, cellulase, bacillus licheniformis 2709 alkali proteases	28
H12	With embodiment 4	Sunflower seed dregs	Zytase, cellulase, bacillus licheniformis 2709 alkali proteases	27

Embodiment 13 protein peptide powders-T13

The protein enzymatic hydrolyzate through the enzyme that goes out that embodiment 1 obtains was removed slag in centrifugal 10～30 minutes through 3000～6000rpm again, the supernatant of telling passes through ultrafiltration membrance filter, cross ultrafiltration membrane system by Fen Liang ≦ 10000Da, the filtrate that obtains is concentrated to 50% of original volume through negative-pressure vacuum, liquid is got back to enzymolysis process continuation enzymolysis on the film, filtrate then enters step 5) and carries out spray-drying, makes protein peptide powder-T13.

Embodiment 14 protein peptide powders-T14

The protein enzymatic hydrolyzate through the enzyme that goes out that embodiment 2 obtains was removed slag in centrifugal 10～30 minutes through 3000～6000rpm again, the supernatant of telling passes through ultrafiltration membrance filter, cross ultrafiltration membrane system by Fen Liang ≦ 10000Da, the filtrate that obtains is concentrated to 50% of original volume through negative-pressure vacuum, liquid is got back to enzymolysis process continuation enzymolysis on the film, filtrate then enters step 5) and carries out spray-drying, makes protein peptide powder-T14.

Embodiment 15 protein peptide powders-T15

The protein enzymatic hydrolyzate through the enzyme that goes out that embodiment 4 obtains was removed slag in centrifugal 10～30 minutes through 3000～6000rpm again, the supernatant of telling passes through ultrafiltration membrance filter, cross ultrafiltration membrane system by Fen Liang ≦ 10000Da, the filtrate that obtains is concentrated to 50% of original volume through negative-pressure vacuum, and liquid is got back to enzymolysis process continuation enzymolysis on the film.Filtrate then enters step 5) and carries out spray-drying, makes protein peptide powder-T15.

Embodiment 16～23 protein peptide powders-T16～T23

Table 4, protein peptide powder

Protein peptide powder	The preparation method	The enzymolysis substrate	Used enzyme or bacterial classification class
				T-16	With embodiment 5	Dregs of beans	Pectase, cellulase, hydrolysising protease (Alcalase)
T-17	With embodiment 6	Rapeseed dregs	Zytase, cellulase, compound protease (Protamex)
				T-18	With embodiment 7	Soya-bean cake	Pectase, cellulase, flavor protease (Flavourzyme)
T-19	With embodiment 8	The cottonseed dregs of rice	Zytase, cellulase, actinomyces 166
				T-20	With embodiment 9	Soybean protein isolate	Aspergillus terricola 3942
T-21	With embodiment 10	FSPC	Aspergillus niger 3350
				T-22	With embodiment 11	Peanut dregs	Zytase, cellulase, bacillus licheniformis 2709 alkali proteases
T-23	With embodiment 12	Sunflower seed dregs	Zytase, cellulase, bacillus licheniformis 2709 alkali proteases

The protein hydrolysate mixture that embodiment 24 use the inventive method make is as the application of feed addictive in Swine Production

Be the effect of check product of the present invention and the effect on pig industry, and replace antibiotic feasibility, test is chosen body weight and is divided into 3 processing near 36 healthy of 24 age in days weanling pigs (male and female half and half), each processing (group) 12 repeats, each repeats 6 pigs, be 3 public 3 mothers, 21 days experimental periods,, result of the test is listed in table 5.

Three processing are as follows:

Blank group: do not add any additives, the pig basal diet of only feeding;

The antibiotic control group: pulvis aureomycin 100mg/kg evenly is mixed in the basal diet;

The protein peptides group: use protein hydrolysate mixture-H2 of embodiment 2, consumption 300mg/kg evenly is mixed in basal diet as feed addictive.

Table 5, the result of use of protein hydrolysate mixture in Swine Production

Processing/repetition	The blank group	Antibiotic control group (100mg/kg)	Protein peptides group (300mg/kg)	P
					Full phase F/G	1.850±0.214	1.710±0.340	1.617±0.098	<0.05
Full phase diarrhoea index	0.327±0.224	0.069±0.205	0.107±0.134	<0.05

As can be seen from Table 3, added the protein peptides group of protein hydrolysate mixture, feedstuff-meat ratio reduces, and the incidence of grice diarrhoea reduces.

Embodiment 25 the inventive method make protein peptide powder as the influence of feed addictive to meat chick production performance (speed of weight increment and feed consumption)

Ai Wei mattress meat chick is divided into 3 groups, and each component becomes 4 repetitions, and each repeats 20 chickens, 7 weeks of examination phase.Processed group 3 is fed protein peptide powder-T14 of embodiment 14 as feed addictive, and experimental design sees Table 6.

Table 6, feeding of broiler experimental design

Group	Disposition	Active ingredient (g/ ton feed)
			A	Blank	0
B	Antibiotic contrast (aureomycin)	150
			C	Protein peptide powder	600

When 1 Japanese instar chickling goes into to give up and 4 age ends (28 age in days) in week, 7 age ends in week (49 age in days) weigh respectively, and statistics feed consumption rate calculates dead number of elements, weightening finish, feed intake and the feedstuff-meat ratio in 0～4 week and 4～7 all ages.The results are shown in Table 7.

Table 7,49 ages in days add the influence of protein peptide powder to meat chick production performance

Group	Weightening finish (kg)	Feed intake (kg)	Feedstuff-meat ratio
				A	2.307±0.087b	5.211±0.014	2.230±0.068b
B	2.350±0.097a	5.175±0.005	2.219±0.090a
				C	2.375±0.061a	5.152±0.011	2.178±0.006a

Annotate: the right shoulder of same column indicates different letter representation significant differences

The data of table 7 show, add protein peptide powder as the weightening finish that feed addictive can obviously improve fryer, also can improve feed efficiency, thereby save the feed spending, increase economic efficiency.

Protein hydrolysate mixture-H2 that embodiment 26 use the inventive method make is as the application of feed addictive in layer breeding

For check protein hydrolysate mixture as feed addictive in result of use on the laying hen and the application prospect in animal husbandry, Luo Man laying hen later stage chicken 2000 plumages are selected in test for use, divide two groups, every group of 1000 laying hens, feed with 2 kinds of different daily rations, promptly the A group is the blank group, and B organizes feed per ton and adds protein hydrolysate mixture-H2 of 800 embodiment 2 that restrain as feed addictive, 2 months experimental periods, the results are shown in table 8.The detection index is rate of broken eggs (%), laying rate (%), egg size (a gram/egg), death rate (%), feed efficiency (feed consumption g/kg egg).

Table 8, the application of protein hydrolysate mixture in layer breeding

	The A group	The B group	p
				Later stage laying rate of chicken (%)	80.11±2.84	84.54±2.51	<0.05

Feedstuff-egg ratio (F/E)	2.68±0.25	2.61±0.42	<0.05
				Rate of broken eggs (%)	3.07±0.43	2.21±0.24	<0.05
Death rate %	1.51±0.26	0.80±0.28	<0.05
				Egg size (g)	64.71±0.58	66.75±0.51	<0.05

As can be seen from Table 7, adding the protein hydrolysate mixture in egg feedstuff can obviously improve egg fowl laying rate, increases egg size, reduce feedstuff-egg ratio, rate of broken eggs as feed addictive, reduce death rate, thereby improve efficiency of feed utilization, production performance and reduce production costs.

The effect that embodiment 27 protein peptide powders-T14 feeds aquatic livestock

Test fish average weight 100g, test is carried out in indoor aquarium.The duration of test water quality parameter: 25 ± 10 ℃ of water temperatures, dissolved oxygen 5～7mg/L, pH6.8～7.5, ammoniacal nitrogen are less than 0.2mg/L.Test 2 processing, be divided into control group and test group, each handles three aquariums (three repetitions), each aquarium 10 tail fish, experimental design and result such as table 8.

The trial test of test fish elder generation is assigned in 2 groups after 20 days at random, changes the formal test phase over to.Experimental period, the basal diet prescription is Gao Song (1980), Takeuchi recommended levels such as (1980), test group is fed protein peptide powder-T14 of embodiment 14 as feed addictive on the basal diet basis, addition is a 500g/t material, bait throwing in every day 3 times, the morning 8:00, noon 1:00, afternoon 6:00, daily ration, feeding quantity be fish heavy 3～5%, 50 days experimental periods.

Table 9, Tilapia mossambica production performance index and enteron aisle viscosity change

Detect index	Control group	Test group	p
				Average weight gain	56.10±1.48	75.57±2.39	<0.05
Feed coefficient	2.79±0.06	2.13±0.04	<0.05
				Relative viscosity	1.197±0.05	1.027±0.02	<0.05

By this test explanation, protein peptide powder not only can improve the production performance of aquatic livestock as feed addictive, and can reduce the viscosity of its enteron aisle.

Claims (5)

1, a kind of preparation method of novel protein feed additive is characterized in that comprising the steps:

1) pretreatment of raw material:

The enzymolysis substrate is crushed to 40～100 orders, put into enzymatic vessel, add entry, add the weight of entry and the weight proportion of the contained protein of enzymolysis substrate is 100:5～18, stir, transfer pH to 7～9, soaked 0.5～1 hour under 30～37 ℃ of conditions, then, 80～100 ℃ of down sterilizations 10～20 minutes, after cool to 50～60 ℃;

2) enzymolysis processing of complex enzyme:

In the pretreated raw material that step 1) obtains, add non-starch polysaccharide enzyme (NSP enzyme) successively, protease is hydrolyzed, 45～60 ℃ of temperature, the pH nature, enzymolysis 6～14h stopped reaction at 20～42% o'clock to degree of hydrolysis (DH%);

3) enzyme that goes out:

With step 2) liquid after the enzymolysis processing that obtains is in 90～120 ℃ of enzymes 15～30 minutes of going out, and flow out liquid and be protein enzymatic hydrolyzate;

4) concentrate:

With the protein enzymatic hydrolyzate that step 3) obtains, be concentrated into solid content 15～35%, get concentrate;

5) spray-drying:

The concentrate of step 4) is squeezed into the press spray drying tower through high-pressure pump, 170～190 ℃ of air intakes, 70～85 ℃ of air drafts, temperature is 100～110 ℃ in the tower, carries out spray-drying, obtains the protein hydrolysate mixture.

2, the preparation method of novel protein feed additive as claimed in claim 1 is characterized in that described enzymolysis substrate is following vegetable protein: any in soybean protein isolate, FSPC, dregs of beans, soya-bean cake, cotton cake dregs, dregs of rapeseed cake, sunflower seeds grouts, the peanut dregs, two or more mixture.

3, the preparation method of novel protein feed additive as claimed in claim 1, it is characterized in that described non-starch polysaccharide enzyme be in pectase, cellulase, the zytase any, two or more; Described protease is neutral proteinase and/or alkali protease, and protease can be following bacterium: any in protease that hay bacillus 1398, actinomyces 166, aspergillus terricola 3942, aspergillus niger 3350 and/or bacillus licheniformis 2709 produce and hydrolysising protease (Alcalase), flavor protease (Flavourzyme), the compound protease (Protamex), two or more.

4, the preparation method of novel protein feed additive as claimed in claim 3 is characterized in that the addition of described pectase, cellulase, zytase and protease is: pectase, 0～3000U/g albumen; Cellulase, 0～2000U/g albumen; Zytase, 0～4000U/g albumen; Protease, 1000～10000U/g albumen.

5, the preparation method of novel protein feed additive as claimed in claim 1 is characterized in that further comprising the steps:

The preparation of protein peptides: the protein enzymatic hydrolyzate that step 3) is obtained under 3000～6000rpm centrifugal 10～30 minutes, the supernatant of telling passes through ultrafiltration membrance filter, cross ultrafiltration membrane system by Fen Liang ≦ 10000Da, the filtrate that obtains is concentrated to 50% of original volume through negative-pressure vacuum, enter step 5) again and carry out spray-drying, make protein peptide powder.