CN109136315A - A kind of heme oligopeptides microcapsule powder and preparation method thereof - Google Patents
A kind of heme oligopeptides microcapsule powder and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to slaughtering processing byproducts to comprehensively utilize processing technique field, disclose a kind of heme oligopeptides microcapsule powder and preparation method thereof.This method using it is being collected on the broiler chicken slaughter line for meeting food security standard, through anticoagulant broiler chicken new blood as raw material, comprising the following steps: (1) the swelling broken wall of erythrocyte；(2) ferrohemoglobin is protected；(3) ferrohemoglobin complex enzyme hydrolysis；(4) ultra-filtration and separation purifies；(5) prepared by heme oligopeptides microcapsules, obtains the heme oligopeptides microcapsule powder that molecular mass is less than 3k Da, heme yield is high, stability is good.
Description
Technical field
The invention belongs to slaughtering processing byproducts to comprehensively utilize processing technique field, and in particular, to a kind of ferrous iron
Ferroheme oligopeptides microcapsule powder and preparation method thereof.
Background technique
Iron is the essential trace elements of the human body, and asiderosis or iron Use barriers will lead to oxygen transportation and storage capability subtracts
Weak, long-time iron deficiency can exhaust body reserve, reduce hemoglobin level and hematocrit value, and iron-deficiency anemia occurs
(iron deficiency anemia, IDA).IDA is worldwide generally existing, according to world public health nutritive issue
Data show that there are about 24.8% populations to be perplexed by IDA in the world, and therefore, IDA is still the health problem urgently paid close attention at present.It lacks
Iron is body accumulation for a long time as a result, hypoferric anemia is once caused to be difficult to supplement by food and restored completely, mesh
Former world mainly assists in the treatment of iron-deficiency anemia with iron supplementary, to alleviate iron deficiency symptoms, mitigates the damage of body.Just at present
Common iron supplementary, although having certain remission effect to hypoferric anemia symptom, there are still problems once: biological utilisation
Rate is low, gastrointestinal side effect is big, and dietetic contraindication is more, and free iron toxicity is big；Iron taste weight poor taste, treatment cycle length etc..
Livestock and poultry blood is slaughtering processing latter by-product, rich in nutritional ingredients such as protein, iron, but due to collecting
The missing of technology, evaluation and exploration technology etc., on the one hand it is the pole of resource that the livestock and poultry blood of the overwhelming majority, which is still discarded, at present
Big waste, on the other hand since discarding for livestock and poultry blood brings great pressure to environmental protection treatment, livestock and poultry blood is opened
It is that fowl butchers processing enterprise's key technology extremely to be solved that hair, which utilizes,.Blood is rich iron-containing living resources, can be used for the benefit of people
Chalybeate.Have obvious trophic function to human body in livestock and poultry blood is heme, but since the hemoglobin in blood is in people
It is difficult to be digested absorption in body alimentary canal, therefore, livestock and poultry blood is seldom used as iron supplementary at present.To improve and solving animal blood
The utilization rate of liquid, the country have carried out a large amount of research, and current research is concentrated mainly on the processing of hemoglobin enzymatic hydrolysis, to improve blood
The utilization rate of Lactoferrin.
The patent of invention of Publication No. CN101649342B disclose a kind of bivalent heme polypeptide-ferrum purification process and its
Purposes.The core content of the patent disclosure includes: 1. supercritical ultrasonics technology broken wall erythrocyte；2. being digested in a vacuum chamber using pancreatin
Hemoglobin discharges bivalent heme polypeptide-ferrum, and vacuum (0.01MPa) high temperature (95 DEG C) enzyme deactivation；3. by enzymolysis liquid centrifuging and taking
Clear liquid isolates and purifies bivalent heme polypeptide-ferrum using metal chelate affinity chromatography or electromagnetism." pass through vacuum high-frequency described in the patent
Impulse electric field enzymatic hydrolysis, vacuum high-temperature enzyme deactivation " is to obtain the big iron of bivalent heme.On the one hand, which does not describe the Asia of product
On the other hand the protecting effect of iron yield and ferroheme ferrous iron " maintains vacuum degree 0.01MPa, increases true mentioned by the patent
Empty room temperature is to 95 DEG C, high temperature enzyme deactivation 10-10min ", and under this vacuum condition, temperature is can not to reach 95 DEG C completely.
The patent of invention of Publication No. CN102150837 discloses ferrous heme-enriched peptide formula of chewable tablets and production method.
The core of the patent be using neutral proteinase and flavor protease enzymatic hydrolysis hemoglobin, after enzymatic hydrolysis respectively through 10kD and
3kD ultrafiltration membrane ultrafiltration obtains heme peptide solution；Enzymolysis liquid ultrafiltrate is spray-dried to be made Gly-His-Lys.The patent is through enzyme
Does is products obtained therefrom " ferrous heme-enriched peptide " after solution, ultrafiltration, spray drying? because unprotected hemoglobin is in aerobic item
It is digested under part, ultrafiltration and spray drying, heme are easily oxidized as high-speed rail (ferric iron) ferroheme, final institute
The product obtained can not be " ferrous heme-enriched peptide ", and also not divide products therefrom " ferrous heme-enriched peptide " content in patent
It analyses, is only added to " 0.5%~1.5% ascorbic acid " in formula when prepared by " chewable tablets " as antioxidant in patent
Ferrous heme-enriched peptide is protected, but if ferrous heme-enriched peptide has been oxidized to " protoferriheme peptide " during the preparation process, in " nozzle
Chew piece " in ascorbic acid " protoferriheme peptide " can not be reduced to " ferrous heme-enriched peptide ", therefore, in the patent
" ferrous heme-enriched peptide " content in " ferrous heme-enriched peptide chewable tablets " is extremely limited.
The patent of invention of Publication No. CN201611134138.6 discloses a kind of from animal blood extraction small peptide ferrous iron blood
The production method of red pigment, the core content of the patent mainly include following several aspects: 1) being separately added into 2%~15% before taking a blood sample
Anti-coagulants and 0.045%~0.048% ascorbic acid as antioxidant；2) multigelation method is assisted using ultrasonic wave
Carry out blood haemolysis；3) inhibit the oxidation of heme in enzymolysis process using inert gas；4) it digests and is added in solution
Ascorbic acid and casein phosphopeptide carry out anti-oxidant, spray drying acquisition small peptide heme；5) with concentration be 21~
25% (80~100 DEG C) hydrolysis plasma proteins of concentrated hydrochloric acid high temperature, reflux crystallization obtain small peptide heme crystal；It 6) will spray
The dry enzymatic hydrolysis " small peptide heme " of mist is milled after mixing with " the small peptide heme crystal " of concentrated hydrochloric acid hydrolysis, mistake
Sieve, obtains powdered small peptide heme finished product.It is unreasonable using 2%~15% anti-coagulants described in patent；Enzyme
Only with addition ascorbic acid and casein phosphorus in inert gas (specific type is not specified), spray-drying process in solution preocess
Sour peptide cannot effectively inhibit the oxidation of heme；It is that impossible obtain small peptide heme after plasma protein hydrolysis
's.The content of " small peptide heme finished product " the Small Peptides heme not described in patent is how many.
Master's thesis " development and its Study on functional properties of food-grade ferrous heme-enriched peptide " uses different type protective agent
Ox blood enzymatic hydrolysis is made an addition to respectively to produce in the enzymatic hydrolysis substrate and product of ferrous heme-enriched peptide, examines or check it to ferrous protective value.
In enzyme digestion reaction, optimal antioxidant/reducing agent group is combined into ascorbic acid/anhydrous sodium sulfite (2:1), and ferrous content mentions
It is high by 24.5%.In enzymolysis product, optimal antioxidant/reducing agent group is combined into ascorbic acid/anhydrous sodium sulfite (2:1),
Ferrous content improves 37.7%.The ferrous protecting effect that antioxidant is added in enzymolysis product, which is an advantage in enzyme digestion reaction, to be added
Antioxidant.The guard method of ferroheme ferrous iron is by adding antioxidants ascorbic acid in enzyme digestion reaction described in paper
And anhydrous sodium sulfite, these antioxidant stability at later stage are not high, and antioxidant effect is undesirable.
Summary of the invention
Place in order to overcome the shortcomings and deficiencies of the prior art, the primary purpose of the present invention is that providing a kind of effective
The preparation method of heme oligopeptides microcapsule powder.This method on yellow feather dwarf broiler slaughter production line to collect through anticoagulant
Chicken blood be raw material, haemocyte broken wall is carried out with preferred water-swellable method and discharges hemoglobin, prepares hemoglobin before enzymatic hydrolysis
It is obtained for stable histidine-ferrohemoglobin through the technological means such as complex enzyme hydrolysis, ultrafiltration purification separation, microcapsule embedded
The heme oligopeptides microcapsule powder that molecular mass is less than 3k Da, heme yield is high, stability is good.
Another object of the present invention is to provide a kind of micro- glue of heme oligopeptides that above-mentioned preparation method is prepared
Capsule powder.
A kind of preparation method of heme oligopeptides microcapsule powder, comprising the following steps:
S1. the swelling broken wall of erythrocyte: the new blood for the yellow-feather broiler that slaughter production line is butchered is collected, anti-coagulants is added
Anticoagulation is carried out, then anticoagulant new blood is centrifuged, abandoning supernatant blood plasma, the physiological saline cleaning of the quality such as addition haemocyte,
Centrifugation, is repeated 3 times, and obtains blood rbc, and the clean water of 3~4 times of volumes of blood rbc is added, it is sufficiently stirred 20~
30min makes erythrocyte swelling rupture, releases hemoglobin；
S2. the protection of ferrohemoglobin: by broken wall erythrocyte obtained by S1, be added erythrocyte quality 0.04~
0.06% histidine stands 30~60mim of reaction after mixing evenly, obtains histidine-ferrohemoglobin compound；
S3. histidine-ferrohemoglobin compound complex enzyme hydrolysis: histidine-ferrohemoglobin obtained by S2 is multiple
It closes object and first carries out alkali protease enzymatic hydrolysis, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 4~6%, basic protein
Enzyme dosage 3~8kU/g protein, pH value 8.5, temperature 45 C digest 5h with this condition, then with hydrochloric acid by enzymolysis liquid pH value
6.0 are adjusted to, flavor protease is added by 1~3kU/g protein, after digesting 3h, obtains ferrohemoglobin enzymolysis liquid, is cooled to
4 DEG C of inhibitory enzyme activities；
S4. ultra-filtration and separation purifies heme oligopeptides: using 4000rpm by ferrohemoglobin enzymolysis liquid obtained by S3 from
The heart removes wherein large particulate matter, then by the ferrohemoglobin enzymolysis liquid after centrifugation under 0.5MPa pressure, with surpassing for 3kD
Filter membrane carries out hyperfiltration treatment and removes enzyme and macromolecular substances, obtains the heme oligopeptides concentrate that molecular mass is less than 3kD；
S5. prepared by heme oligopeptides microcapsule powder: being added into heme oligopeptides concentrate obtained by S4 porous
For starch as core material, the dosage of porous-starch is 1.5 times of dry biomass in heme oligopeptides concentrate；In 40 DEG C of items
It is sufficiently adsorbed to saturation under part, adds 1.6 times of core material quality of Arabic gum as wall material, ultrasonic wave auxiliary mixing embedding,
Heme oligopeptides microcapsule powder is made through vacuum freeze drying.
The dosage of clean water described in step S1 is 3 times of volumes of blood rbc, mixing time 30min.
The additional amount of histidine described in step S2 is the 0.05% of erythrocyte quality.
The condition of enzymatic hydrolysis described in step S3 is the protein content of substrate mass concentration 5%, basic protein enzyme dosage 5kU/g
Protein；The additive amount of the flavor protease is 2kU/g.
Ferrohemoglobin degree of hydrolysis is up to 27.3~34.2% in ferrohemoglobin enzymolysis liquid obtained by step S3, ferrous blood
Red pigment peptide yield is 82.6~87.5.
The content of heme oligopeptides is 20.8~22.5mg/ in heme oligopeptides concentrate obtained by step S4
ML, the rate of recovery of heme are 83.5~84.1%.
Heme oligopeptides microcapsule powder obtained by step S5, microcapsule embedded rate are 95.83~97.34%, ferrous blood
Red pigment oligopeptides 203~207mg/g of content, heme yield are 77.64~79.33%.
A kind of heme oligopeptides microcapsule powder being prepared by above-mentioned preparation method.Microcapsule powder is in uniform
Grey-brown powder shape, no smell of blood, particle is uniform, non-block phenomenon.
Heme oligopeptides microcapsule powder of the present invention is soluble in water, and ferrous iron of the gained containing mass fraction 0.5% is blood red
After 60 DEG C, 70 DEG C of heating 10min, the yield of the heme oligopeptides in solution is the aqueous solution of plain oligopeptides microcapsule powder
96.3%, the heme rate of recovery is 88.3%, therefore 0.5% heme oligopeptides microcapsule powder solution is being lower than temperature
Under the conditions of 70 DEG C, heme oligopeptides has good stability.
Compared with prior art, the invention has the following beneficial effects:
The present invention passes through convenient, fast, effective water-swellable method broken wall erythrocyte；With histidine to ferrohemoglobin
It is protected, prevents the oxidation of ferrohemoglobin；Ferrohemoglobin enzymolysis product is efficiently prepared in complex enzyme hydrolysis method；
The heme oligopeptide solution for obtaining high concentration is separated with ultra-filtration and separation purification process；It is obtained by the preparation of microcapsule embedded method
Obtain heme oligopeptides content height, heme stabilization, the uniform heme oligopeptides of the particle without smell of blood iron rust taste
Microcapsule powder can be used as the raw material of effective iron-deficiency anemia iron supplementary product.
Specific embodiment
Further the present invention is illustrated below by specific embodiment, but should not be understood as to the scope of the present invention
Limitation, those skilled in the art makes some nonessential changes and adjustment according to the content of foregoing invention, belongs to this
The protection scope of invention.
Raw material used in the following embodiment be collected on the yellow-feather broiler slaughter line for meet food security standard, through anti-
Solidifying broiler chicken new blood, selected alkali protease actual measurement enzyme activity is 172kU/g, flavor protease actual measurement enzyme activity is
147kU/g.Unless stated otherwise, the present invention uses reagent, method and apparatus for the art conventional reagent, method and are set
It is standby.
Embodiment 1
S1. the swelling broken wall of erythrocyte: being centrifuged 10min to anticoagulant fresh chicken blood with 4000rpm, abandon supernatant blood plasma, then
With physiological saline cleaning, the centrifugation of the quality such as haemocyte, it is repeated 3 times, obtains blood rbc, 3 times of bodies of blood rbc are added
Long-pending clean water, 30min, which is sufficiently stirred, makes erythrocyte swelling rupture, obtains the hemoglobin of broken wall, the swelling of erythrocyte
Sporoderm-broken rate reaches 96.6%.
S2. by broken wall hemoglobin described in S1, erythrocyte quality 0.05% protection of ferrohemoglobin: is added
Histidine stands reaction 60mim after mixing evenly, obtains histidine-ferrohemoglobin compound, and histidine-ferrous iron is blood red
The yield of albumen composition is up to 86%.
S3. histidine-ferrohemoglobin compound complex enzyme hydrolysis: histidine-ferrohemoglobin obtained by S2 is multiple
It closes object and first carries out alkali protease enzymatic hydrolysis, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 5%, alkali protease
Dosage 5kU/g protein, pH value 8.5, temperature 45 C, after digesting 5h, tune pH value to 6.0, flavor protease additive amount 2kU/g
After digesting 3h, ferrohemoglobin enzymolysis liquid is obtained, 4 DEG C of inhibitory enzyme activities are cooled to；Ferrohemoglobin degree of hydrolysis reaches
30.1%, heme peptide yield is 85.8%.
S4. ultra-filtration and separation purifying prepares heme oligopeptides concentrate: using 4000rpm by ferrous blood red egg obtained by S3
White enzymolysis liquid centrifugation removal wherein large particulate matter, in order to avoid blocking ultrafiltration membrane, then by the ferrohemoglobin enzymolysis liquid after centrifugation
Under 0.5MPa pressure, hyperfiltration treatment is carried out with the ultrafiltration membrane of 3kD and removes enzyme and macromolecular substances, molecular mass is obtained and is less than
3kD heme oligopeptides concentrate；The content of heme oligopeptides is in gained heme oligopeptides concentrate
21.8mg/mL, the rate of recovery of heme are 83.9%.
S5. prepared by heme oligopeptides microcapsule powder: 1.5 times being added into heme oligopeptides concentrate obtained by S4
The porous-starch of its dry matter content is sufficiently adsorbed to saturation under the conditions of 40 DEG C as core material, adds 1.6 times of core material amount
Heme oligopeptides microcapsule powder is made through vacuum freeze drying as wall material, ultrasonic wave auxiliary mixing embedding in Arabic gum.
Gained heme oligopeptides microcapsule powder, embedding rate 96.54%, heme widow are prepared with embodiment 1
Peptide content 203mg/g, heme yield be 78.45%, microcapsule powder be in uniform grey-brown powder shape, no smell of blood,
Particle is uniform, non-block phenomenon.The aqueous solution of the 0.5% heme oligopeptides microcapsule powder containing mass fraction is heated through 60 DEG C
10min, heme oligopeptides yield is 96.3% in solution, and heme oligopeptides has good stability.
Embodiment 2
S1. the swelling broken wall of erythrocyte: being centrifuged 10min to anticoagulant fresh chicken blood with 4000rpm, abandon supernatant blood plasma, then
With physiological saline cleaning, the centrifugation of the quality such as haemocyte, it is repeated 3 times, obtains blood rbc, 3 times of bodies of blood rbc are added
Long-pending clean water, 30min, which is sufficiently stirred, makes erythrocyte swelling rupture, obtains the hemoglobin of broken wall, the swelling of erythrocyte
Sporoderm-broken rate reaches 95.6%.
S2. by broken wall hemoglobin obtained by S1, erythrocyte quality 0.04% protection of ferrohemoglobin: is added
Histidine stands reaction 60mim after mixing evenly, obtains histidine-ferrohemoglobin compound, and histidine-ferrous iron is blood red
The yield of albumen composition is up to 85%.
S3. histidine-ferrohemoglobin compound complex enzyme hydrolysis: histidine-ferrohemoglobin obtained by S2 is multiple
It closes object and first carries out alkali protease enzymatic hydrolysis, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 4%, alkali protease
Dosage 4kU/g protein, pH value 8.5, temperature 45 C, after digesting 5h, tune pH value to 6.0, flavor protease additive amount 1kU/g
After digesting 3h, ferrohemoglobin enzymolysis liquid is obtained, 4 DEG C of inhibitory enzyme activities are cooled to；Ferrohemoglobin degree of hydrolysis reaches
27.3%, heme peptide yield is 82.6%.
S4. ultra-filtration and separation purifying prepares heme oligopeptides concentrate: using 4000rpm by ferrous blood red egg obtained by S3
White enzymolysis liquid centrifugation removal wherein large particulate matter, in order to avoid blocking ultrafiltration membrane, then by the ferrohemoglobin enzymolysis liquid after centrifugation
Under 0.5MPa pressure, hyperfiltration treatment is carried out with the ultrafiltration membrane of 3kD and removes enzyme and macromolecular substances, molecular mass is obtained and is less than
3kD heme oligopeptides concentrate；The content of heme oligopeptides is in gained heme oligopeptides concentrate
20.8mg/mL, the rate of recovery of heme are 84.1%.
S5. prepared by heme oligopeptides microcapsule powder: 1.5 times being added into heme oligopeptides concentrate obtained by S4
The porous-starch of its dry matter content is sufficiently adsorbed to saturation under the conditions of 40 DEG C as core material, adds 1.6 times of core material amount
Heme oligopeptides microcapsule powder is made through vacuum freeze drying as wall material, ultrasonic wave auxiliary mixing embedding in Arabic gum.
Gained heme oligopeptides microcapsule powder, embedding rate 95.83%, heme widow are prepared with embodiment 2
Peptide content 206mg/g, heme yield be 77.64%, microcapsule powder be in uniform grey-brown powder shape, no smell of blood,
Particle is uniform, non-block phenomenon.The aqueous solution of the 0.5% heme oligopeptides microcapsule powder containing mass fraction is heated through 60 DEG C
10min, heme oligopeptides yield is 97.2% in solution, and heme oligopeptides has good stability.
Embodiment 3
S1. the swelling broken wall of erythrocyte: being centrifuged 10min to anticoagulant fresh chicken blood with 4000rpm, abandon supernatant blood plasma, then
With physiological saline cleaning, the centrifugation of the quality such as haemocyte, it is repeated 3 times, obtains blood rbc, 3 times of bodies of blood rbc are added
Long-pending clean water, 30min, which is sufficiently stirred, makes erythrocyte swelling rupture, obtains the hemoglobin of broken wall, the swelling of erythrocyte
Sporoderm-broken rate reaches 97.2%.
S2. by broken wall hemoglobin obtained by S1, the group of erythrocyte amount 0.06% protection of ferrohemoglobin: is added
Propylhomoserin stands reaction 60mim after mixing evenly, obtains histidine-ferrohemoglobin compound, the blood red egg of histidine-ferrous iron
The yield of white compound is up to 87%.
S3. histidine-ferrohemoglobin compound complex enzyme hydrolysis: histidine-ferrohemoglobin obtained by S2 is multiple
It closes object and first carries out alkali protease enzymatic hydrolysis, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 6%, alkali protease
Dosage 8kU/g protein, pH value 8.5, temperature 45 C, after digesting 5h, tune pH value to 6.0, flavor protease additive amount 3kU/g
After digesting 3h, ferrohemoglobin enzymolysis liquid is obtained, 4 DEG C of inhibitory enzyme activities are cooled to；Ferrohemoglobin degree of hydrolysis reaches
34.2%, heme peptide yield is 87.5%.
S4. ultra-filtration and separation purifying prepares heme oligopeptides concentrate: using 4000rpm by ferrous blood red egg obtained by S3
White enzymolysis liquid centrifugation removal wherein large particulate matter, in order to avoid blocking ultrafiltration membrane, then by the ferrohemoglobin enzymolysis liquid after centrifugation
Under 0.5MPa pressure, hyperfiltration treatment is carried out with the ultrafiltration membrane of 3kD and removes enzyme and macromolecular substances, molecular mass is obtained and is less than
3kD heme oligopeptides concentrate；The content of heme oligopeptides is in gained heme oligopeptides concentrate
22.5mg/mL, the rate of recovery of heme are 83.5%.
S5. prepared by heme oligopeptides microcapsule powder: 1.5 times being added into heme oligopeptides concentrate obtained by S4
The porous-starch of its dry matter content is sufficiently adsorbed to saturation under the conditions of 40 DEG C as core material, adds 1.6 times of core material amount
Heme oligopeptides microcapsule powder is made through vacuum freeze drying as wall material, ultrasonic wave auxiliary mixing embedding in Arabic gum.
Gained heme oligopeptides microcapsule powder, embedding rate 97.34%, heme widow are prepared with embodiment 3
Peptide content 207mg/g, heme yield be 79.33%, microcapsule powder be in uniform grey-brown powder shape, no smell of blood,
Particle is uniform, non-block phenomenon.The aqueous solution of the 0.5% heme oligopeptides microcapsule powder containing mass fraction is heated through 60 DEG C
10min, heme oligopeptides yield is 97.1% in solution, and heme oligopeptides has good stability.
Comparative example
S1. the swelling broken wall of erythrocyte: being centrifuged 10min to anticoagulant fresh chicken blood with 4000rpm, abandon supernatant blood plasma, then
With physiological saline cleaning, the centrifugation of the quality such as haemocyte, it is repeated 3 times, obtains blood rbc, 3 times of bodies of blood rbc are added
Long-pending clean water, 30min, which is sufficiently stirred, makes erythrocyte swelling rupture, obtains the hemoglobin of broken wall.
S2. by broken wall hemoglobin obtained by S1, the Asia of erythrocyte amount 0.01% protection of ferrohemoglobin: is added
Sodium nitrate stands reaction 60mim after mixing evenly, obtains nitroso-ferrohemoglobin compound.
S3. nitroso-ferrohemoglobin compound complex enzyme hydrolysis: by obtained by S2 through nitroso-ferrohemoglobin
Compound first carries out trypsin digestion, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 5%, trypsase are used
5kU/g protein is measured, pH value 7.0, temperature 45 C, after digesting 5h, ferrohemoglobin degree of hydrolysis is up to 18.5%, heme
Peptide yield is 40.2%.
The comparative example protects ferrohemoglobin with sodium nitrite substitution histidine；Alkalinity is substituted with trypsase
Protease and flavor protease since heme peptide yield is lower, therefore do not carry out ultra-filtration and separation purifying and microcapsules preparation.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (8)
1. a kind of preparation method of heme oligopeptides microcapsule powder, it is characterised in that the following steps are included:
S1. the swelling broken wall of erythrocyte: the new blood for the yellow-feather broiler that slaughter production line is butchered is collected, anti-coagulants is added to carry out
Anticoagulation, then anticoagulant new blood is centrifuged, supernatant blood plasma is abandoned, physiological saline cleaning, the centrifugation of the quality such as haemocyte is added,
It is repeated 3 times, obtains blood rbc, the clean water of 3~4 times of volumes of blood rbc is added, 20~30min, which is sufficiently stirred, makes blood
Red blood cell swelling rupture, releases hemoglobin；
S2. by broken wall erythrocyte obtained by S1, erythrocyte quality 0.04~0.06% protection of ferrohemoglobin: is added
Histidine stands 30~60mim of reaction after mixing evenly, obtains histidine-ferrohemoglobin compound；
S3. histidine-ferrohemoglobin compound complex enzyme hydrolysis: by histidine-ferrohemoglobin compound obtained by S2
First carry out alkali protease enzymatic hydrolysis, the condition of enzymatic hydrolysis are as follows: the protein content of substrate mass concentration 4~6%, alkali protease are used
3~8kU/g protein, pH value 8.5 are measured, and temperature 45 C digests 5h with this condition, then enzymolysis liquid pH value is adjusted to hydrochloric acid
6.0, flavor protease is added by 1~3kU/g protein, after digesting 3h, obtains ferrohemoglobin enzymolysis liquid, is cooled to 4 DEG C
Inhibitory enzyme activity；
S4. ultra-filtration and separation purifies heme oligopeptides: being gone the centrifugation of ferrohemoglobin enzymolysis liquid obtained by S3 using 4000rpm
Except wherein large particulate matter, then by the ferrohemoglobin enzymolysis liquid after centrifugation under 0.5MPa pressure, with the ultrafiltration membrane of 3kD into
Row hyperfiltration treatment removes enzyme and macromolecular substances, obtains the heme oligopeptides concentrate that molecular mass is less than 3kD；
S5. prepared by heme oligopeptides microcapsule powder: porous-starch is added into heme oligopeptides concentrate obtained by S4
As core material, the dosage of porous-starch is 1.5 times of dry biomass in heme oligopeptides concentrate；Under the conditions of 40 DEG C
It is sufficiently adsorbed to saturation, adds 1.6 times of core material quality of Arabic gum as wall material, ultrasonic wave auxiliary mixing embedding, through true
Heme oligopeptides microcapsule powder is made in vacuum freecing-dry.
2. preparation method according to claim 1, it is characterised in that: the dosage of clean water described in step S1 is that blood is red thin
3 times of volumes of born of the same parents, mixing time 30min.
3. preparation method according to claim 1, it is characterised in that: the additional amount of histidine described in step S2 is blood red thin
The 0.05% of cytoplasm amount.
4. preparation method according to claim 1, it is characterised in that: the condition of enzymatic hydrolysis described in step S3 is that substrate quality is dense
The protein content of degree 5%, basic protein enzyme dosage 5kU/g protein；The additive amount of the flavor protease is 2kU/g.
5. preparation method according to claim 1, it is characterised in that: the ferrohemoglobin enzymolysis liquid Central Asia obtained by step S3
For Ferri-hemoglobin degree of hydrolysis up to 27.3~34.2%, heme peptide yield is 82.6~87.5%.
6. preparation method according to claim 1, it is characterised in that: in heme oligopeptides concentrate obtained by step S4
The content of heme oligopeptides is 20.8~22.5mg/mL, and the rate of recovery of heme is 83.5~84.1%.
7. preparation method according to claim 1, it is characterised in that: heme oligopeptides microcapsules obtained by step S5
Powder, microcapsule embedded rate are 95.83~97.34%, heme oligopeptides 203~207mg/g of content, heme yield
It is 77.64~79.33%.
8. a kind of heme oligopeptides microcapsule powder being prepared by preparation method described in claim 1.
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