CN101353686A - Production method of haemachrome and decolorizing blood plasma albumen powder - Google Patents

Production method of haemachrome and decolorizing blood plasma albumen powder Download PDF

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Publication number
CN101353686A
CN101353686A CNA200810120963XA CN200810120963A CN101353686A CN 101353686 A CN101353686 A CN 101353686A CN A200810120963X A CNA200810120963X A CN A200810120963XA CN 200810120963 A CN200810120963 A CN 200810120963A CN 101353686 A CN101353686 A CN 101353686A
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protoheme
production method
decolouring
globulin powder
enzymolysis
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CN101353686B (en
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龚兴国
郦剑勇
秦勇
曾冬云
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Danyang Biological Polytron Technologies Inc Of Fushun
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Zhejiang University ZJU
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Abstract

The invention discloses a manufacture method of hemachrome and discolored globin powder. The method comprises the steps that: after zymohydrolysis and equal point deposition of the raw material hemoglobin are carried out, the globin powder is prepared by supernatant by concentration and drying; and the hemachrome is prepared by precipitation and crystallization. The method of the invention can fully utilize the precious protein resources of poultry blood, jointly extract the hemachrome and eatable discolored globin powder and has not waste; the obtained discolored globin powder has good color, few impurities and high content of proteins. The method has the advantages of simple operation, low lost and clean without contamination, and being suitable for industrial production.

Description

The production method of protoheme and decolouring globulin powder
Technical field
The present invention relates to the production method of protoheme and decolouring globulin powder, belong to food chemistry and biochemical field.
Background technology
Blood accounts for the 7%-8% of domestic birds and animals total protein concentration, and itself is nutritious, and protein content is up to about 20%, have " liquid meat " and title.Albumen has albumin, oxyphorase etc. in the blood, and wherein oxyphorase accounts for 70%-80%.But because the disgusting fishy smell of oxyphorase, except being used for blood bean curd on a small quantity, major part is dropped, and has not only wasted the protein resource of a large amount of preciousnesses but also has polluted ecotope.Oxyphorase is a homotetramer of being made up of four subunits, and each subunit contains a globin and one bonded protoheme with it.Wherein a kind of especially good hematinic of protoheme and important medicine chemical material.Therefore making full use of blood resource can protect environment again and can create good economic benefit by efficent use of resources, is subjected to domestic and international extensive concern.
Heme moiety and the globin part of fully rationally utilizing the artis of blood resource to be to separate oxyphorase have proposed a variety of separation methods now.Traditional method is utilized the organic solvent extracting, comprises the Glacial acetic acid extracting, the extracting of small molecules ketone such as acetone, methylethylketone etc., small molecular alcohol extracting such as methyl alcohol, ethanol etc.The organic solvent extracting is the extracting protoheme well, but needs very a large amount of organic solvents, and the protoheme that extracts 1kg needs the organic solvent of 800L-1000L, so inefficiency, brings huge inconvenience to operation, and also bigger to environmental hazard.U.S. Pat 4376727 proposes to adsorb protoheme with carboxymethyl cellulose, this method can solve protoheme and separate with globin, but what obtain is the mixture of protoheme and carboxymethyl cellulose, can not obtain pure protoheme, and carboxymethyl cellulose can not be regenerated, therefore this method cost is higher, has limited its use.Chinese patent ZL87101834 and ZL200610080022.9 use tensio-active agent as catalyzer crystallization protoheme, crystallization effect is better, but this method blood cell is easy to generate globin and protoheme co-precipitation in heat-processed, and address this problem just needs accurately control reaction process and utilization complicated operations equipment; The more important thing is that the protein powder that this method generates contains tensio-active agent, its person poultry safety's property is under suspicion.
Summary of the invention
The purpose of this invention is to provide a kind of simple to operate, with low cost, free of contamination protoheme and the decolouring globulin powder production method.
The production method of protoheme of the present invention and decolouring globulin powder, its step is as follows
1) the oxyphorase raw material is dissolved in water and stirs, the preparation mass concentration is at the 1%-20% haemoglobin aqueous solution, add proteolytic ferment then, the addition of proteolytic ferment is the 0.05%-10% of protein content in the hemoglobin solutions, regulate pH1.5~9.0, carry out enzymolysis under 25~70 ℃ of temperature, obtain enzymolysis solution, the enzymolysis extent control is at degree of hydrolysis 5%~40%;
2) the pH value of regulating enzymolysis solution is filtered to 2.0-6.0, collects the protoheme precipitation, gets the protoheme work in-process after the drying, and filtrate is not for containing the protein solution of protoheme;
3) add entry, concentrated hydrochloric acid and nonionogenic tenside in the protoheme work in-process, and stir, with dry weight basis, control protoheme work in-process: water: concentrated hydrochloric acid: the mass ratio of nonionogenic tenside is 1: 10-200: 0.1-10: 0.2-5, continue to stir, and be heated to 70 ℃~95 ℃, centrifugal, collect the protoheme crystallization, wash after drying with water, get the finished product protoheme;
4) regulating step 2) pH of gained filtrate to neutral, concentrate, dry, globulin powder must decolour.
Among the present invention, said oxyphorase raw material is livestock blood or is oxyphorase, anticoagulated blood, clot, globulin powder or the blood cell of livestock blood processing.
Among the present invention, proteolytic ferment do not had special requirement, as long as the proteolytic enzyme of energy hydrolysis oxyphorase is all available, stomach en-, trypsinase as animal-origin, the papoid of plant origin, bromeline, microbe-derived animal proteolytic enzyme (production of Nanning Pang Bo bio tech ltd), food flavor enzyme or the like can be with any or its mixtures in above-mentioned.Because animal proteolytic enzyme and food flavor enzyme low price, easy to use and commercial production marketing, preferred animal proteolytic enzyme and food flavor enzyme, perhaps both mixed enzyme arranged.
In order to eliminate protease activities, to improve the quality of products, further feature of the present invention is the enzymolysis solution that step 1) obtains to be heated to keep 20 minutes inactivated proteases more than 90 ℃.
Among the present invention, said nonionogenic tenside is a kind of or its mixture among alkylphenol polyoxyethylene, fatty alcohol-polyoxyethylene ether, tween and the Si Pan.Because tween is extensive use of in daily life as foodstuff additive, its security obtains proof already, and preferred tween is as the tensio-active agent of crystallization protoheme.
The mass concentration of used concentrated hydrochloric acid is 35%~38% among the present invention.
The inventive method is before the crystallization protoheme, and the oxyphorase enzymolysis is micromolecular albumen, and thermostability improves, not can with the protoheme co-precipitation; By regulating the pH value of enzymolysis solution, most albumen is separated with protoheme, the protoheme crystallization just than being easier to, is reacted easier operation like this, and whole process does not need special equipment, does not need to add crystal seed yet.
The decolouring protein powder color and luster that the present invention obtains is good, and impurity is few, and protein content reaches about 95%, can be used for foodstuff additive.
The inventive method is simple to operate, with low cost, can utilize the oxyphorase that extracts behind the superoxide-dismutase (SOD) as raw material, unite and extract protoheme and food grade decolouring globulin powder, thereby realized the comprehensive utilization of livestock blood, do not have waste, cleanliness without any pollution is applicable to suitability for industrialized production.
Embodiment
Embodiment 1
Fresh pig blood 300L is collected in the slaughterhouse, adds the anti-freezing of 1kg Trisodium Citrate, after adding 300L water and stirring, adds the 1kg animal proteolytic enzyme, and the 1kg food flavor enzyme is regulated pH value to 7.5, is heated to 55 ℃, enzymolysis 48 hours.Heating enzymolysis solution to 90 ℃ insulation 20min inactivated proteases.Regulate the pH to 6.0 of enzymolysis solution after the cooling with hydrochloric acid, centrifugal then supernatant liquor and the precipitation of collecting respectively.Supernatant liquor is slightly yellow, regulates pH to 7 with sodium hydroxide solution, and triple effect concentrates, and spraying drying gets white protein powder 50kg; Precipitation (containing dry-matter 10kg approximately) adds 50L water and stirs, add the 1L mass concentration and be 38% concentrated hydrochloric acid, the 50kg polysorbas20, continue to stir, be heated to 70 ℃ then and stop to stir, insulation 30min, cooling back centrifugal collecting precipitation, dry after this precipitation washes with water finished product protoheme 1.5kg, purity is 91.3%, iron recovery is 91%.
Embodiment 2
Fresh ox blood 300L is collected in the slaughterhouse, adds the potassium oxalate anti-freezing, continuous centrifuge separate blood plasma 170L, blood cell 130L.The blood plasma spraying drying makes plasma protein powder 13kg.Add 200L water in the blood cell, stir 30 minutes haemolysis, regulate pH value to 9.0 with the NaOH solution of 1M, add the 4kg animal proteolytic enzyme, be heated to 65 ℃, enzymolysis to degree of hydrolysis reaches 40%.Enzymolysis solution boiled 30 minutes after enzymolysis was finished, and pH to 4.5 back press filtration is regulated with hydrochloric acid in the cooling back, gets filtrate 280kg, filter residue 50kg (about dry-matter 8kg).Filtrate is regulated pH to 6.5 with sodium hydroxide solution, ultrafiltration and concentration, and spraying drying makes protein powder 40kg.Filter residue adds water 1600L and stirs, add the 80L mass concentration and be 35% concentrated hydrochloric acid, 1.6kg tween 80, stir, being heated to 95 ℃ promptly has the protoheme crystal to separate out, to be crystallized fully after, centrifugal collection protoheme precipitation, protoheme precipitation washes with water, dry finished product protoheme 1.6kg, purity 90.3%.
Embodiment 3
Slaughterhouse chicken and duck blood condenses into clot, gets 500kg, and pulverizer adds 1000L water after pulverizing, and stirs, and regulates pH value to 7.0, adds the 60g animal proteolytic enzyme, is heated to 70 ℃, and enzymolysis 12 hours, degree of hydrolysis are 5.6%.Behind hydrochloric acid adjusting pH to 2.5, centrifugal clear liquid and the precipitation of collecting respectively, supernatant liquor is regulated pH to 5.5 with sodium hydroxide solution, and triple effect concentrates, and spraying drying gets white protein powder 105kg; Precipitation adds water 800L and stirs, and adds the 100L mass concentration and be 36% concentrated hydrochloric acid, 4kg polysorbas20,1kg Si Pan 60, stirs, be heated to 75 ℃ and stop to stir, 15min is left standstill in insulation, cooling, centrifugal collecting precipitation, precipitation dry after washing with water finished product protoheme 2.7kg, purity 90%.
Embodiment 4
Globulin powder 30kg adds 2500L water, stirs, and regulates pH value to 1.5, adds the 150g stomach en-, is heated to 37 ℃, enzymolysis 72 hours.Regulate pH to 6.0 with sodium hydroxide after enzymolysis is finished, press filtration gets filtrate 2400L.The filtrate triple effect concentrates, and spraying drying gets protein powder 26kg; Filter residue (the about 5kg of dry weight) adds water 500L and stirs, add the 40L mass concentration and be 35% concentrated hydrochloric acid, 25kg fatty alcohol-polyoxyethylene ether paregal O 15, stir, be heated to 88 ℃ and stop to stir, insulation 45min, the cooling back is centrifugal, collecting precipitation also washes with water, dry finished product protoheme 1.1kg, the purity 88% of getting.
Embodiment 5
Commercially available blood meal 200kg adds 800L water, stirs, regulate pH value to 7.8, add the 10kg food flavor enzyme, 25 ℃ of enzymolysis 12 hours, this moment, degree of hydrolysis was 5.3%, boil the 30min inactivated proteases, regulate pH4.5 with phosphoric acid, press filtration is collected filtrate respectively with filter residue, and filtrate is regulated pH to 7.0 with sodium hydrogen carbonate solution, ultrafiltration and concentration, spraying drying get white protein powder 160kg; Filter residue (the about 40kg of dry matter content) adds water 500L and stirs, add the 40L mass concentration and be 35% concentrated hydrochloric acid, the 50kg polysorbas20, the 50kg tween stirs, being heated to 95 ℃ stops to stir, insulation 5min, cooling back centrifugal collecting precipitation is after this precipitation washes with water, oven dry is finished product protoheme 5kg, purity 92%.
Embodiment 6
Anti-freezing pig blood 300L adds 300L water, stirs, and with salt acid for adjusting pH value to 5.7, adds the 1kg papoid, is heated to 58 ℃, enzymolysis 48 hours.The enzymolysis post-heating boils inactivated proteases, centrifugal supernatant liquor and the precipitation of collecting respectively in cooling back.Supernatant liquor is slightly yellow, regulates pH to 7 with sodium hydroxide solution, and triple effect concentrates, and spraying drying gets white protein powder 50kg; Precipitation adds 50L water and stirs, add the 4L mass concentration and be 35% concentrated hydrochloric acid, 10kg polyoxyethylene nonylphenol ether NP-10, continue to stir, be heated to 70 ℃ then and stop to stir, insulation 30min, cooling back centrifugal collecting precipitation, dry after this precipitation washes with water finished product protoheme 1.6kg, purity is 90%.
Embodiment 7
Anti-freezing pig blood 300L, the centrifugal blood cell 130L that gets, blood plasma 170L, the blood plasma spraying drying is made plasma protein powder 13kg, adds water 600L in the blood cell, after stirring 30min, add an amount of sodium-chlor, and with salt acid for adjusting pH value to 5.0,60 ℃ of insulation 30min, filter, the further processing of supernatant liquor can obtain superoxide-dismutase.Filter residue mainly is an oxyphorase, adds 600L water in the filter residue, regulates pH value to 7.0, adds 1kg conjugated protein lytic enzyme, and the 1kg food flavor enzyme is heated to 58 ℃, insulation enzymolysis 48 hours.Enzymolysis is finished post-heating and is boiled inactivated proteases, centrifugal supernatant liquor and the precipitation of collecting respectively in cooling back.Supernatant liquor is slightly yellow, and triple effect concentrates, and spraying drying gets white protein powder 45kg; Precipitation (containing dry-matter 10kg approximately) adds 50L water and stirs, add the 4L mass concentration and be 35% concentrated hydrochloric acid, 10kg polyoxyethylene nonylphenol ether NP-10, continue to stir, be heated to 70 ℃ then and stop to stir, insulation 60min, cooling back centrifugal collecting precipitation, dry after this precipitation washes with water finished product protoheme 1.5kg, purity is 91%.

Claims (5)

1, the production method of a kind of protoheme and decolouring globulin powder, its step is as follows:
1) the oxyphorase raw material is dissolved in water and stirs, the preparation mass concentration is at the 1%-20% haemoglobin aqueous solution, add proteolytic ferment then, the addition of proteolytic ferment is the 0.05%-10% of protein content in the hemoglobin solutions, regulate pH1.5~9.0, carry out enzymolysis under 25~70 ℃ of temperature, obtain enzymolysis solution, the enzymolysis extent control is at degree of hydrolysis 5%~40%;
2) the pH value of regulating enzymolysis solution is filtered to 2.0-6.0, collects the protoheme precipitation, gets the protoheme work in-process after the drying, and filtrate is not for containing the protein solution of protoheme;
3) add entry, concentrated hydrochloric acid and nonionogenic tenside in the protoheme work in-process, and stir, with dry weight basis, control protoheme work in-process: water: concentrated hydrochloric acid: the mass ratio of nonionogenic tenside is 1: 10-200: 0.1-10: 0.2-5, continue to stir, and be heated to 70 ℃~95 ℃, centrifugal, collect the protoheme crystallization, wash after drying with water, get the finished product protoheme;
4) regulating step 2) pH of gained filtrate to neutral, concentrate, dry, globulin powder must decolour.
2, the production method of protoheme according to claim 1 and decolouring globulin powder is characterized in that said oxyphorase raw material is livestock blood or oxyphorase, anticoagulated blood, clot, globulin powder or the blood cell processed for livestock blood.
3, the production method of protoheme according to claim 1 and decolouring globulin powder is characterized in that said proteolytic ferment is a kind of or its mixture in stomach en-, trypsinase, papoid, bromeline, animal proteolytic enzyme and the food flavor enzyme.
4, according to the production method of described protoheme of claim 1 and decolouring globulin powder, it is characterized in that said nonionogenic tenside is a kind of or its mixture among alkylphenol polyoxyethylene, fatty alcohol-polyoxyethylene ether, tween and the Si Pan.
5,, it is characterized in that the enzymolysis solution that step 1) obtains is heated to 20 minutes inactivated proteases of maintenance more than 90 ℃ according to the production method of described protoheme of claim 1 and decolouring globulin powder.
CN200810120963XA 2008-09-09 2008-09-09 Production method of haemachrome and decolorizing blood plasma albumen powder Expired - Fee Related CN101353686B (en)

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Cited By (14)

* Cited by examiner, † Cited by third party
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CN102077901A (en) * 2010-12-14 2011-06-01 成都宏安生物科技有限公司 Swine hemoglobin enzymolysis and decoloration method
CN102125164A (en) * 2010-10-29 2011-07-20 成都天屹生物科技有限公司 Preparation method of pig blood fibrin polypeptide powder
CN103396418A (en) * 2013-07-23 2013-11-20 甘肃农业大学 Enzymatic preparation method of hemin by using dried yak blood as raw material
CN104256199A (en) * 2014-10-16 2015-01-07 浙江索纳克生物科技有限公司 Loach feed added with porcine hemoglobin powder
CN104256198A (en) * 2014-10-16 2015-01-07 浙江索纳克生物科技有限公司 Soft-shelled turtle feed with spray-dried pig blood cells added
CN105385735A (en) * 2015-11-25 2016-03-09 重庆三零三科技有限公司 Preparation method of chicken blood antimicrobial peptide
CN106107574A (en) * 2016-08-11 2016-11-16 安徽省农业科学院农产品加工研究所 A kind of preparation method of Sanguis sus domestica source compound microelement supplement
CN106261817A (en) * 2016-08-11 2017-01-04 安徽省农业科学院农产品加工研究所 A kind of efficient enzymolysis Application way of Sanguis sus domestica
CN106261816A (en) * 2016-08-11 2017-01-04 安徽省农业科学院农产品加工研究所 A kind of Sanguis sus domestica source iron supplementary intermediate and the preparation method of finished product thereof
CN106554979A (en) * 2016-11-18 2017-04-05 安徽菁硕科技有限公司 A kind of method of the production haemachrome that combined with acid acetone extraction by enzymatic isolation method
CN110183497A (en) * 2019-07-17 2019-08-30 甘肃养泰和生物科技有限公司 A method of purifying hemin from animal blood cell
CN114592029A (en) * 2022-03-09 2022-06-07 山西勰成生物科技有限公司 Preparation method of fermented enzymatic decolorized hemoglobin
CN115386000A (en) * 2022-09-14 2022-11-25 江苏省农业科学院 Method for preparing decolorized peptide and heme peptide from poultry blood cell processing byproducts
CN117886826A (en) * 2023-12-13 2024-04-16 甘肃养泰和生物科技有限公司 Method for extracting methemoglobin by heating and pressurizing

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CN1188408C (en) * 2000-12-28 2005-02-09 内蒙古草原兴发股份有限公司 Process for extracting heme from animal blood
CN100390175C (en) * 2003-04-11 2008-05-28 王利忠 Process for extracting and puritying haem
CN100378103C (en) * 2006-04-28 2008-04-02 新疆科丽生物技术有限公司 Process for extracting protohemin by using surfactant
CN100581377C (en) * 2007-11-30 2010-01-20 华南理工大学 Apocytochrome porcine hemoglobin hydrolyzates and protoheme peptide preparing method

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102125164A (en) * 2010-10-29 2011-07-20 成都天屹生物科技有限公司 Preparation method of pig blood fibrin polypeptide powder
CN102125164B (en) * 2010-10-29 2013-08-28 成都天屹生物科技有限公司 Preparation method of pig blood fibrin polypeptide powder
CN102077901A (en) * 2010-12-14 2011-06-01 成都宏安生物科技有限公司 Swine hemoglobin enzymolysis and decoloration method
CN102077901B (en) * 2010-12-14 2013-04-24 成都宏安生物科技有限公司 Swine hemoglobin enzymolysis and decoloration method
CN103396418A (en) * 2013-07-23 2013-11-20 甘肃农业大学 Enzymatic preparation method of hemin by using dried yak blood as raw material
CN104256199A (en) * 2014-10-16 2015-01-07 浙江索纳克生物科技有限公司 Loach feed added with porcine hemoglobin powder
CN104256198A (en) * 2014-10-16 2015-01-07 浙江索纳克生物科技有限公司 Soft-shelled turtle feed with spray-dried pig blood cells added
CN105385735A (en) * 2015-11-25 2016-03-09 重庆三零三科技有限公司 Preparation method of chicken blood antimicrobial peptide
CN106107574A (en) * 2016-08-11 2016-11-16 安徽省农业科学院农产品加工研究所 A kind of preparation method of Sanguis sus domestica source compound microelement supplement
CN106261817A (en) * 2016-08-11 2017-01-04 安徽省农业科学院农产品加工研究所 A kind of efficient enzymolysis Application way of Sanguis sus domestica
CN106261816A (en) * 2016-08-11 2017-01-04 安徽省农业科学院农产品加工研究所 A kind of Sanguis sus domestica source iron supplementary intermediate and the preparation method of finished product thereof
CN106554979A (en) * 2016-11-18 2017-04-05 安徽菁硕科技有限公司 A kind of method of the production haemachrome that combined with acid acetone extraction by enzymatic isolation method
CN110183497A (en) * 2019-07-17 2019-08-30 甘肃养泰和生物科技有限公司 A method of purifying hemin from animal blood cell
CN114592029A (en) * 2022-03-09 2022-06-07 山西勰成生物科技有限公司 Preparation method of fermented enzymatic decolorized hemoglobin
CN115386000A (en) * 2022-09-14 2022-11-25 江苏省农业科学院 Method for preparing decolorized peptide and heme peptide from poultry blood cell processing byproducts
CN117886826A (en) * 2023-12-13 2024-04-16 甘肃养泰和生物科技有限公司 Method for extracting methemoglobin by heating and pressurizing

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