CN103255190A - Method and process for extracting animal micro-molecule polypeptides and amino acids by utilizing complex enzyme - Google Patents

Method and process for extracting animal micro-molecule polypeptides and amino acids by utilizing complex enzyme Download PDF

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Publication number
CN103255190A
CN103255190A CN2013102065877A CN201310206587A CN103255190A CN 103255190 A CN103255190 A CN 103255190A CN 2013102065877 A CN2013102065877 A CN 2013102065877A CN 201310206587 A CN201310206587 A CN 201310206587A CN 103255190 A CN103255190 A CN 103255190A
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Prior art keywords
enzymolysis
animal
prozyme
enzyme
temperature
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CN2013102065877A
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于国友
王佃亮
于丽萍
王思远
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GENERAL HOSPITAL OF SECOND ARTILLERY OF CHINESE PLA
WEIHAI KANGBOER BIOLOG PHARMACEUTICAL CO Ltd
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GENERAL HOSPITAL OF SECOND ARTILLERY OF CHINESE PLA
WEIHAI KANGBOER BIOLOG PHARMACEUTICAL CO Ltd
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Priority to CN2013102065877A priority Critical patent/CN103255190A/en
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Abstract

The invention discloses a method and process for extracting animal micro-molecule polypeptides and amino acids by utilizing a complex enzyme. The method comprises the following steps: (1) crushing animal tissues so that the animal tissues can pass through a sieve of 80-200 meshes, and adding water to prepare a suspension with the concentration of 8-30 percent; (2) raising the temperature to be 35-40 DEG C, regulating the pH value to be 8.0-9.0 by using sodium carbonate, adding a complex enzyme I accounting for 0.5-2.0 percent of the animal tissues, performing ultrasonic enzymolysis at 14-16 kHz for 1-2 hours so that the proteins between cells are subjected to full enzymolysis, regulating the pH value to be 6.5-7.5, adding a complex enzyme II accounting for 0.5-3.0 percent of the animal tissues, fully and uniformly stirring, controlling the temperature of the water bath to be 37-45 DEG C, and performing ultrasonic enzymolysis at 15-17 kHz for 1-6 hours; (3) raising the temperature of the enzymolysis solution to be 90-100 DEG C to inactivate the enzyme for 20-30 minutes after the enzymolysis is finished; (4) filtering the solution subjected to enzymolysis through a 0.22mu m microfiltration membrane; and (5) concentrating the filtrate to the relative density of 1:1.0-1:1.2 to prepare the concentrated solution, wherein the complex enzyme II consists of Alcalase, Flavourzyme, Protamex, neutral protease and papain. According to the method, the enzymolysis efficiency can be improved, and the production cost can be reduced.

Description

Utilize prozyme to extract animal micromolecule polypeptide and amino acid whose method and technology
Technical field
The invention belongs to the protein engineering field, particularly relate to a kind of prozyme that utilizes and extract animal micromolecule polypeptide and amino acid whose method and technology.
Background technology
Compare with plant, the composition of the range protein in the animal body and ratio and people are more approaching, thereby the protein component in the animal body has higher nutritive value to the people, especially some have micromolecule polypeptide and the amino acid (biologically active substance) of particular physiological function in the animal body, and metabolism has important regulatory role to human body.Traditional animal series products mainly adopts physical method (as fragmentation, high temperature etc.) to process, and not only extraction efficiency is low, also can cause the mass efficient composition to destroy loss of biological activity.Although now indivedual courses of processing have adopted biological enzymolysis technology, seriously ignored a problem (choosing of enzyme should be complementary with animal tissues's structure), make the degradation effect of enzyme not ideal enough.Enzyme is produced by the living organisms cell, is subjected to multiple factor to regulate the biological catalyst with catalytic capability of control.Compared its common point with general catalyzer, but outstanding feature (as: the catalytic efficiency height of enzyme has height specificity etc.) has been arranged.The kind of enzyme is a lot, and what be used for the animal enzymolysis mainly is proteolytic enzyme.Proteolytic enzyme is divided into endopeptidase and exopeptidase two classes again, want to take full advantage of the effective constituent in the animal body, just need endopeptidase and exopeptidase combination, with micromolecule polypeptide and the amino acid that abundant degrade proteins macromole utilizes for ease of absorption of human body, this just needs a kind of complex enzyme hydrolysis technology of invention for the protein degradation in the animal body.
Summary of the invention
The invention provides a kind of prozyme that utilizes and extract animal micromolecule polypeptide and amino acid whose method and technology, simple to operate, production cost is low, has improved the utilization ratio of animal protein nutritive ingredient.
The technical solution adopted in the present invention is:
Prozyme is used for extracting animal micromolecule polypeptide and amino acid whose method and concrete production technique:
(1) animal tissues is pulverized, can pass through 80 orders~200 mesh sieve, adding water, to be made into concentration be 8%~30% suspension;
(2) improving temperature is 35 ℃~40 ℃, regulating the pH value with yellow soda ash is 8.0~9.0, the prozyme I that adds animal tissues 0.5%~2.0% again, place it in the Vltrasonic device then, the ultrasonic enzymolysis 1h~2h of 14~16kHz, make the abundant enzymolysis of intercellular albumen, regulating the pH value with yellow soda ash again is 6.5~7.5, add the compound enzyme II that is equivalent to animal tissues 0.5%~3.0%, fully stir evenly, the temperature of control water-bath is at 37 ℃~45 ℃, the ultrasonic enzymolysis 1h~6h of 15~17kHz, in enzymolysis, need suitable stirring water-bath, be conducive to the hydrolysis effect of enzyme like this, shorten enzymolysis time;
(3) after enzymolysis is finished, improve enzymolysis solution temperature to 90 ℃~100 ℃, inactivator 20~30min;
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane;
(5) getting filtrate, to be concentrated into relative density be 1: 1.0~1: 1.2, makes concentrated solution.
Prozyme comprises prozyme I and compound enzyme II.
Prozyme I mainly is made up of trypsinase 30%~70%, steapsase 10%~40% and pancreatic amylase 5%~30%, and the activity unit of enzyme is 20,000 U/g~200,000 U/g, can decompose the protein of intercellular fat and part.
Compound enzyme II is made up of Alcalase (Sumizyme MP) 5%~30%, Flavourzyme (compound protease) 5%~30%, Protamex (compound protease) 5%~30%, neutral protease 10%~50%, papoid 1%~30%, total mass consists of 100%, and the activity unit of enzyme is 50,000 U/g~300,000 U/g.
Advantage of the present invention: utilize enzyme reaction to have highly narrow spectrum characteristics, select endopeptidase and exopeptidase to be mixed in proportion, high molecular weight protein in the animal body is decomposed into micromolecule polypeptide and the amino acid that is conducive to absorption of human body, destruction by cell membrane discharges intracellular effective constituent, can improve enzymolysis efficiency, can reduce production costs again.Can be applicable to the processing of bright sea cucumber, abalone, oyster, pig bone, beef, chicken gizzard, fish-bone, pig's feet, shank, pigskin, ox-hide.
Embodiment
Following examples are used for understanding the present invention better, but are not used for limiting the scope that the present invention protects.
Embodiment 1: utilize prozyme that sea cucumber is carried out enzymolysis.
(1) get bright sea cucumber 1kg, pulverizing can be crossed 80 mesh sieve.
(2) add water and make animal tissues's suspension of 10%, 100 ℃ of heating 30min when temperature is down to 35 ℃~40 ℃, are 8.0 with the adjusting PH with base value, add enteropeptidase, the back adding that stirs is equivalent to the prozyme I of animal tissues 1.0%, and 37 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 1h of 15kHz, adjust pH is 7.0, add the compound enzyme II that is equivalent to animal tissues 1.5%, 45 ℃ of control temperature, the ultrasonic enzymolysis 2h of 16.5kHz.
(3) improve temperature to 100 ℃, quick inactivating enzyme 30min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.0, makes tan concentrated solution.
Embodiment 2: utilize prozyme that bright Oyster is carried out enzymolysis.
(1) get Oyster 1kg, pulverizing can be crossed 100 mesh sieve.
(2) add water and make 25% suspension, improving temperature is 35 ℃~40 ℃, be 8.5 with the yellow soda ash adjust pH, the back adding that stirs is equivalent to the Cotazym I of animal tissue contg 0.8%, and 40 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 1.5h of 14.5kHz, adjust pH is 7.5, add the compound enzyme II that is equivalent to animal tissue contg 1.0%, 40 ℃ of control temperature, the ultrasonic enzymolysis 4h of 15kHz.
(3) improve temperature to 100 ℃, quick inactivating enzyme 20min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.0, makes concentrated solution.
Embodiment 3: utilize prozyme that abalone is carried out enzymolysis.
(1) get abalone meat 1kg, pulverizing can be crossed 100 mesh sieve.
(2) add water and make 20% suspension, when the raising temperature is 35 ℃~40 ℃, be 8.5 with the sodium bicarbonate adjust pH, the back adding that stirs is equivalent to the prozyme I of animal tissue contg 1.1%, and 35 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 1.0h of 16kHz, adjust pH is 6.5, add the compound enzyme II that is equivalent to animal tissue contg 1.2%, temperature is controlled at 37 ℃, the ultrasonic enzymolysis 2h of 17kHz.
(3) improve temperature to 95 ℃, quick inactivating enzyme 30min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.0, makes concentrated solution.
Embodiment 4: utilize prozyme that the pig bone is carried out enzymolysis.
(1) get pig bone 2kg, pulverizing can be crossed 100 mesh sieve.
(2) add water and make 15% suspension, when the raising temperature is 35 ℃~40 ℃, be 8.5 with the yellow soda ash adjust pH, the back adding that stirs is equivalent to the prozyme I of animal tissue contg 1.5%, and 36 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 2.0h of 16kHz, adjust pH is 6.5, add the compound enzyme II that is equivalent to animal tissue contg 2.0%, 42 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 3h of 17kHz.
(3) improve temperature to 90 ℃, quick inactivating enzyme 30min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.2, makes concentrated solution.
Embodiment 5: utilize prozyme that chicken gizzard is carried out enzymolysis.
(1) get chicken gizzard 1kg, pulverizing can be crossed 90 mesh sieve.
(2) add water and make 8% suspension, 80 ℃ of heating 30min, when reducing the temperature to 35 ℃~40 ℃, be 8.0 with the yellow soda ash adjust pH, the back that stirs adds the prozyme I that is equivalent to animal tissue contg 1.2%, 38 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 2.0h of 15kHz, adjust pH is 7.2, adds the compound enzyme II that is equivalent to animal tissue contg 2.5%, 39 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 2h of 16kHz.
(3) improve temperature to 90 ℃, quick inactivating enzyme 30min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.0, makes concentrated solution.
Embodiment 6: utilize prozyme that fish-bone is carried out enzymolysis.
(1) get fish-bone 1kg, pulverizing can be crossed 100 mesh sieves.
(2) add water and make 30% suspension, during rising temperature to 35 ℃~40 ℃, be 8.5 with the adjusting PH with base value, the back adding that stirs is equivalent to the prozyme I of animal tissue contg 1.0%, and 37 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 2.0h of 15kHz, adjust pH is 6.5, add the compound enzyme II that is equivalent to animal tissue contg 0.5%, 55 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 5h of 17kHz.
(3) improve temperature to 100 ℃, quick inactivating enzyme 25min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.1, makes concentrated solution.
Embodiment 7: utilize prozyme that beef is carried out enzymolysis.
(1) get beef 1kg, pulverizing can be crossed 80 mesh sieves.
(2) add water and make 20% suspension, when improving temperature to 35 ℃~40 ℃, be 9.0 with the yellow soda ash adjust pH, the back adding that stirs is equivalent to the prozyme I of animal tissue contg 0.5%, and 35 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 1.5h of 14kHz, adjust pH is 7.0, add the compound enzyme II that is equivalent to animal tissue contg 3.0%, 45 ℃ of constant temperature are hatched, the ultrasonic enzymolysis 3h of 15kHz.
(3) improve temperature to 100 ℃, quick inactivating enzyme 20min, naturally cooling cooling then.
(4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane.
(5) getting filtrate, to be concentrated into relative density be 1: 1.05, makes concentrated solution.

Claims (4)

1. utilize prozyme to extract animal micromolecule polypeptide and amino acid whose method and technology, it is characterized in that prozyme is used to extract animal micromolecule polypeptide and amino acid whose method and concrete production technique and is: (1) pulverizes animal tissues, can pass through 80 orders~200 mesh sieve, adding water, to be made into concentration be 8%~30% suspension; (2) improving temperature is 35 ℃~40 ℃, regulating the pH value with yellow soda ash is 8.0~9.0, the prozyme I that adds animal tissues 0.5%~2.0% again, place it in the Vltrasonic device then, the ultrasonic enzymolysis 1h~2h of 14~16kHz, make the abundant enzymolysis of intercellular albumen, regulating the pH value with yellow soda ash again is 6.5~7.5, add the compound enzyme II that is equivalent to animal tissues 0.5%~3.0%, fully stir evenly, the temperature of control water-bath is at 37 ℃~45 ℃, the ultrasonic enzymolysis 1h~6h of 15~17kHz, in enzymolysis, need suitable stirring water-bath, be conducive to the hydrolysis effect of enzyme like this, shorten enzymolysis time; (3) after enzymolysis is finished, improve enzymolysis solution temperature to 90 ℃~100 ℃, inactivator 20~30min; (4) with the solution behind the enzymolysis through 0.22 μ m filtering with microporous membrane; (5) getting filtrate, to be concentrated into relative density be 1: 1.0~1: 1.2, makes concentrated solution.
2. the prozyme that utilizes according to claim 1 extracts animal micromolecule polypeptide and amino acid whose method and technology, it is characterized in that prozyme I mainly is made up of trypsinase 30%~60%, steapsase 10%~30% and pancreatic amylase 5%~20%, the activity unit of enzyme is 20,000 U/g~200,000 U/g, can decompose the protein of intercellular fat and part.
3. the prozyme that utilizes according to claim 1 extracts animal micromolecule polypeptide and amino acid whose method and technology, it is characterized in that compound enzyme II is made up of Sumizyme MP 5%~30%, compound protease 5%~30%, compound protease 5%~30%, neutral protease 10%~50%, papoid 1%~30%, total mass consists of 100%, and the activity unit of enzyme is 50,000 U/g~300,000 U/g.
4. the prozyme that utilizes according to claim 1 extracts animal micromolecule polypeptide and amino acid whose method and technology, it is characterized in that can be applicable to the processing of bright sea cucumber, abalone, oyster, pig bone, beef, chicken gizzard, fish-bone, pig's feet, shank, pigskin, ox-hide.
CN2013102065877A 2013-05-30 2013-05-30 Method and process for extracting animal micro-molecule polypeptides and amino acids by utilizing complex enzyme Pending CN103255190A (en)

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CN103553749A (en) * 2013-11-11 2014-02-05 李世泰 Method for producing amino acid agricultural fertilizer through compound enzyme hydrolysis of fish skins and fish scales
CN103932267A (en) * 2014-04-30 2014-07-23 徐海菊 Processing technique for preparing oyster meat nutritious powder by enzymolysis with assistance of ultrasound
CN103947819A (en) * 2014-04-30 2014-07-30 桂林军供生化技术开发有限公司 Method for extracting protein from pigskin
CN104068423A (en) * 2014-06-12 2014-10-01 渤海大学 Oyster meat nutrition powder and preparation method thereof
CN104621338A (en) * 2015-03-03 2015-05-20 浙江小二黑食品有限公司 Method for preparing novel silkie bone polypeptides
CN104805164A (en) * 2015-04-30 2015-07-29 中国食品发酵工业研究院 Preparation method of high protein oyster active peptides with low sensitization
CN104894198A (en) * 2015-04-30 2015-09-09 中国食品发酵工业研究院 Preparation method of hypoallergenic total-nutrient oyster active peptide
CN104974223A (en) * 2015-05-12 2015-10-14 舟山拓智科技服务有限公司 Actinopyga mauritiana polypeptides and application thereof
CN105107815A (en) * 2015-06-25 2015-12-02 河南农业大学 Method for treating illness-dead livestock and poultry by combining chemical method with biological method
CN105200105A (en) * 2015-09-10 2015-12-30 烟台源力德海洋生物有限公司 Preparation method of oyster protein liver protecting peptide preparation
CN106922954A (en) * 2015-12-31 2017-07-07 武汉新华扬生物股份有限公司 A kind of method for preparing chicken feet hide collagen raw material
CN107467344A (en) * 2017-09-20 2017-12-15 杨智 The biologically active peptide and its extracting method extracted in a kind of Phasianidae animal from Galliformes
CN107475339A (en) * 2017-09-20 2017-12-15 广东雅道生物科技有限公司 A kind of meat polypeptide extracting method
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CN107821727A (en) * 2017-10-31 2018-03-23 宁波海博士特医食品科技有限公司 A kind of processing method of sea cucumber Gly-His-Lys
CN107912768A (en) * 2017-12-22 2018-04-17 东阿阿胶股份有限公司 One breeding ass intacellin and preparation method thereof
CN108486087A (en) * 2018-04-10 2018-09-04 吴广兵 It is a kind of enzymolysis chicken gizzard complex enzyme formulation and its application
CN108771244A (en) * 2018-05-24 2018-11-09 南京中生生物科技有限公司 The preparation method of the oligomeric peptide extract of abalone, method, abalone oligopeptide and its application for preparing abalone oligopeptide and abalone powder
CN108977424A (en) * 2018-08-02 2018-12-11 山东隆科特酶制剂有限公司 A kind of catabolic enzyme preparation of the molten slurry of anchovy fish and its application
CN109022400A (en) * 2018-08-20 2018-12-18 山东隆科特酶制剂有限公司 A kind of chicken gizzard degradation complex enzyme preparation special and application
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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
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CN111850064A (en) * 2020-07-28 2020-10-30 宁夏顺宝现代农业股份有限公司 Method for preparing amino acid aqueous solution by using dead chicken
WO2020224058A1 (en) * 2019-05-09 2020-11-12 烟台嘉惠海洋生物科技有限公司 Industrialized production method for preparing oyster peptide by means of enzymatic method
CN112573961A (en) * 2020-12-26 2021-03-30 南宁东恒华道生物科技有限责任公司 Liquid micro-peptide fertilizer produced by composite enzymolysis of urban fresh garbage, production process and application thereof
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CN103553749A (en) * 2013-11-11 2014-02-05 李世泰 Method for producing amino acid agricultural fertilizer through compound enzyme hydrolysis of fish skins and fish scales
CN103553749B (en) * 2013-11-11 2015-01-14 李世泰 Method for producing amino acid agricultural fertilizer through compound enzyme hydrolysis of fish skins and fish scales
CN103932267A (en) * 2014-04-30 2014-07-23 徐海菊 Processing technique for preparing oyster meat nutritious powder by enzymolysis with assistance of ultrasound
CN103947819A (en) * 2014-04-30 2014-07-30 桂林军供生化技术开发有限公司 Method for extracting protein from pigskin
CN104068423A (en) * 2014-06-12 2014-10-01 渤海大学 Oyster meat nutrition powder and preparation method thereof
CN104621338A (en) * 2015-03-03 2015-05-20 浙江小二黑食品有限公司 Method for preparing novel silkie bone polypeptides
CN104805164A (en) * 2015-04-30 2015-07-29 中国食品发酵工业研究院 Preparation method of high protein oyster active peptides with low sensitization
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CN105107815A (en) * 2015-06-25 2015-12-02 河南农业大学 Method for treating illness-dead livestock and poultry by combining chemical method with biological method
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CN109527552A (en) * 2018-12-21 2019-03-29 江苏食品药品职业技术学院 Nutritious food of mid-aged population and preparation method thereof
CN109468357A (en) * 2018-12-25 2019-03-15 河北肽都生物科技有限公司 A kind of preparation method of spleen aminopeptide
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
WO2020224058A1 (en) * 2019-05-09 2020-11-12 烟台嘉惠海洋生物科技有限公司 Industrialized production method for preparing oyster peptide by means of enzymatic method
CN110477402A (en) * 2019-08-09 2019-11-22 质每堂(武汉)健康科技有限公司 Fish peptide, fish-bone peptide, sea cucumber peptide and ginseng peptide compound nutritional product and preparation method thereof
CN111850064A (en) * 2020-07-28 2020-10-30 宁夏顺宝现代农业股份有限公司 Method for preparing amino acid aqueous solution by using dead chicken
CN112573961A (en) * 2020-12-26 2021-03-30 南宁东恒华道生物科技有限责任公司 Liquid micro-peptide fertilizer produced by composite enzymolysis of urban fresh garbage, production process and application thereof
CN113349308A (en) * 2021-05-10 2021-09-07 广州清科生物技术有限公司 Composite zymolyte for pet food and preparation process thereof

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Application publication date: 20130821