CN1291032C - Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack - Google Patents
Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack Download PDFInfo
- Publication number
- CN1291032C CN1291032C CNB2004100241841A CN200410024184A CN1291032C CN 1291032 C CN1291032 C CN 1291032C CN B2004100241841 A CNB2004100241841 A CN B2004100241841A CN 200410024184 A CN200410024184 A CN 200410024184A CN 1291032 C CN1291032 C CN 1291032C
- Authority
- CN
- China
- Prior art keywords
- protein
- oligopeptide powder
- enzymolysis
- protein oligopeptide
- protease
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The present invention relates to an enzymolysis technologic method for preparing protein oligopeptide powder by using the Pacific Ocean alaska pollack. The operating technologic method comprises that the Pacific Ocean pollock fish is minced, and 20 AU/kg to 80 AU/kg hay bacillus endoprotease and 8000 LAPU/kg to 40000 LAPU/kg of aspergillus exterior contact proteinase are added, or 20 AU/kg to 80 AU/kg of bacillus compound proteinase is added, and the bacillus compound proteinase and water are uniformly mixed with a ratio of 1.2/1 to 1/1.5. The mixture is put in an enzyme reactor for zymolysis for 2 to 12 hours, the zymolysis temperature is from 50 DEG C to 60 DEG C, and the initial pH is 6.6. Zymolysis liquid inactivated, centrifugalized, separated by membrane, dried in a spray mode or baked. Then, water solubility protein oligopeptide powder and a side product can be obtained. The protein oligopeptide powder prepared by the present invention is light cream white, has no bitterness and has a little fishy odor. The molecular weight (greater than 85%) can be respectively controlled less than or equal to 1000 Da or from 1000 to 5000Da, and the recovery rate (relatively to the total protein content of raw materials) of the protein oligopeptide powder can reach from 65% to 79%.
Description
Technical field:
The invention belongs to marine biotechnology enzymatic production process technology, is the processing method of a kind of enzymatic hydrolysis Pacific Ocean wall pollack flesh of fish preparation protein oligopeptide powder.
Background technology:
Protease hydrolyzed is the most promising processing methodes of preparation protein oligopeptide goods, economic quality indexs such as the rate of recovery of protein oligopeptide goods, molecular weight (closely related with biological activity), taste local flavor depend primarily on conditions such as selected protease preparation kind, raw material variety, enzymolysis parameter (E/S is enzyme weight or enzyme activity unit/kg protein, material-water ratio, time, temperature both), and the result can present very big-difference.Before the present invention makes, France F.Guerard etc. (2001,2002) adopt Alcalase proteolytic enzyme and " Umamizyme " neutral protease (E/S=1g/kg～15g/kg, material-water ratio 1: 1.5) hydrolysis yellowfin tuna byproduct, the protein oligopeptide rate of recovery is 57%, controlled being made as of protein oligopeptide molecular weight (＞85%)≤5000Da do not evaluated the taste local flavor of protein oligopeptide; (2002) such as U.S. Azizah A.H use Alcalase proteolytic enzyme, and E/S=20g/kg extracts protein oligopeptide and the vacuum spray drying that obtains DH value 14.9% from tilapia meat; The effect that adopts Neutrase, Alcalase, Pepsin and Kojizyme protease hydrolyzed Atlantic Ocean cod and atlantic salmon processed side product steck has been compared in research such as Norway B.Liaset (2000), wherein adopt Alcalase proteolytic enzyme (E/S=30AU/kg), with Kojizyme proteolytic enzyme (E/S=5000LAPU/kg) two-step approach enzymolysis Atlantic salmon processed side product steck (material-water ratio 1: 2), the protein oligopeptide rate of recovery is up to 67%, controlled being made as of protein oligopeptide molecular weight (＞85%)≤1000Da, but the local flavor of protein oligopeptide is not discussed.At present, do not see that with Pacific Ocean wall pollack be the report that specified raw material prepares the enzymolysis process method of protein oligopeptide powder, utilize other fish to prepare the relevant enzymolysis processing method research of protein oligopeptide, still be confined to laboratory work, the enzymolysis process parameter is not optimized screening mostly, exists the low shortcoming of the protein oligopeptide powder rate of recovery.In addition, the relevant report of fish protein oligopeptide goods debitterize, deodorization technology seldom.
Summary of the invention:
The objective of the invention is to propose a kind of enzymolysis process method of utilizing Pacific Ocean wall pollack flesh of fish preparation protein oligopeptide powder, optimize and screened the enzymolysis process parameter, to improve the rate of recovery of protein oligopeptide powder, remove the bitter taste and the fishy smell of protein oligopeptide powder, obtain the protein oligopeptide powder of two kinds of different molecular weight ranges.
The present invention realizes by following technical scheme, its processing method is: Pacific Ocean wall pollack fish, decaptitate, go dirty, peeling, clean, rub the sterilization of heating of back input enzyme reactor through mincer, adding water and protease preparation again mixes, enzymolysis process parameter (the material-water ratio of optimizing, time, temperature, pH) enzymolysis under the condition, enzymolysis solution heats up and makes the proteolytic enzyme deactivation then, coarse filtration is separated spur, the centrifugal soluble proteins oligopeptides water that obtains respectively of the further three-phase of enzymolysis solution, insoluble protein precipitation and fish oil, soluble proteins oligopeptides water is purified through ultrafiltration, take off raw meat, obtain the protein oligopeptide powder after the spraying drying, insoluble protein precipitation drying obtains the byproduct protein powder.
Protease preparation is: the combination of Bacillus subtilus endo-protease and the circumscribed proteolytic enzyme of aspergillus tubigensis or genus bacillus compound protease; Protease preparation consumption E/S is: the Bacillus subtilus endo-protease is: 20AU/kg～80AU/kg, the circumscribed proteolytic enzyme of aspergillus tubigensis are: 8000LAPU/kg～40000LAPU/kg or genus bacillus compound protease are: 20 AU/kg～80 AU/kg.
Optimizing the enzymolysis process parameter is: material water blending ratio (w/v): 1.2: 1～1: 1.5; Hydrolysis temperature: 50 ℃～60 ℃; Enzymolysis time: 2hr～12hr; The initial pH value 6.6 of enzymolysis solution.
The raw meat that takes off of protein oligopeptide powder has adopted 8000rpm high speed centrifugation and the method that the ultrafiltration of 100kDa ultra-filtration membrane combines, and thoroughly holds back macromole fishy smell material.
Embodiment:
Be described in detail processing method of the present invention below by embodiment 1 and 2:
The present invention utilizes the enzymolysis process flow process of Pacific Ocean wall pollack flesh of fish preparation protein oligopeptide powder to be: select for use and decaptitate, go dirty, the cleaning Pacific Ocean wall pollack flesh of fish of peeling, be warming up to 95 ℃ after the rubbing, be incubated 10 ℃ of min sterilizations, mix with quantitative protease preparation and water again, in the time, temperature, enzymolysis in the enzyme reactor of controllable parameters such as pH, after enzyme digestion reaction finishes, enzymolysis solution is warming up to 10 ℃ of min of 90 ℃ of insulations and carries out deactivation, centrifugation, membrane sepn, each separates mutually spray-dried or oven dry, has both obtained target product: water-soluble protein oligopeptides powder and byproduct fish oil, fishbone powder, soluble protein powder.
It is protease preparation that the technology of the present invention adopts Bacillus subtilus endo-protease and the circumscribed proteolytic enzyme combination of aspergillus tubigensis or genus bacillus compound protease, has carried out the experiment of every batch of 200kg Pacific Ocean wall pollack flesh of fish preparation protein oligopeptide powder.The protein oligopeptide powder that obtains is light oyster white, no bitter taste, little fishy smell, and molecular weight (＞85%) can be respectively≤1000Da and 1000～5000Da, and the protein oligopeptide powder rate of recovery (raw material total protein content relatively) reaches 65%-79%.
Determining of protease preparation kind or protease preparation combination: the enzymolysis dynamics of proteolytic enzyme and combination thereof just plays topmost decisive action to the physicochemical property such as the bitter taste of protein digestion product, color and luster, molecular weight distribution and the protein oligopeptide powder rate of recovery.Therefore, it will be the key technical indexes with the bitter taste and the molecular weight distribution of enzymolysis product that proteolytic enzyme is selected, and carries out comprehensive evaluation.In the embodiment of the invention 1, proteolytic enzyme has adopted Bacillus subtilus endo-protease and the circumscribed proteolytic enzyme combination of aspergillus tubigensis; In the present embodiment 2, proteolytic enzyme has adopted the genus bacillus compound protease.After protein is hydrolyzed,, make enzymolysis solution produce bitter taste because the hydrophobic grouping and some the bitter taste amino acid that are hidden in long peptide chain of protein and the protein multidimensional structure are exposed.Zymin that is adopted among above-mentioned two embodiment or zymin combination all have the circumscribed characteristic of protein peptide chain, can decompose the hydrophobic grouping in the modified protein oligopeptides molecule, reach place to go bitter taste effect.Bitter taste removes with the hydrolysis of the Pacific Ocean wall pollack flesh of fish and finishes synchronously.
The E/S screening: should notice that higher E/S helps improving the protein oligopeptide powder rate of recovery, shortens enzymolysis time, and this effect will tend towards stability when E/S is too high near maximum, production cost then can increase because of the raising of proteolytic enzyme consumption on the other hand.E/S is respectively in the protease preparation combination of adopting in the present embodiment 1: the Bacillus subtilus endo-protease is that 50AU/kg, the circumscribed proteolytic enzyme of aspergillus tubigensis are that protease preparation E/S is among 30000LAPU/kg, the embodiment 2: the genus bacillus compound protease is 40AU/kg.
Optimize the enzymolysis process parameter: material-water ratio, hydrolysis temperature, enzymolysis time and initial pH value are the main technologic parameters that influences protease hydrolyzed efficient.Should notice that higher material-water ratio helps improving the extraction recovery of protein oligopeptide powder, but also can increase follow-uply concentrate, the energy consumption of drying process; Hydrolysis temperature is crossed to exceed to hang down and all can be reduced protease activities; Prolong enzymolysis time and can reduce the zymin consumption in right amount, and help improving the extraction recovery of protein oligopeptide, then reduced the turnover rate of utilization of equipment on the other hand; The initial pH of enzymolysis may influence the protease activities of enzymolysis process.Best material-water ratio, hydrolysis temperature, enzymolysis time and initial pH value make up, and the overall targets such as the protein oligopeptide powder rate of recovery, protein oligopeptide powder physical index, energy consumption of enzymolysis process method are optimized.In the present embodiment 1: material-water ratio (w/v) 1: 1,50 ℃ of hydrolysis temperatures, enzymolysis time: 5hr; The initial pH value of enzymolysis solution: 6.6.In the present embodiment 2: material-water ratio (w/v) 1: 1,55 ℃ of hydrolysis temperatures, enzymolysis time: 3hr; The initial pH value of enzymolysis solution: 6.6.
The centrifugal combination with ultra-filtration membrane taken off raw meat, method of purification: Pacific Ocean wall pollack flesh of fish fishy smell composition is mainly from fat, in the technological process of present embodiment 1 and embodiment 2, adopt the method for 8000rpm high speed centrifugation and 100kDa ultra-filtration membrane combining ultrafiltration, can hold back macromole fishy smell composition and allergen and heat source substance more up hill and dale.
This case process process is as follows,
Embodiment 1:
Select materials: the Pacific Ocean wall pollack flesh of fish (decaptitating, go dirty, peeling),
Protease preparation combination and E/S ratio are: Bacillus subtilus endo-protease 50AU/kg, the circumscribed protease 3 0000LAPU/kg of aspergillus tubigensis,
Optimize the enzymolysis process parameter: material-water ratio (w/v) 1: 1,50 ℃ of hydrolysis temperatures, enzymolysis time: 5hr; The initial pH value of enzymolysis solution: 6.6,
The centrifugal combination with ultra-filtration membrane taken off raw meat, method of purification: the method that adopts 8000rpm high speed centrifugation and 100kDa ultra-filtration membrane combining ultrafiltration.
Embodiment 2:
Select materials: the Pacific Ocean wall pollack flesh of fish (decaptitating, go dirty, peeling),
Protease preparation and E/S ratio are; Genus bacillus compound protease 40 AU/kg.
Optimize the enzymolysis process parameter: material-water ratio (w/v) 1: 1,55 ℃ of hydrolysis temperatures, enzymolysis time: 3hr; The initial pH value of enzymolysis solution: 6.6.
The centrifugal combination with ultra-filtration membrane taken off raw meat, method of purification: the method that adopts 8000rpm high speed centrifugation and 100kDa ultra-filtration membrane combining ultrafiltration.
Idiographic flow is:
That 200kg decaptitates, goes is dirty, the cleaning Pacific Ocean wall pollack fillet of peeling, after rubbing, drops into by mincer enzyme reactor, be warming up to 95 ℃ and be incubated 10min sterilization, embodiment 1 then: add water and protease preparation Bacillus subtilus endo-protease 50AU/kg, the circumscribed protease 3 0000LAPU/kg of aspergillus tubigensis.Material-water ratio (w/v) 1: 1,50 ℃ of hydrolysis temperatures, enzymolysis time 5hr, initial pH=6.6; Embodiment 2: add water and protease preparation genus bacillus compound protease 40 AU/kg.Material-water ratio (w/v) 1: 1,55 ℃ of hydrolysis temperatures, enzymolysis time 3hr, initial pH=6.6.After enzymolysis is finished, be warming up to 95 ℃ and be incubated 10min and carry out the proteolytic enzyme deactivation, enzymolysis solution is with after the bolting silk coarse filtration is separated fish bony spur (oven dry is pulverized and obtained fishbone powder), coarse filtration liquid centrifugal (8000rpm) under about 90 ℃ of temperature condition separates fish oil and water-insoluble albumen precipitation (oven dry is pulverized and obtained protein powder), the water-soluble protein oligopeptides water that obtains further after two times of ultrafiltration (ultra-filtration membrane molecular retention amount 100kDa and 1kDa/5kDa), the spray-dried protein oligopeptide powder product that promptly obtains of ultrafiltrated.
Protein oligopeptide powder product physicochemical property and main component that present embodiment 1,2 obtains see Table 1.
Table 1. protein oligopeptide powder product physicochemical property and main component
| The enzymolysis process method | The embodiment 1 50AU Bacillus subtilus endo-protease+circumscribed proteolytic enzyme of 30000LAPU aspergillus tubigensis, material-water ratio 1: 1, initial pH6.6,50 ℃ of temperature, enzymolysis 5hr, coarse filtration, ultrafiltration (100 kDa)+ultrafiltration (1kDa) | Embodiment 2 40AU genus bacillus compound proteases, material-water ratio 1: 1, initial pH6.6,55 ℃ of temperature, enzymolysis 3hr, coarse filtration, ultrafiltration (100kDa)+ultrafiltration (5kDa) |
| Oligopeptides molecular weight (＞85%), the DH value oligopeptide protein rate of recovery (%) color and luster bitter off-flavors are water-soluble | ≤ 1000Da, DH*=54% 72 light oyster whites do not have bitter taste, and little fishy smell is easily molten | 1000-5000Da, DH=23% 68 light oyster whites do not have bitter taste, and little fishy smell is easily molten |
Main component:
| Protein % fat % ash content % moisture % | ≥90 ≤0.1 ≤6.0 ≤4.0 | ≥90 ≤0.1 ≤6.0 ≤4.0 |
* protein degree: DH=% (free α-the total α of amino N/-amino N*100)
The present invention compares with existing correlation technique, has following characteristics:
1. Pacific Ocean wall pollack fish belongs to the ocean wild fish, is the white flesh of fish, and the foodsafety height can satisfy the requirement of the high-quality protein oligopeptide powder of preparation. Cost of material is moderate, and steady sources is easy to realize industrialization.
Screening and optimizing utilize the preparation of the Pacific Ocean wall pollack flesh of fish to extract the processes such as the enzyme preparation of protein oligopeptide, enzymolysis parameter, separating-purifying, protein oligopeptide powder rate of recovery height (relatively raw material total protein content) can reach 65%-79%. The technology maturity height such as the relevant enzyme reactor that requires, centrifugal, ultrafiltration, drying equipment, feasibility are strong.
By the enzyme preparation group and, enzymolysis parameter regulation and control, centrifugal and ultrafiltration membrance filter method, can obtain respectively two kinds of protein oligopeptide powder of molecular weight (＞85%)≤1000Da and 1000Da-5000Da, reach simultaneously the effect of the big molecule anaphylactogen of purifies and separates or heat source substance.
4. the protein oligopeptide powder does not have bitter taste, little fishy smell. Protein oligopeptide powder bitter taste removes with the flesh of fish hydrolysis of Pacific Ocean wall pollack and carries out synchronously, the usually absorption such as active carbon, covers up thorough, the easy economy of technological effect. Fishy smell elimination process has adopted centrifugal and physical method ultrafiltration membrance filter, more is conducive to guarantee natural sex and the security of product.
Claims (1)
1. one kind is utilized Pacific Ocean wall pollack to oppress the enzyme solution for preparing the protein oligopeptide powder, the method that it is characterized in that it is: the Pacific Ocean wall pollack flesh of fish of decaptitating, remove the peel, cleaning is after mincer rubs, drop into the enzyme reactor sterilization of heating, adding water and protease preparation again mixes, enzymolysis under the material-water ratio of optimizing, time, temperature, pH condition, enzymolysis solution heat up then with the proteolytic enzyme deactivation, coarse filtration is separated spur, centrifugal soluble proteins oligopeptides water, insoluble protein precipitation and the fish oil of obtaining respectively of the further three-phase of enzymolysis solution; Soluble proteins oligopeptides water is purified, is taken off raw meat through ultrafiltration, obtains the protein oligopeptide powder after the spraying drying; Insoluble protein precipitation drying obtains the byproduct protein powder; Described protease preparation is: the combination of Bacillus subtilus endo-protease and the circumscribed proteolytic enzyme of aspergillus tubigensis; Described protease preparation consumption E/S is: Bacillus subtilus endo-protease 20AU/kg～80AU/kg, the circumscribed proteinase 8 000LAPU/kg～40000LAPU/kg of aspergillus tubigensis; The material-water ratio of described optimization, time, temperature, pH are: material-water ratio 1.2: 1～1: 1.5, time 2hr～12hr, 50 ℃～60 ℃ of temperature, initial pH=6.6; The purification of described protein oligopeptide powder, taking off raw meat, is the method that adopts the 8000rpm high speed centrifugation to combine with the ultrafiltration of 100kDa ultra-filtration membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100241841A CN1291032C (en) | 2004-06-03 | 2004-06-03 | Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100241841A CN1291032C (en) | 2004-06-03 | 2004-06-03 | Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1603420A CN1603420A (en) | 2005-04-06 |
| CN1291032C true CN1291032C (en) | 2006-12-20 |
Family
ID=34663436
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100241841A Expired - Fee Related CN1291032C (en) | 2004-06-03 | 2004-06-03 | Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1291032C (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101326958B (en) * | 2008-07-22 | 2011-10-26 | 浙江大学 | Peptide feedstuff additive for regulating immunity function of sea water fish |
| CN101703155B (en) * | 2009-11-17 | 2012-01-11 | 中国水产科学研究院黄海水产研究所 | Preparation method and application of prawn non-specific immunity reinforcing agent oligopeptide |
| CN101731440B (en) * | 2010-02-10 | 2012-07-25 | 鞍山嘉鲜农业发展有限公司 | Composite all-fish protein powder and preparation method thereof |
| CN103494039B (en) * | 2013-10-10 | 2015-04-22 | 中国水产科学研究院黄海水产研究所 | Additive for reducing visceral fat deposition of weever and application thereof |
| CN103478434B (en) * | 2013-10-10 | 2015-01-07 | 中国水产科学研究院黄海水产研究所 | Novel composite protein source and its application in turbot feed |
| CN109678926A (en) * | 2019-02-25 | 2019-04-26 | 大连工业大学 | A kind of extracting method of solubility gadus albumen |
-
2004
- 2004-06-03 CN CNB2004100241841A patent/CN1291032C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1603420A (en) | 2005-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7422236B2 (en) | Method for producing clam active peptide | |
| Mirzapour-Kouhdasht et al. | Optimization of gelatin production from Barred mackerel by-products: Characterization and hydrolysis using native and commercial proteases | |
| CN101948899B (en) | Method for preparing antihypertensive peptides by using enzymatic degradation on mussel-digested protein | |
| CN106967169B (en) | Extraction method of fish collagen | |
| CN101897380A (en) | Preparation method of anchovy protein powder | |
| CN112481343B (en) | Method for producing collagen peptide by using cod skin raw material | |
| CN1883289A (en) | Method for preparing active polypeptide solution of fresh water fish protein | |
| CN111084346B (en) | Preparation process of sea cucumber tablet and sea cucumber tablet prepared by preparation process | |
| Anal et al. | Protein hydrolysates and bioactive peptides from seafood and crustacean waste: Their extraction, bioactive properties and industrial perspectives | |
| CN109722461B (en) | Deep sea fish collagen peptide and production method thereof | |
| CN107278208B (en) | Chitin, hydrolysate and production of at least one target product from insects by means of enzymatic hydrolysis | |
| CA3010453A1 (en) | A new method to improve enzyme hydrolysis and resultant protein flavor and bio-activity of fish offcuts | |
| CN1291032C (en) | Enzymolysis method for preparing protein oligopeptide powder using Pacific Ocean alaska pollack | |
| US4853231A (en) | Method for preparation of tastable matters consisting primarily of low molecular weight peptides | |
| CN1733925A (en) | Method with discarded protein Preparation Moriamin S | |
| CN113563459A (en) | Protein extraction process in production process of collagen peptide | |
| JP4227984B2 (en) | Composition for improving flavor and taste of food | |
| US7297512B2 (en) | Method for producing amino acid components by enzymatic hydrolysis of fish egg skin | |
| CN1234288C (en) | Process for preparation of protein hydrolysate from soy flour | |
| CN105002246A (en) | Fish polypeptide with uniform molecular weight distribution, and preparation method thereof | |
| CN115403668A (en) | Extraction process of bioactive collagen peptide of channel catfish swim bladder | |
| KR100612600B1 (en) | Process for preparing low molecular weight soybean peptide | |
| CA1325133C (en) | Method for preparation of tastable matters consisting mainly of low molecular weight peptides | |
| RU2226841C1 (en) | Protein food additive production method | |
| JPH03280835A (en) | Decomposition product of glair and food containing same decomposition product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |