CN103981246A - Method for extracting bitter peptides from fish protein enzymolysis solution - Google Patents
Method for extracting bitter peptides from fish protein enzymolysis solution Download PDFInfo
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- CN103981246A CN103981246A CN201410234947.9A CN201410234947A CN103981246A CN 103981246 A CN103981246 A CN 103981246A CN 201410234947 A CN201410234947 A CN 201410234947A CN 103981246 A CN103981246 A CN 103981246A
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Abstract
The invention relates to a method for extracting bitter peptides from a fish protein enzymolysis solution, belonging to the technical field of marine organism processing. The method comprises the steps of raw material pretreatment, enzymolysis, centrifugation and chromatography, and specifically comprises the following steps: cleaning fish or fish offal, mashing, adding water, uniformly mixing, regulating the pH value, adding a proteinase, carrying out enzymolysis under certain conditions, centrifugating with a three-leg centrifugal machine to remove the residues and grease so as to obtain an enzymolysis supernate, passing the enzymolysis supernate through a macroporous resin chromatographic column, passing distilled water through the macroporous resin chromatographic column until no colored liquid flows out, eluting the macroporous resin chromatographic column with 20-80% ethanol, collecting the eluate, and volatilizing to remove the ethanol, thereby obtaining the bitter peptides. The method can simultaneously extract the bitter peptides and prepare the low-bitterness protein enzymolysis solution; and in the prepared bitter peptides, the peptide content is higher than 95%, the free amino acid content is lower than 5%, and the content of the peptides with the molecular weight of lower than 2000Da is about 80%. The method is simple in process and easy to operate, has the advantages of low cost and high safety and reliability, and can easily implement industrialization.
Description
Technical field
The present invention relates to a kind of extracting method of bitter peptides, be specifically related to a kind of method of fish protein enzymolysis solution extraction bitter peptides, belong to marine organisms processing technique field.
Background technology
Fish protein enzymolysis solution contains abundant polypeptide, little peptide, amino acid, the multiple small molecules nutritive ingredients such as trace element, there is bioavailability advantages of higher, also have anti-oxidant, hypotensive, strengthen immune isoreactivity, it can be widely used as food flavouring, puffed food and convenient, the base-material of protective foods, also can be used as patient's foodstuff, its product can be with powdery, the variform such as liquid and pasty state exists, be convenient to according to different physiological status crowds, crowd's nutritional character and market requirement situation under different work condition, fully mix with other food material, again be processed into colourful oceanographic engineering food, there is good development prospect, market potential is huge.
Because enzymolysis meeting produces low molecule bitter peptides, bitter peptides is rich in hydrophobic amino acid, makes enzymolysis solution bitter taste outstanding, has limited its application in foodstuffs industry, becomes the bottleneck that fish enzymolysis protein develops.Some hydrophobic amino acids in bitter peptides as the whiles such as leucine, Isoleucine, α-amino-isovaleric acid, phenylalanine be again indispensable amino acid, therefore there is higher nutritive value, bitter peptides is also some small-molecular peptides, there is equally the good mechanism of absorption of peptide class and physics and chemistry processing characteristics, also there is some biological activity.The research of enzymolysis solution bitter taste at present concentrates on debittering method mostly, there is the embedding encapsulation method of cyclodextrin, gelatin, treated starch and pectin etc., the biological debittering france of microorganism fermentation, generates hydrophobic amino acid enrichment by transpeptidation the plastein reaction of water-fast hydrophobic peptide etc.Also there is patent to disclose and utilize macroporous adsorbent resin preparation to take off the technology of raw meat protein peptide or oligopeptide.Patent " is utilized macroporous adsorbent resin to produce the method (CN201010028931) of no-fishy smell scale collagen peptide " and is disclosed a kind of method of producing no-fishy smell scale collagen peptide, to adsorb through macroporous adsorptive resins through the collagen peptide hydrolyzed solution of activated carbon decolorizing and filtration clarification, first through ethanolic soln wash-out fishy smell material, by ammoniacal liquor wash-out collagen peptide, through membrane sepn, concentrated, obtain no-fishy smell scale collagen peptide through lyophilize again, ethanol eluate is dried and obtains fishy smell material through ethanol recovery, spraying.Here do not indicate specifically what material of fishy smell material, use ammoniacal liquor wash-out through membrane sepn, concentrated simultaneously, then obtain no-fishy smell scale collagen peptide through lyophilize, complex process, cost is high; Patent " a kind of phytic acid and oligopeptide (CN201010520721) prepared taking defatted rice bran as raw material coproduction " discloses a kind of method that defatted rice bran is prepared phytic acid and oligopeptide, low-concentration hcl rice dipping chaff is extracted to the precipitation after phytic acid, adopt Sumizyme MP and compound protease fractional hydrolysis, enzymolysis solution is through ultrafiltration, macroporous adsorbent resin separation and purification, after lyophilize, obtain rice bran oligopeptide.In technique, enzymolysis solution is first adjusted pH3.0-5.0, by absorption with macroporous adsorbent resin, then uses 75% ethanol elution, boils off ethanol lyophilize and obtains rice bran oligopeptide.Molecular weight composition and the relevant physicochemical characteristics of oligopeptide are not described here.The method of bitter peptides is prepared in extraction provided by the invention, by fish protein enzymolysis solution with the certain flow rate macroporous adsorptive resins of flowing through, the low raw meat bitter taste of effluent liquid, then extract and prepare bitter peptides with ethanol elution adsorption resin column, the physicochemical characteristics of bitter peptides is clear and definite.Technological process is simple, does not need adjust pH, does not need ammoniacal liquor wash-out, not only can obtain low molecule bitter peptides, can also remove the bitter taste of enzymolysis solution, for the exploitation of fish protein enzymolysis solution and bitter peptides provide basis.
Summary of the invention
The object of this invention is to provide a kind of method that fish protein enzymolysis solution extracts bitter peptides, the method technique is simple, with low cost, adopts the method can obtain low molecule bitter peptides, and can remove the taste of enzymolysis solution.
In order to address the above problem, the technical solution adopted in the present invention is:
Fish protein enzymolysis solution extracts a method for bitter peptides, specifically comprises the steps:
(1) raw materials pretreatment: blend after fish or fish scrap are cleaned, add water and mix, and to regulate pH value be 5.5 ~ 8.0;
(2) enzymolysis: add proteolytic enzyme in the raw material mixing, and carry out enzymolysis under certain condition, the cooling enzymolysis solution that obtains;
(3) centrifugal: by centrifugal link-suspended basket centrifuge for enzymolysis solution (being furnished with filter bag) residue and the grease of removing, to obtain supernatant liquor;
(4) chromatography: enzymolysis supernatant liquor is with the certain flow rate macroporous resin chromatography column of flowing through, flow through macroporous resin chromatography column with distilled water again until have or not color liquid to flow out, then use the ethanol elution macroporous resin chromatography column of 20%-80%, collect elutriant, volatilization goes ethanol to obtain bitter peptides.
The mixing that adds water that step (1) is described, the ratio of the water adding and fish or fish scrap is: 1:2 ~ 1:5.
The described enzymolysis of step (2) refers to that adding massfraction is the proteolytic enzyme of mixing raw material 0.3% ~ 0.65%, at 45 DEG C ~ 55 DEG C temperature, and enzymolysis 1～6 hour, enzymolysis finishes, and is warming up to 90 DEG C～95 DEG C enzymes 15 minutes of going out, and cooling.
The described proteolytic enzyme of step (2) is the one, two or three in neutral protease, food flavor enzyme, bromeline, papoid and trypsinase.
The described link-suspended basket centrifuge of step (3) is food-processing or pharmaceutical purpose whizzer,
The described macroporous resin of step (4) is food grade macroporous adsorbent resin.
Effective effect of the present invention is:
(1) a kind of fish protein enzymolysis solution provided by the invention extracts the method for bitter peptides, and technological process is simple, processing ease, and cost is low;
(2) a kind of fish protein enzymolysis solution provided by the invention extracts the method for bitter peptides, and in the bitter peptides of preparation, the content of peptide is more than 95%, and free aminoacid content is below 5%, and the peptide below 2000Da accounts for 80%;
(3) a kind of fish protein enzymolysis solution provided by the invention extracts the method for bitter peptides, has prepared the protein enzymatic hydrolyzate of low raw meat bitter taste in extracting bitter peptides, for the exploitation of protein enzymatic hydrolyzate provide basis;
(4) a kind of fish protein enzymolysis solution provided by the invention extracts the method for bitter peptides, the macroporous resin using, and regeneration is convenient, can Reusability, safe and reliable, be easy to realize industrialization and produce.
Embodiment
Below by embodiment, the present invention is described in further details, these embodiment are only used for illustrating the present invention, do not limit the scope of the invention.
embodiment 1
Get leftovers of tilapia 2kg, clean and go after internal organ, after mincer rubs, put into reactor, add the water that rubs 3 times of Tilapia weight, adjust pH is 7.0, adds the neutral protease of raw material weight 0.25% and 0.3% food flavor enzyme, enzymolysis 4h simultaneously, hydrolysis finishes, be warming up to 95 DEG C of enzymes 15 minutes of going out, obtain enzymolysis solution after cooling, centrifugal through the link-suspended basket centrifuge (joining 600 object filter bags) of rotating speed 3000r/min, remove residue and grease, collect filtered liquid A; By 6BV volume filtered liquid A taking protein concn 7.2mg/mL, the flow velocity 5BV/h AB-8 macroporous adsorption resin chromatography post of resin bed volume as BV of flowing through; With 5BV distilled water, the filtrate A not being adsorbed in post is rinsed out again; Then with 40% ethanolic soln, the 3BV/h flow velocity of 12BV flow through chromatography column wash-out extract, collect elutriant, volatilization ethanol, obtain bitter peptides 16.6 g, yield 0.83%.In bitter peptides, peptide content is that small peptide content below 97.8%, 2000Da accounts for 75.2%, and the 8 seed amino acid content such as the hydrophobic amino proline acid relevant with bitter taste, phenylalanine, leucine reach 40.8%.
embodiment 2
Get Tilapia 2kg, after mincer rubs, put into reactor, add the water that rubs 2 times of Tilapia weight, adjust pH is 6.8, add the neutral protease of raw material weight 0.35% and 0.2% food flavor enzyme simultaneously, enzymolysis 3.5h, hydrolysis finishes, and is warming up to 95 DEG C of enzymes 15 minutes of going out, after cooling, obtain enzymolysis solution, link-suspended basket centrifuge (joining 800 object filter bags) through rotating speed 3000r/min is centrifugal, removes residue and grease, collects filtered liquid A; By 10BV volume filtered liquid A taking protein concn 7.2mg/mL, the flow velocity 3BV/h DA201-C macroporous adsorption resin chromatography post of resin bed volume as BV of flowing through; With 5BV distilled water, the filtrate A not being adsorbed in post is rinsed out again; Then with 60% ethanolic soln, the 3BV/h flow velocity of 12BV flow through chromatography column wash-out extract, collect elutriant, volatilization ethanol, obtain bitter peptides 21.0g, yield 1.05%.In bitter peptides, peptide content is that small peptide content below 95.6%, 2000Da accounts for 81.3%, and the 8 seed amino acid content such as the hydrophobic amino proline acid relevant with bitter taste, phenylalanine, leucine reach 36.2%.
Claims (6)
1. fish protein enzymolysis solution extracts a method for bitter peptides, it is characterized in that: specifically comprise the steps:
(1) raw materials pretreatment: blend after fish or fish scrap are cleaned, add water and mix, and to regulate pH value be 5.5 ~ 8.0;
(2) enzymolysis: add proteolytic enzyme in the raw material mixing, and carry out enzymolysis under certain condition, the cooling enzymolysis solution that obtains;
(3) centrifugal: by centrifugal residue and the grease of removing of link-suspended basket centrifuge of being furnished with filter bag for enzymolysis solution, to obtain supernatant liquor;
(4) chromatography: enzymolysis supernatant liquor is with the certain flow rate macroporous resin chromatography column of flowing through, flow through macroporous resin chromatography column until flow out without color liquid with distilled water again, then use the ethanol elution macroporous resin chromatography column of 20%-80%, collect elutriant, volatilization goes ethanol to obtain bitter peptides.
2. a kind of fish protein enzymolysis solution according to claim 1 extracts the method for bitter peptides, it is characterized in that: the mixing that adds water that step (1) is described, the ratio of the water adding and fish or fish scrap is: 1:2 ~ 1:5.
3. a kind of fish protein enzymolysis solution according to claim 1 extracts the method for bitter peptides, it is characterized in that: the described enzymolysis of step (2) refers to that adding massfraction is the proteolytic enzyme of mixing raw material 0.3% ~ 0.65%, at 45 DEG C ~ 55 DEG C temperature, enzymolysis 1～6 hour, enzymolysis finishes, be warming up to 90 DEG C～95 DEG C enzymes 15 minutes of going out, and cooling.
4. a kind of fish protein enzymolysis solution according to claim 1 extracts the method for bitter peptides, it is characterized in that: the described proteolytic enzyme of step (2) is the one, two or three in neutral protease, food flavor enzyme, bromeline, papoid and trypsinase.
5. a kind of fish protein enzymolysis solution according to claim 1 extracts the method for bitter peptides, it is characterized in that: the described link-suspended basket centrifuge of step (3) is food-processing or pharmaceutical purpose whizzer.
6. a kind of fish protein enzymolysis solution according to claim 1 extracts the method for bitter peptides, it is characterized in that: the described macroporous resin of step (4) is food grade macroporous adsorbent resin.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105803023A (en) * | 2016-05-10 | 2016-07-27 | 中国科学院南海海洋研究所 | Method for preparing aquatic livestock small molecule peptide without fishy smell or bitter taste |
| CN106279350A (en) * | 2016-08-11 | 2017-01-04 | 西南大学 | A kind of preparation method and application of Mucor type Semen Sojae Preparatum bitter peptides |
| CN106923129A (en) * | 2015-12-29 | 2017-07-07 | 中粮集团有限公司 | It is a kind of to cover soya-bean polypeptides solid beverage of bad flavor and preparation method thereof |
| CN107173778A (en) * | 2017-07-10 | 2017-09-19 | 浙江顶味食品有限公司 | A kind of meat albumen enzymolysis product for removing bitter taste, meat flavor and preparation method thereof |
| CN108030101A (en) * | 2017-11-08 | 2018-05-15 | 金华市飞凌生物科技有限公司 | The method of abstraction function small peptide from low value ocean fish |
| CN114480542A (en) * | 2022-01-27 | 2022-05-13 | 河南工业大学 | Method for extracting bitter peptides from wheat gluten protein enzymolysis products |
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| CN101736066A (en) * | 2010-01-08 | 2010-06-16 | 武汉普赛特膜技术循环利用有限公司 | Method for producing no-fishy smell scale collagen peptide by utilizing macroporous absorbent resin |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106923129A (en) * | 2015-12-29 | 2017-07-07 | 中粮集团有限公司 | It is a kind of to cover soya-bean polypeptides solid beverage of bad flavor and preparation method thereof |
| CN105803023A (en) * | 2016-05-10 | 2016-07-27 | 中国科学院南海海洋研究所 | Method for preparing aquatic livestock small molecule peptide without fishy smell or bitter taste |
| CN106279350A (en) * | 2016-08-11 | 2017-01-04 | 西南大学 | A kind of preparation method and application of Mucor type Semen Sojae Preparatum bitter peptides |
| CN106279350B (en) * | 2016-08-11 | 2019-08-16 | 西南大学 | A kind of preparation method and application of Mucor type fermented soya bean bitter peptides |
| CN107173778A (en) * | 2017-07-10 | 2017-09-19 | 浙江顶味食品有限公司 | A kind of meat albumen enzymolysis product for removing bitter taste, meat flavor and preparation method thereof |
| CN108030101A (en) * | 2017-11-08 | 2018-05-15 | 金华市飞凌生物科技有限公司 | The method of abstraction function small peptide from low value ocean fish |
| CN114480542A (en) * | 2022-01-27 | 2022-05-13 | 河南工业大学 | Method for extracting bitter peptides from wheat gluten protein enzymolysis products |
| CN114480542B (en) * | 2022-01-27 | 2024-05-28 | 河南工业大学 | Method for extracting bitter peptide from wheat gluten enzymolysis product |
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Application publication date: 20140813 |