CN104068204A - Preparation method of giant salamander polypeptides - Google Patents
Preparation method of giant salamander polypeptides Download PDFInfo
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- CN104068204A CN104068204A CN201410264105.8A CN201410264105A CN104068204A CN 104068204 A CN104068204 A CN 104068204A CN 201410264105 A CN201410264105 A CN 201410264105A CN 104068204 A CN104068204 A CN 104068204A
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Abstract
The invention discloses a preparation method of giant salamander polypeptides, belonging to the technical field of giant salamander deep processing as well as foods, health products and medicines. The preparation method comprises the following steps: preprocessing raw materials, boiling for softening giant salamander meat tissues, performing enzymolysis through compound protease, performing enzyme deactivation, and performing multistage filtering to obtain polypeptide product liquid of less than 20,000Da. According to the preparation method, a process is simple, the influence on an enzymolysis process and an enzymolysis product of epidermal cuticle of giant salamander can be effectively avoided by the boiling method, enzymolysis conditions are mild, giant salamander meat can be subjected to full enzymolysis, the enzymolysis process is easy to control, the enzymolysis product is safe and favorable for absorption by a human body, the added value of giant salamander is greatly improved, and the deep fine processing of giant salamander is facilitated.
Description
Technical field
The present invention relates to a kind of preparation method of giant salamander polypeptide, belong to giant salamander deep processing and food, health products and medical technical field.
Background technology
Giant salamander is the high animal of a kind of edibility, its fine and tender taste, delicious flavour, contain good protein, abundant amino acid and trace element, nutritive value is high, is described as " ginseng in water ", and giant salamander muscle protein is a kind of high-quality protein, essential amino acids content is high, proportion of composing is good, meets human body requirement pattern completely, and its nutritive value is much better than abalone, bird's nest, shark's fin and soft-shelled turtle.Giant salamander muscle protein is rich in 18 seed amino acids, wherein 6 kinds are taste amino acid and account for 42.77% of total amino acid content, 8 kinds of essential amino acids account for 40.72% of total amino acid content, and essential amino acid and nonessential amino acid ratio are 68.68 %, all meet the idealized model of FAO/WHO; The scoring of giant salamander essential amino acid is high, meets human body requirement pattern degree quite high, wherein contains the lysine easily lacking in abundant China's staple food, can be reasonably combined edible with staple food, and to play protein complementary action.
The eating method of artificially-cultured giant salamander is mainly the mode that adopts conventional cooking, is limited to the digestion desorption system of human body, and the nutritive value of giant salamander can not be fully utilized.Along with the development of biotechnology, occur giant salamander to carry out deep process.But because the crust of giant salamander has black cuticula, processing is impacted, meat entirety breaking method is carried out in general employing, but cuticular existence makes enzymolysis process not thorough, simultaneously because the too small meeting of cuticula impacts enzymolysis process and product.
Summary of the invention
Goal of the invention of the present invention is: for the problem of above-mentioned existence, the preparation method of the convenient and efficient giant salamander polypeptide of a kind of technique is provided, can avoid the impact of paying no attention to that giant salamander cuticula causes polypeptide yield and quality in traditional preparation method, utilize rational enzyme to improve enzymolysis quality simultaneously, be beneficial to the absorption of human body.
The technical solution used in the present invention is as follows:
A preparation method for giant salamander polypeptide, is characterized in that: comprise the steps:
(a) pretreatment: giant salamander is cleaned and stripping and slicing;
(b) infusion: add water to gained giant salamander cube meat, infusion is to giant salamander cube meat organization softening;
(c) enzymolysis: after feed liquid after infusion is cooling, moisturizing regulates solid-liquid ratio, then adds compound protease to carry out enzymolysis, and described compound protease comprises neutral proteinase and papain;
(d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the enzyme that goes out;
(e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, the pure polypeptide products of system.
Carrying out before above-mentioned processing, it is emptied the intestine work giant salamander a couple of days of stopping eating, then slaughter gall-bladder, tissue is for the preparation of giant salamander polypeptide entirely.
Further, in described step (a), giant salamander is cut into 0.5-2cm
3piece.
Further, in step (b), giant salamander cube meat is in the infusion that adds water of 1:1-2 ratio, and the preferred infusion temperature of described infusion process is 111 DEG C-123 DEG C, and the time is 20min-30min.
Further, in described step (c), described neutral proteinase is 50,000 U/g, and consumption is the 0.5%-1.5% of giant salamander cube meat quality, and papain is 100,000 U/g, and consumption is the 0.6%-2.0% of giant salamander cube meat quality.
Further, described neutral protein enzyme dosage is 1.2% of giant salamander cube meat quality, and papain consumption is 1.6% of giant salamander cube meat quality.
Further, described compound protease also comprises flavor protease, and described flavor protease is made up of with mass ratio 3-5:1 aminopeptidase and carboxypeptidase, is preferably 4:1, and flavor protease consumption is the 0.3-1.0% of giant salamander cube meat quality, and preferable amount is 0.8%.
Further, described step (c) moisturizing regulates in solid-liquid ratio, and regulating the ratio of raw material giant salamander cube meat and water is 1:2-6.
Further, the mode that in described step, the employing of (c) is stirred is carried out enzymolysis, and mixing speed is 400r/min, and hydrolysis temperature is that 45 DEG C-55 DEG C, enzymolysis time are 4-8 hour, and enzymolysis PH is 6.5-7.5.
Further, in described step (d), enzyme-deactivating condition is deactivation 15min at 90 DEG C-100 DEG C.
Further, in described step in (e), adopt three grades of step apertures to filter, 30-50 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is that the milipore filter of 20000Da carries out ultrafiltration.
The giant salamander polypeptide that adopts described method to prepare.
Adopt giant salamander polypeptide prepared by the described method application in drink, health products and medicine.
In sum, owing to having adopted technique scheme, the invention has the beneficial effects as follows: the preparation method of described giant salamander polypeptide, technique is simple, adopts infusion method can effectively avoid giant salamander horny layer of epidermis on enzymolysis process and enzymolysis product impact, enzymatic hydrolysis condition gentleness, can carry out sufficient enzymolysis to Megalobatrachus japonicus daoidianuas(Blanchard), enzymolysis process is easy to control, and enzymolysis product safety and be beneficial to absorption of human body, has improved the added value of giant salamander greatly.
Detailed description of the invention
Disclosed all features in this description, or step in disclosed all methods or process, except mutually exclusive feature and/or step, all can combine by any way.
Disclosed arbitrary feature in this description, unless narration especially all can be replaced by other equivalences or the alternative features with similar object.,, unless narration especially, each feature is an example in a series of equivalences or similar characteristics.
It is emptied the intestine work giant salamander a couple of days of stopping eating, then slaughter gall-bladder, tissue is for the preparation of giant salamander polypeptide entirely.
embodiment 1
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: giant salamander is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:1 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:2 taking the quality moisturizing of raw material giant salamander cube meat, add again the compound protease being formed by neutral proteinase and papain to carry out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is the 0.5%(0.25kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 0.6%(0.3kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 45 DEG C, enzymolysis time is 4 hours, enzymolysis is to carry out for 6.5 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 30 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
embodiment 2
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: giant salamander is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:1.5 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:4 taking the quality moisturizing of raw material giant salamander cube meat, add again the compound protease being formed by neutral proteinase and papain to carry out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is 1.2 %(0.6kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 1.6%(0.8kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 50 DEG C, enzymolysis time is 6 hours, enzymolysis is to carry out for 7.0 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 40 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
embodiment 3
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: giant salamander is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:2 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:6 taking the quality moisturizing of raw material giant salamander cube meat, add again the compound protease being formed by neutral proteinase and papain to carry out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is the 1.5%(0.75kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 2.0%(1kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 55 DEG C, enzymolysis time is 8 hours, enzymolysis is to carry out for 7.5 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 50 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
embodiment 4
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: Megalobatrachus japonicus daoidianuas(Blanchard) is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:1 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:2 taking the quality moisturizing of raw material giant salamander cube meat, add again by neutral proteinase, the compound protease of papain and flavor protease composition carries out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is the 0.5%(0.25kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 0.6%(0.3kg of giant salamander cube meat quality), flavor protease is made up of with mass ratio 3:1 aminopeptidase and carboxypeptidase, aminopeptidase and carboxypeptidase are 1.5 ten thousand U/g, flavor protease consumption is the 0.3%(0.15kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 45 DEG C, enzymolysis time is 4 hours, enzymolysis is to carry out for 6.5 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 30 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
embodiment 5
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: giant salamander is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:1.5 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:4 taking the quality moisturizing of raw material giant salamander cube meat, add again by neutral proteinase, the compound protease of papain and flavor protease composition carries out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is 1.2 %(0.6kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 1.6%(0.8kg of giant salamander cube meat quality), flavor protease is made up of with mass ratio 5:1 aminopeptidase and carboxypeptidase, aminopeptidase and carboxypeptidase are 1.5 ten thousand U/g, flavor protease consumption is the 1.0%(0.5kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 50 DEG C, enzymolysis time is 6 hours, enzymolysis is to carry out for 7.0 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 40 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
embodiment 6
The preparation method of the present embodiment giant salamander polypeptide, comprises the steps: (a) pretreatment: giant salamander is cleaned and is cut into 0.5-2cm
3bulk, get 50kg processing, (b) infusion: add water with the amount of 1:2 to gained giant salamander cube meat, infusion 20min-30min at 111 DEG C-123 DEG C, to giant salamander cube meat organization softening, (c) enzymolysis: feed liquid after infusion with pressure naturally cooling after, regulate material-water ratio as 1:6 taking the quality moisturizing of raw material giant salamander cube meat, add again by neutral proteinase, the compound protease of papain and flavor protease composition carries out enzymolysis, described neutral proteinase is 50,000 U/g, consumption is the 1.5%(0.75kg of giant salamander cube meat quality), papain is 100,000 U/g, consumption is the 2.0%(1kg of giant salamander cube meat quality), flavor protease is made up of with mass ratio 4:1 aminopeptidase and carboxypeptidase, aminopeptidase and carboxypeptidase are 1.5 ten thousand U/g, flavor protease consumption is the 0.8%(0.4kg of giant salamander cube meat quality), enzymolysis adopts the mode stirring to carry out, mixing speed is 400r/min, hydrolysis temperature is 55 DEG C, enzymolysis time is 8 hours, enzymolysis is to carry out for 7.5 times at PH, (d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the deactivation 15min enzyme that goes out at 90 DEG C-100 DEG C, (e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, first adopt three grades of step apertures to filter, 50 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is the pure polypeptide products that the milipore filter of 20000Da carries out ultrafiltration system.
The preparation method of giant salamander polypeptide of the present invention, technique is simple, adopt infusion method can effectively avoid giant salamander horny layer of epidermis on enzymolysis process and enzymolysis product impact, enzymatic hydrolysis condition gentleness, can carry out enzymolysis fully efficiently to Megalobatrachus japonicus daoidianuas(Blanchard), enzymolysis process is easy to control, and enzymolysis product safety and be beneficial to absorption of human body, has improved the added value of giant salamander greatly; Flavor protease cuts off the peptide bond of polypeptide inside by endo protease, form short-chain peptide, some of them contain hydrophobic amino acid thereby become bitter peptide, excision enzyme cuts off and discharges an amino acid from the end of polypeptide chain each time, thereby bitter peptide is thoroughly degraded to amino acid, the bitter taste that the introducing of flavor protease can be controlled polypeptide liquid, improves the quality of products.
The present invention is not limited to aforesaid detailed description of the invention.The present invention expands to any new feature or any new combination disclosing in this manual, and the arbitrary new method disclosing or step or any new combination of process.
Claims (10)
1. a preparation method for giant salamander polypeptide, is characterized in that: comprise the steps:
(a) pretreatment: giant salamander is cleaned and stripping and slicing;
(b) infusion: add water to gained giant salamander cube meat, infusion is to giant salamander cube meat organization softening;
(c) enzymolysis: after feed liquid after infusion is cooling, moisturizing regulates solid-liquid ratio, then adds compound protease to carry out enzymolysis, described compound protease comprises neutral proteinase and papain composition;
(d) enzyme that goes out: after enzymolysis completes, enzymolysis liquid is boiled to the enzyme that goes out;
(e) filter: the feed liquid of going out after enzyme is carried out to multistage filtering, the pure polypeptide products of system.
2. the preparation method of giant salamander polypeptide as claimed in claim 1, is characterized in that: in described step (a), giant salamander is cut into 0.5-2cm
3piece.
3. the preparation method of giant salamander polypeptide as claimed in claim 1 or 2, it is characterized in that: in described step (c), described neutral proteinase is 50,000 U/g, and consumption is the 0.5-1.5% of giant salamander cube meat quality, papain is 100,000 U/g, and consumption is the 0.6-2.0% of giant salamander cube meat quality.
4. the preparation method of giant salamander polypeptide as claimed in claim 3, is characterized in that: described neutral protein enzyme dosage is 1.2% of giant salamander cube meat quality, and papain consumption is 1.6% of giant salamander cube meat quality.
5. the preparation method of the giant salamander polypeptide as described in claim 1 to 4 any one, it is characterized in that: described compound protease also comprises flavor protease, described flavor protease is made up of with mass ratio 3-5:1 aminopeptidase and carboxypeptidase, and flavor protease consumption is the 0.3-1.0% of giant salamander cube meat quality.
6. the preparation method of the giant salamander polypeptide as described in claim 1 to 5 any one, is characterized in that: described step (c) moisturizing regulates in solid-liquid ratio, and regulating the ratio of giant salamander cube meat and water is 1:2-6.
7. the preparation method of the giant salamander polypeptide as described in claim 1 to 6 any one, it is characterized in that: the mode that in described step, the employing of (c) is stirred is carried out enzymolysis, mixing speed is 400r/min, and hydrolysis temperature is that 45 DEG C-55 DEG C, enzymolysis time are 4-8 hour, and enzymolysis PH is 6.5-7.5.
8. the preparation method of the giant salamander polypeptide as described in claim 1 to 7 any one, it is characterized in that: in described step in (e), adopt three grades of step apertures to filter, 30-50 μ m, 5 μ m, the 0.45 μ m of being followed successively by different levels, finally adopting interception is that the milipore filter of 20000Da carries out ultrafiltration.
9. the giant salamander polypeptide that as described in claim 1 to 8 any one prepared by method.
10. the application of giant salamander polypeptide in drink, health products and medicine as claimed in claim 9.
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| CN105018555A (en) * | 2015-08-06 | 2015-11-04 | 重庆馗丰食品有限公司 | Preparation method of giant salamander skin collagen peptide |
| CN107259582A (en) * | 2017-07-06 | 2017-10-20 | 长沙爱扬医药科技有限公司 | Giant salamander protein peptide powder product and preparation method thereof |
| CN107326056A (en) * | 2017-07-21 | 2017-11-07 | 重庆贡龙大鲵生物科技有限公司 | The preparation method of giant salamander polypeptide powder |
| CN107446976A (en) * | 2017-07-21 | 2017-12-08 | 重庆贡龙大鲵生物科技有限公司 | The preparation method of giant salamander polypeptide powder |
| CN109385457A (en) * | 2018-11-07 | 2019-02-26 | 四川大学 | A kind of preparation method of the giant salamander Mei Lade peptide with antioxidant activity |
| CN111172226A (en) * | 2020-03-09 | 2020-05-19 | 中健智诊(重庆)生物研究院 | Collagen peptide for intestinal absorption and extraction method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105018555A (en) * | 2015-08-06 | 2015-11-04 | 重庆馗丰食品有限公司 | Preparation method of giant salamander skin collagen peptide |
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| CN109385457A (en) * | 2018-11-07 | 2019-02-26 | 四川大学 | A kind of preparation method of the giant salamander Mei Lade peptide with antioxidant activity |
| CN109385457B (en) * | 2018-11-07 | 2022-02-11 | 四川大学 | Preparation method of giant salamander Maillard peptide with antioxidant activity |
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Application publication date: 20141001 |