CN108753897B - A kind of preparation method of sea cucumber internal organ polypeptide - Google Patents
A kind of preparation method of sea cucumber internal organ polypeptide Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method of sea cucumber internal organ polypeptide, step includes: that sea cucumber internal organ frozen goods are successively thawed, are digested, are centrifuged, are concentrated, drying steps, and complex enzyme is used in the enzymolysis step, and the complex enzyme is lipase and proteinase combination.The complex enzyme is the combination of Collagenase, papain and lipase.The utility model has the advantages that the present invention can significantly improve the flavor of sea cucumber internal organ polypeptide product, including smell and mouthfeel；The color of sea cucumber internal organ polypeptide product can be effectively improved；Yield can be improved, reduce the cost of product.
Description
Technical field
The present invention relates to food processing fields, and in particular to a kind of preparation method of sea cucumber internal organ polypeptide.
Background technique
Sea cucumber is precious precious marine product, hitherto it is found that containing in sea cucumber body there are many physiologically active ingredient, is had high
Food therapy value.Sea cucumber internal organ include alimentary canal, gonad, respiratory trees, occupy Vickers pipe, water lung, Polian vesicle, water ring pipe, tentacle
And sieve plate etc..The alimentary canal of sea cucumber, that is, sea cucumber intestinal wall and sea cucumber sexual gland are the important components of sea cucumber internal organ, and sea
Participate in the main waste of work.The most important ingredient of sea cucumber internal organ is protein, while raw rich in polysaccharide, saponin(e, flavonoids etc.
Active substances.Sea cucumber sexual gland is otherwise known as sea cucumber, and Japan and other countries and area are as invigorant, and based on intestines and internal organs of the body
Other sea cucumber internal organ be difficult to receive due to smell mouthfeel, processing and utilization is extremely limited.
Research shows that sea cucumber polypeptide has the bioactivity such as anti-oxidant, blood pressure lowering, have in medicine with field of health care food
Very big utility value.Sea cucumber internal organ flavor itself is poor, results in disorganization by enzymatic hydrolysis, the release of bad flavor adds
It is acute.The existing universal smell of sea cucumber polypeptide product produced using sea cucumber internal organ as raw material is compared with raw meat, irritant smell, mouth on the market
The peppery mouth of bitter and puckery flavor, have not pleasing mouth-feel, color be in crineous more, whole sensory-acceptance is poor.Sea cucumber polypeptide product is not
Good flavor mechanism is still not clear, just at present research known to may with the exposure of polypeptide hydrophobic grouping, system complicated component, saponin(e with it is more
The reasons such as sugared isoreactivity content of material is abundant are related, while having the fishy smell of marine products waste.Meanwhile existing sea cucumber polypeptide produces
Product are mostly dark brown, and color is obscure, influence its use.
For the flavor for improving sea cucumber internal organ polypeptide, most efficient method is that raw material is thoroughly cleaned or floated at present
It scalds, however will cause the heavy losses of yield being greatly reduced with active material in this way；The methods of physical absorption, ultrafiltration deodorant are de-
Color effect is limited and equally will cause the reduction of yield.Sea cucumber internal organ polypeptide product is typically only capable to addition essence, flavor improvement
Its bad flavor is covered up in agent, and covers up that effect is undesirable, and color is also difficult to improve, and constrains it in functional food and health care
Application in food.The acceptable degree of sense organ and yield for improving sea cucumber polypeptide simultaneously are that current sea cucumber polypeptide is prepared in industry urgently
It solves the problems, such as.
Summary of the invention
For the above-mentioned prior art, the present invention provides a kind of sea cucumber internal organ polypeptide production methods.The sea of this method preparation
Ginseng polypeptide smell, taste, mouthfeel, color significantly improve compared to existing product, effectively increase sea cucumber internal organ polypeptide system
The acceptable degree of the audient of product, while yield is higher.
The technical scheme to solve the above technical problems is that
A kind of preparation method of sea cucumber internal organ polypeptide, step include: sea cucumber internal organ frozen goods are successively thawed, enzyme
Solution, is concentrated, drying steps centrifugation, and complex enzyme is used in the enzymolysis step, and the complex enzyme is Collagenase, pawpaw egg
The combination of white enzyme and lipase；The weight ratio of the Collagenase, papain and lipase is 1:1:(1-2).
Preferably, the enzymatic hydrolysis condition are as follows: Collagenase, papain and lipase stir under conditions of pH7,
After digesting 3.5h under the conditions of 40 DEG C -50 DEG C, high temperature enzyme deactivation.
Preferably, the enzymatic hydrolysis condition are as follows: Collagenase, papain and lipase stir under conditions of pH7,
40 DEG C are cooled to after enzymatic hydrolysis 2h under the conditions of 40 DEG C -50 DEG C to continue to digest 1.5h, high temperature enzyme deactivation.
Preferably, the enzymatic hydrolysis condition are as follows: after Collagenase, papain carry out enzymatic hydrolysis 3h, high temperature enzyme deactivation；?
PH7, under the conditions of 40 DEG C to 50 DEG C, addition lipase digests 2h to 3.5h, stirs in the process, high temperature enzyme deactivation after enzymatic hydrolysis.
Preferably, the concentration is that nanofiltration is concentrated.
Preferably, the preparation method step includes:
(1) it thaws broken: after sea cucumber internal organ frozen goods at room temperature natural thaw, being crushed, obtain slurries；
(2) digest: complex enzyme, solid-liquid ratio 1:(3-4 will be added in slurries), it is digested；
(3) it is centrifuged: the system after enzymatic hydrolysis being centrifuged, supernatant is taken；
(4) nanofiltration is concentrated: by supernatant after aperture is 1 to 2nm nanofiltration membrane, it is concentrated into 30% or more mass concentration,
Obtain concentrate；
(5) dry: concentrate is spray-dried to obtain sea cucumber internal organ polypeptide solid product.
The present invention also provides a kind of sea cucumber internal organ polypeptide, such as above-mentioned preparation method is prepared.
The invention has the following advantages:
1. the flavor of sea cucumber internal organ polypeptide product can be significantly improved, including smell and mouthfeel.
The mouthfeel of the intrinsic penetrating odor of sea cucumber internal organ polypeptide product and bitter peppery mouth, the origin cause of formation are more complicated.This method
By lipase and protease compound action, system is changed particular by the use temperature of complex enzyme, dosage, using the time
The ingredient of lipid and its interactively with protein, polypeptide, and result in the hydrolysis and rearrangement of other ester bonds, glycosidic bond.It is logical
The verifying that naked eyes evaluation is tested is crossed, mouthfeel of the sea cucumber internal organ polypeptide by this method processing to penetrating odor and bitter peppery mouth
There is fairly obvious correcting, fishy smell reduces, and remains sea cucumber characteristic flavor on basis, significantly improves its acceptable degree, reduces
Using it as the difficulty of processing of the functional food of raw material and active constituent and health food, drug, its application range has been widened.
2. the color of sea cucumber internal organ polypeptide product can be effectively improved.
The sea cucumber internal organ polypeptide product of conventional method preparation is brown or crineous, and the product of this method preparation is shallow brown
Color or deep ecru.It is tested by naked eyes evaluation test with colorimetric detection, it was demonstrated that color improves significant.The improvement of color also improves
Acceptable degree and range is applied, reduced using it as the difficulty of processing of raw produce.
3. yield can be improved, the cost of product is reduced.
Compared with singly using the method for protease hydrolyzed, this method effectively increases yield.To improve flavor, it is usually taken
The technique that rinsing or blanching are carried out to raw material, water soluble ingredient loss, causes yield lower, and it is i.e. rectifiable that this technique is not required to rinsing
The bad flavor of product avoids the loss of active constituent.Test result shows that the yield of this technique is higher than conventional method, more
Higher than fulling.
Specific embodiment
The present invention relates to a kind of preparation method of sea cucumber internal organ polypeptide, step includes:
(1) it thaws broken: after sea cucumber internal organ frozen goods at room temperature natural thaw, being crushed, obtain slurries；It is described
Room temperature is no more than 20 DEG C；
(2) digest: complex enzyme, solid-liquid ratio 1:(3-4 will be added in slurries), it is digested；The complex enzyme is collagen
The combination of protease, papain and lipase；
(3) it is centrifuged: the system after enzymatic hydrolysis being centrifuged, supernatant is taken；The centrifugal condition are as follows: system is cooling
3000rpm is centrifuged 10min after to 30 DEG C, and taking supernatant, 6000rpm is centrifuged 10min again, takes supernatant again；
(4) nanofiltration is concentrated: by supernatant after aperture is 1 to 2nm nanofiltration membrane, it is concentrated into 30% or more mass concentration,
Obtain concentrate；
(5) dry: concentrate is spray-dried to obtain sea cucumber internal organ polypeptide solid product；The spray drying condition are as follows: into
180 DEG C of air temperature, 90 DEG C of leaving air temp, sample introduction speed 30ml/min.
As an embodiment of the present invention, ultrafiltration removing macromolecular is first carried out before spray drying step: being used
9000D filter membrane carries out ultrafiltration, removes macromolecular and impurity.
As an embodiment of the present invention, excessive hole resin removing small molecule is first carried out before nanofiltration concentration step:
Using D101 macroporous resin adsorption small molecular weight impurity, high-purity sea cucumber polypeptide is obtained；Meanwhile macroreticular resin being eluted and can get height
Concentration saponin(e and sea cucumber polysaccharide mixture.
Below with reference to embodiment, the present invention is further illustrated.
Instrument involved in following embodiments, reagent, material etc. are unless otherwise noted existing in the prior art
Conventional instrument, reagent, material etc., can be obtained by regular commercial sources.Experimental method involved in following embodiments, inspection
Survey method etc. is unless otherwise noted existing routine experiment method in the prior art, detection method etc..
Embodiment one:
The defrosting of 1.1 raw materials: weighing spring takes sea cucumber internal organ frozen goods 1kg, is statically placed in 20 DEG C of defrosting 3h.
1.2 historrhexis: homogenized is carried out using beater, 6000 turns/min of refiner is beaten 30s, obtains slurries.
It is crushed to improve enzymolysis efficiency.
1.3 enzymatic hydrolysis: complex enzyme, solid-liquid ratio 1:3, Collagenase 0.05%(2*10 will be added in slurries4U/g), pawpaw
Proteinase-10 .05%(2*104U/g), lipase 0.05%(1*104U/g), pH7 is stirred in the process, after 50 DEG C of enzymatic hydrolysis 3.5h, 90 DEG C
Act on the enzyme deactivation of 10min high temperature.
1.4 centrifugations: 3000rpm is centrifuged 10min after system is cooled to 30 DEG C, and taking supernatant, 6000rpm is centrifuged again
10min takes supernatant again.Centrifugation carries out in two times, improves centrifugal efficiency.
The concentration of 1.5 nanofiltration desalinations: system is concentrated into mass concentration by removing salt and small-molecule substance, nanofiltration membrane aperture 1nm
30% or more.
1.6 spray drying: 180 DEG C of inlet air temperature, 90 DEG C of leaving air temp, sample introduction speed 30ml/min.
Embodiment two:
The defrosting of 1.1 raw materials: weighing spring takes sea cucumber internal organ 1kg, is statically placed in 20 DEG C of defrosting 3h.
1.2 historrhexis: homogenized is carried out using beater, 6000 turns/min of refiner is beaten 30s.
1.3 enzymatic hydrolysis: solid-liquid ratio 1:3, Collagenase 0.05%(2*104U/g), papain 0.05%(2*104u/
G), lipase 0.05%(1*104U/g), pH7 is stirred in the process, cooled to after 50 DEG C of enzymatic hydrolysis 2h 40 DEG C continue digest 1.5h, 90
DEG C effect the enzyme deactivation of 10min high temperature.
1.4 centrifugations: 3000rpm is centrifuged 10min after system is cooled to 30 DEG C, and taking supernatant, 6000rpm is centrifuged again
10min takes supernatant again.
The concentration of 1.5 nanofiltration desalinations: system is concentrated into 30% or more mass concentration by nanofiltration membrane aperture 1nm.
1.6 spray drying；180 DEG C of inlet air temperature, 90 DEG C of leaving air temp, sample introduction speed 30ml/min.
Embodiment three:
The defrosting of 1.1 raw materials: weighing autumn takes sea cucumber internal organ 1kg, is statically placed in 10 DEG C of defrosting 5h.
1.2 historrhexis: homogenized is carried out using beater, 4000 turns/min of refiner is beaten 1min.
1.3 enzymatic hydrolysis: under the conditions of 1:4, pH7,50 DEG C of solid-liquid ratio, Collagenase 0.05%(2*104U/g), Papain
Enzyme 0.05%(2*104U/g) 90 DEG C of effect 10min high temperature enzyme deactivations after enzymatic hydrolysis 3h；Lipase 0.1% will be added in system after enzyme deactivation
(1*104U/g), pH7 digests 2h under the conditions of 40 DEG C, stirs in the process, 90 DEG C of effect 10min high temperature enzyme deactivations after enzymatic hydrolysis.
1.4 centrifugations: 3000rpm is centrifuged 15min after system is cooled to 30 DEG C, takes supernatant.
1.5 ultrafiltration remove macromolecular: carrying out ultrafiltration using 9000D filter membrane, remove macromolecular and impurity.
The concentration of 1.6 nanofiltration desalinations: system is concentrated into 30% or more mass concentration by nanofiltration membrane aperture 1nm.
1.7, which cross macroreticular resin, removes small molecule: using D101 macroporous resin adsorption small molecular weight impurity, obtains high-purity sea cucumber
Polypeptide；Meanwhile macroreticular resin being eluted and can get high concentration saponin(e and sea cucumber polysaccharide mixture.
1.8 spray drying: 160 DEG C of inlet air temperature, 90 DEG C of leaving air temp, sample introduction speed 20ml/min.
Test example
1 test method
The defrosting of 1.1 raw materials: weighing equivalent spring takes 4 parts of each 2kg of sea cucumber internal organ, is respectively labeled as 1,2,3, No. 4, quiet
It is placed in 20 DEG C of defrosting 3h.
1.2 pretreatments: No. 1 raw material is placed in 2L water to drainage after rinsing 10s, is in kind cleaned 3 times.2,3, No. 4
Raw material is without pretreatment.
1.3 historrhexis: homogenized, 6000 turns/min of refiner mashing are carried out to four parts of samples respectively using beater
30s。
1.4 enzymatic hydrolysis: pH7, protease and/or lipase, 50 DEG C of enzymatic hydrolysis 3.5h are stirred in the process, 90 DEG C of effect 10min high
Warm enzyme deactivation.
Wherein, 1, No. 2 sample only adds Collagenase 0.075%(2*104) and papain 0.075%(2* u/g
104U/g), No. 3 samples add Collagenase 0.05%(2*104U/g), papain 0.05%(2*104) and lipase u/g
0.05%(1*104U/g), No. 4 samples only add lipase 0.15%(1*104U/g).1,2,3 and No. 4 sample solid-liquid ratios are
1:3。
1.5 centrifugations: 3000rpm is centrifuged 10min after the system after 4 parts of enzymatic hydrolysis is cooled to 30 DEG C respectively, takes supernatant again
6000rpm is centrifuged 10min, takes supernatant again.
The concentration of 1.6 nanofiltration desalinations: system is concentrated into 30% or more mass concentration by nanofiltration membrane aperture 2nm.
1.7 spray drying；180 DEG C of inlet air temperature, 90 DEG C of leaving air temp, sample introduction speed 30ml/min.
The above are protease hydrolyzed, the comparison of the untreated protease hydrolyzed method of raw material after patented method and raw material cleaning are real
It tests, No. 1 (rinsing protease), No. 2 (protease), No. 3 (protease+lipase), No. 4 (lipase).
Flavor evaluation: flavor evaluation is carried out by naked eyes evaluation.It is molten that above-mentioned 4 parts of powdery products are configured to 1% respectively
Liquid randomly selects 15 professional technicians and 15 non-specialized-technical personnel and carries out respectively to the smell of four parts of solution and mouthfeel
Two number of components superposition calculation average marks are taken two effective digitals to obtain sensory scores by scoring.Standards of grading are shown in Table 1, scoring knot
Fruit is shown in Table 2.
2 sensory evaluation Score Lists of table
| No. 1 | No. 2 | No. 3 | No. 4 | |
| Smell score | 3.3 | 1.4 | 4.5 | 2.8 |
| Mouthfeel score | 2.4 | 1.8 | 4.2 | 2.5 |
| Total score | 5.7 | 3.2 | 8.7 | 5.3 |
By Analyses Methods for Sensory Evaluation Results it is found that smell and mouthfeel score that this method obtains product are obviously higher than conventional method,
It is also significantly better than fulling.The sea cucumber internal organ polypeptide handled by this method has ten to the mouthfeel of penetrating odor and bitter peppery mouth
Point apparent correcting, fishy smell reduces, and remains sea cucumber characteristic flavor on basis, significantly improves its acceptable degree, reduce with
It is the difficulty of processing of the functional food of raw material and active constituent and health food, drug, has widened its application range.
Color evaluation: being measured using color difference meter and color of the whiteness instrument to four parts of powdery products, and whiteness uses upper
The portable whiteness instrument measurement of the SBDY-2 of the happy elegant demeanour instruments and meters Co., Ltd in sea, other parameters use the limited public affairs of Hangzhou chromatogram science and technology
The distant place board CS-10 color difference meter of department measures.It chooses 4 different batches of product to be measured, every kind of product of each batch surveys 3 respectively
Group data, totally 12 groups of data are averaged, and the results are shown in Table 3.
3 whiteness instrument of table and colour difference meter test result table
| Whiteness | L (brightness) | a* | b* | |
| No. 1 | 10.7 | 61.70 | 3.49 | 11.25 |
| No. 2 | 7.3 | 50.84 | 2.43 | 14.33 |
| No. 3 | 31.3 | 76.02 | 5.69 | 20.20 |
| No. 4 | 12.8 | 63.33 | 2.18 | 15.96 |
15 professional technicians and 15 non-specialized-technical personnel are randomly selected respectively to the color of four parts of powdery products
Pool is scored, and average mark is calculated, and two effective digitals is taken to obtain color score.Standards of grading are shown in Table 4, and appraisal result is shown in Table 5.
The evaluation of 4 color of table requires table
The evaluation of 4 color of table requires table
5 color sensory evaluation Score Lists of table
| No. 1 | No. 2 | No. 3 | No. 4 | |
| Color score | 2.5 | 1.9 | 4.1 | 2.5 |
By table 3 it is found that this method, which obtains product, compares conventional method and fulling, whiteness, brightness are obviously improved, a*,
B* value is all larger than other two methods, illustrates the more inclined warm tones of tone.By table 5 it is found that the color sensory evaluation of this method obtains
Divide and is also apparently higher than first two method, it is corresponding with the result of table 3.Conventional method preparation sea cucumber internal organ polypeptide product be brown or
Crineous, and the product of this method preparation is deep ecru.The improvement of color also improves acceptable degree and using range, reduces
Using it as the difficulty of processing of raw produce.
Yield comparison: yield is calculated separately to four parts of powdery products according to formula, comparing result is shown in Table 6.
6 four parts of powdery product yield tables of table
| No. 1 | No. 2 | No. 3 | No. 4 | |
| Yield | 4.25% | 7.48% | 8.42% | 2.23% |
As shown in Table 6, the yield of this method is apparently higher than conventional method, in the premise for obtaining ideal flavor and appearance
Under, not only yield is higher than fulling, also above not rinsing the method for only using protease.The raising of yield may be digested with lipid
Promote disorganization, the destruction of lipoprotein promote proteolysis efficiency and non-aqueous ingredient emulsification it is related.It is single
Using, because of the participation of no exogenous protease, the reaction that sea cucumber visceral protein is degraded to polypeptide only relies in sea cucumber the case where lipase
The self-enzymatic hydrolysis of the dirty autolytic enzyme contained in itself, yield are lower.
Using judgment criteria such as above-mentioned smell, mouthfeel, whiteness, colors, embodiment one and two product of embodiment are surveyed
Examination, the results are shown in Table 7.
The comparison of table 7 embodiment one and two final products of embodiment
| Smell score | Mouthfeel score | Sense organ total score | Whiteness | Color score | Yield | |
| Embodiment one | 4.5 | 4.2 | 8.7 | 31.3 | 4.1 | 8.42% |
| Embodiment two | 4.5 | 4.5 | 9.0 | 35.8 | 4.3 | 8.20% |
The difference of embodiment one and embodiment two is to take cooling behaviour after the enzymolysis process of embodiment two proceeds to 2h
Make.The adjustment of hydrolysis temperature changes the action site and speed of action of lipase.By table 7 as it can be seen that embodiment two mouthfeel score,
It is better than embodiment one in whiteness, and reduces energy consumption.
Protease and lipase compound action and its process conditions are important for purposes of the invention.By protease and rouge
Fat enzyme compound action, occur while protein peptide chain being made to hydrolyze under conditions of not removing fat complicated esterlysis, esterification,
The reactions such as transesterification, alcoholysis, acidolysis, glycosidic bond dissociation, for example the dissociation and rearrangement, the bad sugar of flavor of the bad lipid of flavor occurs
The fatty acid that the dissociation of rouge and rearrangement, dissociation of other chemical bonds etc. are reacted, while being dissociateed is conducive to the emulsification of system, to not
Good flavor has embedding effect；Reaction can also promote the hydrophobic interaction of bitter peptides hydrophobic grouping.
The present invention can be summarized with others without prejudice to the concrete form of spirit or essential characteristics of the invention.Therefore, nothing
By from the point of view of which point, the embodiment above of the invention can only all be considered the description of the invention and cannot limit this hair
Bright, claims indicate the scope of the present invention, and above-mentioned explanation does not point out the scope of the present invention, therefore, with this
Any change in the comparable meaning and scope of claims of invention is all considered as including in claim of the invention
In the range of book.
Claims (7)
1. a kind of preparation method of sea cucumber internal organ polypeptide, step include: sea cucumber internal organ frozen goods are successively thawed, are digested,
Centrifugation, concentration, drying steps, which is characterized in that complex enzyme is used in the enzymolysis step, the complex enzyme is collagen
The combination of enzyme, papain and lipase；
The weight ratio of the Collagenase, papain and lipase is 1:1:(1-2).
2. preparation method as described in claim 1, which is characterized in that the enzymatic hydrolysis condition are as follows: Collagenase, Papain
Enzyme and lipase stir under conditions of pH7, after digesting 3.5h under the conditions of 40 DEG C -50 DEG C, high temperature enzyme deactivation.
3. preparation method as described in claim 1, which is characterized in that the enzymatic hydrolysis condition are as follows: Collagenase, Papain
Enzyme and lipase stir under conditions of pH7, cool to 40 DEG C after enzymatic hydrolysis 2h under the conditions of 40 DEG C -50 DEG C and continue to digest 1.5h, high
Warm enzyme deactivation.
4. preparation method as described in claim 1, which is characterized in that the enzymatic hydrolysis condition are as follows: Collagenase, Papain
After enzyme carries out enzymatic hydrolysis 3h, high temperature enzyme deactivation；Under the conditions of pH7,40 DEG C -50 DEG C, addition lipase digests 2h to 3.5h, stirs in the process
It mixes, high temperature enzyme deactivation after enzymatic hydrolysis.
5. such as the described in any item preparation methods of claim 2-4, which is characterized in that the concentration is that nanofiltration is concentrated.
6. preparation method as claimed in claim 5, which is characterized in that step includes:
(1) it thaws broken: after sea cucumber internal organ frozen goods at room temperature natural thaw, being crushed, obtain slurries；
(2) digest: complex enzyme, solid-liquid ratio 1:(3-4 will be added in slurries), it is digested；
(3) it is centrifuged: the system after enzymatic hydrolysis being centrifuged, supernatant is taken；
(4) nanofiltration is concentrated: by supernatant after aperture is 1 to 2nm nanofiltration membrane, being concentrated into 30% or more mass concentration, obtains
Concentrate；
(5) dry: concentrate is spray-dried to obtain sea cucumber internal organ polypeptide solid product.
7. a kind of sea cucumber internal organ polypeptide, which is characterized in that preparation method as claimed in claim 6 is prepared.
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