CN109897101A - A kind of collagen tripeptide and its production method and application - Google Patents
A kind of collagen tripeptide and its production method and application Download PDFInfo
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Abstract
The invention belongs to collagen tripeptide field, a kind of collagen tripeptide and its production method and application are disclosed.The production method of the collagen tripeptide include by fish scale through alkali cleaning, pickling and add water mill it is broken, heat treatment, high efficiency compounded enzyme preparate digests under the conditions of not adjusting pH value and not controlling temperature, inorganic nanofiltration ceramic membrane filter removal not readily dissolves the ingredient with the easy moisture absorption, high temperature high pressure process, film concentration and fluidized bed drying powder.It does not need to adjust enzymatic hydrolysis pH value using method provided by the invention, need that controlled enzymatic hydrolysis temperature, enzymolysis time are short, enzyme additive amount is few, enzymolysis product molecular weight is low and concentrate, and obtained collagen tripeptide powder water solubility is splendid and storage in be not easy the moisture absorption, which can be made into ordinary food, health food, dispensed food for baby, special medicine purposes formula food and beauty skin care product.
Description
Technical field
The invention belongs to collagen tripeptide field, in particular to a kind of collagen tripeptide and its production method and answer
With.
Background technique
The amino acid such as glycine, proline, the hydroxyproline that collagen is needed rich in human body.Collagen is a kind of high
Molecular functional protein, it is the chief component of skin, accounts for the specific gravity of dermal layer of the skin 80%, it is constituted in skin
One fine and closely woven elasticated net, securely locks moisture, supports skin.Collagen is the spiral fiber twisted by three peptide chains
Shape protein and the most abundant protein of people's in-vivo content, are distributed widely between connective tissue, skin, bone, gut cell
The positions such as matter and flesh chamber, ligament, sclera, account about 30% or more of human body gross protein, it rich in human body need proline,
The collagens characteristic amino acid such as hydroxyproline is important component in human body cell especially skin cell epimatrix.Glue
Former albumen is macro-molecular protein, and molecular weight cannot be directly absorbed by the body in 300,000Da or more.
Collagen tripeptide (Collagen tripeptide) is product of the collagen after enzyme hydrolysis, is collagen
Minimum unit, structure can be expressed as Gly-X-Y, be it is a kind of N-terminal have glycine high-purity tripeptides, molecular weight between
Between 200~500Da, average molecular weight is about 280Da, compared with the molecular weight of common collagen peptide 1000Da, collagen tripeptide without
Need to decompose can be absorbed and utilized by the body, and have many advantages, such as purity is high, no antigen, hypoallergenic.
CN103243144B discloses a kind of preparation method of collagen protein powder rich in collagen tripeptide, the preparation method with
Fish-skin first extracts to collagen and is degraded into small-molecular peptides using compound protease as raw material, recycles certainly
The Collagenase of main screening is degraded into collagen tripeptide to small-molecular peptides further orientation, then decolourize through centrifugation, deodorant,
Concentration, spray drying obtain the collagen protein powder for being rich in collagen tripeptide.
CN 101870995B discloses a kind of technique for preparing collagen tripeptide, including by fish-skin through a lye pre-treatment,
Primary denatured by boiling processing, the stepped combined hydrolysis of primary alkali protease, pancreatin, compound protease.Wherein, it is wanted after denatured by boiling
Residue is filtered off with gauze, and the hydrolyzate that three enzyme fractional hydrolysis obtain passes through inactivation, filtering, de- bitter, decoloration, vacuum concentration and spraying
It is dry that the collagen tripeptide that protein content is high, functional activity is good can be obtained.
However, problems and disadvantages existing for above-mentioned two pieces application for a patent for invention are as follows: the enzymolysis efficiency of the enzyme of use is low, enzymatic hydrolysis
Time is long, and enzyme additive amount is more, needs to adjust optimum pH and optimum temperature is digested, need stepwise discretization, complicated for operation, adjusts
Section peak enzymolysis-ability pH value can bring a large amount of inorganic ions into, and control peak enzymolysis-ability temperature energy consumption is larger；It needs using ultrafiltration membrane film mistake
Filter, ultrafiltration membrane role be retention macromolecular substances, film core used be organic material, not acid and alkali-resistance, non-refractory high pressure,
Flux is small, service life is short, and cleaning is inconvenient；The water solubility of prepared collagen tripeptide powder is poor, storage
In be easy the moisture absorption.
Summary of the invention
The purpose of the present invention is to provide one kind not to need to adjust enzymatic hydrolysis pH value, does not need controlled enzymatic hydrolysis temperature, enzymatic hydrolysis production
Object molecular weight is low and concentrates, film filtering only needs primary and easy to clean, the obtained collagen tripeptide powder water solubility of operation
The collagen tripeptide and its production method and application of the moisture absorption are not easy in splendid and storage.
Specifically, the present invention provides a kind of production methods of collagen tripeptide, method includes the following steps:
(1) raw material pre-treatment: fish scale is put into reaction kettle, and 2~10 times of fish scale weight of aqueous slkali, stirring and washing is added
1~12h is eluted with water, and 2~10 times of fish scale weight of acid solution is added later, and 1~12h of stirring and washing is eluted with water, and is added
3~5 times of water mills of fish scale weight are broken to be prepared into thick collagen solution；The mass-volume concentration of the aqueous slkali is 1~4%, institute
The mass-volume concentration for stating acid solution is 3~8%；
(2) heat: the thick collagen solution being heated into 20~40min at 90~110 DEG C, is cooled to later
50~55 DEG C, obtain denatured collagen solution；
(3) digest: it is the efficient of raw material weight 0.15~0.15% that dosage is added into the denatured collagen solution
Rate compounded enzyme preparate digests 0.5~2h in the case where not adjusting pH value, the natural conditions for not controlling temperature after mixing, and glue is made
Former three peptide solution of albumen；The high efficiency compounded enzyme preparate is by 25~30wt% compound protease, 25~30wt% basic protein
Enzyme, 15~20wt% pancreatin, 15~20wt% flavor protease, 7~8wt% animal proteolytic enzyme and 2~3wt% enzyme activition
Agent composition, the zymoexciter contain reducing agent and bivalent metal ion；
(4) inorganic nanofiltration ceramic membrane filter: use molecular cut off for the nothing of 800Da the collagen tripeptide solution
Machine nanofiltration ceramic membrane is filtered, and gained permeate is three peptide solution of high purity collagen；
(5) high temperature high pressure process: three peptide solution of high purity collagen is subjected to high temperature high pressure process, is handled
Liquid；
(6) it is concentrated: the treatment fluid being subjected to film concentration, obtains concentrate；
(7) dry powder processed: the concentrate is subjected to fluidized bed drying, obtains collagen tripeptide powder.
Further, the molar ratio of reducing agent and bivalent metal ion is 1:(0.5~2 in the zymoexciter).
Further, the reducing agent is dithiothreitol (DTT) and/or beta -mercaptoethanol.
Further, the bivalent metal ion is Mg2+、Zn2+And Ca2+Mixture.
Further, Mg2+、Zn2+And Ca2+Molar ratio be (0.9~1.1): (0.9~1.1): 1.
Further, the filter core of the inorganic nanofiltration ceramic membrane is using in siliceous, aluminum, magnesia, sa, simatic
Any one material is made.
Further, the condition of the high temperature high pressure process include temperature be 110~130 DEG C, pressure be 0.1~
0.3MPa, time are 5~20min.
The present invention also provides the collagen tripeptides produced by the above method.
In addition, the present invention also provides the collagen tripeptides as ordinary food, health food, baby formula
The application of food, special medicine purposes formula food and beauty skin care product raw materials for production.
Key means of the invention are using specific raw material pre-treatment, heat treatment and specific high efficiency compound enzyme
Preparation enzymatic hydrolysis.Wherein, fish scale is successively used aqueous slkali and acid solution to impregnate and can not only clean, but also fish scale can be made to steep
Hair water suction, makes it easier to subsequent enzymatic hydrolysis.The heat treatment enables to collagen to be denaturalized, and makes it easier for being digested, mention
High enzymolysis efficiency.The high efficiency compounded enzyme preparate is by 25~30wt% compound protease, 25~30wt% alkali protease, 15
~20wt% pancreatin, 15~20wt% flavor protease, 7~8wt% animal proteolytic enzyme and 2~3wt% zymoexciter are (also
The mixture of former agent and bivalent metal ion) composition, this specific proteinase synergy, which cooperates, to be very beneficial for scale collagen
Albumen is digested, and efficiently specificity can digest fish scale collagen, and reducing agent contained in zymoexciter and divalent are golden
Category ion can activate the enzyme activity of these types of protease, by several protease above in the work of reducing agent and bivalent metal ion
There is the enzyme preparation for compounding formation according to the above ratio under extremely strong collagen to digest efficiency.Using the above highly efficiency compositional enzyme
Preparation digests denatured collagen, in the case where not adjusting pH value, not controlling the natural conditions of temperature, is adding a small amount of albumen
The purpose of thorough collagen protein enzymolysis can be realized under the premise of enzyme, enzymolysis product molecular weight is small and concentrates, and enzymolysis product is free
Amino acid content is low, and molecular weight is that 1000Da collagen tripeptide ratio below can reach 100%, between 200~500Da
Collagen tripeptide ratio can reach 85% or more.
To sum up, beneficial effects of the present invention are as follows:
(1) the combination technique means digested using specific raw material pre-treatment, heat treatment and high efficiency compounded enzyme preparate,
The enzymolysis efficiency of collagen protein enzymolysis is significantly improved, so that in the natural item for not adjusting optimum pH, not controlling optimum temperature
Under part, add a small amount of protease, digest the short period, can thorough collagen protein enzymolysis, enzymolysis product molecular weight is small and concentrates,
Enzymolysis product free aminoacid content is low, and molecular weight is that 1000Da collagen tripeptide ratio below can reach 100%, is situated between
85% or more can be reached in the collagen tripeptide ratio of 200~500Da.In addition, not needing in enzymolysis process using acid
Alkali adjusts peak enzymolysis-ability pH value, does not bring inorganic ions into, subsequent not need that nanofiltration membrane is again taken to remove inorganic ions, simplifies work
Skill step；In enzymolysis process, do not need yet adjust peak enzymolysis-ability temperature, compared with the prior art, it is not only easy to operate but also
Save energy consumption.
(2) technological means that inorganic nanofiltration ceramic membrane filter is used after enzymatic hydrolysis removes the substance that not readily dissolves and easily
The carbohydrate content of the moisture absorption improves the water solubility of finally obtained collagen tripeptide finished product, and during finished stock storage not
Easy moisture absorption agglomeration.Inorganic nanofiltration ceramic membrane has organic solvent-resistant, high temperature resistant, acid and alkali-resistance, machinery strong compared with organic nanofiltration membrane
The advantage that degree is high, flux is big and the service life is long.The present invention need to only use a nanofiltration membrane treatment, than both needing to retain using ultrafiltration membrane
Macromolecular substances need to compare using the prior art of nanofiltration membrane desalination and concentration again, significantly simplify processing step, and at film
Reason effect is different.
(3) gained collagen tripeptide is used into high temperature high pressure process, such energy after inorganic nanofiltration ceramic membrane filter
The water solubility of finally obtained collagen tripeptide finished product is enough further increased, finished product is finer and smoother, is highly soluble in water.
Specific embodiment
The present invention described in detail below.
In the present invention, initial feed fish scale can be marine fishes fish scale, or freshwater fish fish scale.
In the present invention, the purpose of alkali cleaning is to remove fat and foreign protein.The purpose of pickling is in order to enable fish scale
Be soaked water suction, makes it easier to subsequent enzymatic hydrolysis.The mass-volume concentration of aqueous slkali employed in the raw material pre-treatment step is 1
~4%, the mass-volume concentration of the acid solution is 3~8%." mass-volume concentration " (w/v%) described herein refers to
The grams of contained solute in 100ml solution, that is, the mass-volume concentration of the aqueous slkali refers to 100ml aqueous slkali for 1~4%
In contain 1~4g alkali, similarly, the mass-volume concentration of the acid solution is 3~8% to refer to and contain 3~8g in 100ml acid solution
Acid.
In the present invention, the effect of the heat treatment be so that collagen be denaturalized, be easier to after heat-treated by
Enzymatic hydrolysis, can be improved enzymolysis efficiency.
In the present invention, the enzymatic hydrolysis carries out under the conditions of not adjusting pH value, not controlling temperature.After the enzymatic hydrolysis, from
It is extracted in fish scale and the collagen after heated denaturation can be degraded into that free aminoacid content is low, molecular weight is 1000Da's
Collagen tripeptide ratio can reach 100% (between the collagen tripeptide ratio of 200~500Da can reach 85% with
On) enzymolysis product.Wherein, protease contained in the high efficiency compounded enzyme preparate can pass through commercially available acquisition.For example,
The compound protease can be the compound protease purchased from Novozymes Company.The alkali protease can be for purchased from Guang Xinan
Ning Pangbo bioengineering Co., Ltd is originated from the resulting alkali protease of the lichen bacillus ferments.The pancreatin can be purchase
Resulting pancreatin is naturally extracted from pig, ox pancreas from Nanning Pang Bo bioengineering Co., Ltd.The flavor albumen
Enzyme can be to be originated from the resulting flavor protease of aspergillus oryzae fermentation purchased from Nanning Pang Bo bioengineering Co., Ltd.It is described
Animal proteolytic enzyme can be the animal protein water from complex enzyme purchased from Nanning Pang Bo bioengineering Co., Ltd
Solve enzyme.
The zymoexciter contains reducing agent and bivalent metal ion.Wherein, the reducing agent is preferably dithiothreitol (DTT)
And/or beta -mercaptoethanol.The bivalent metal ion is preferably selected from Mg2+、Zn2+And Ca2+At least one of, more preferably Mg2 +、Zn2+And Ca2+Mixture, most preferably Mg2+、Zn2+And Ca2+According to molar ratio (0.9~1.1): (0.9~1.1): 1 compounding
Obtained mixture.The bivalent metal ion can exist in the form of the salt containing the above metal ion.It is particularly preferred that
The zymoexciter is by reducing agent and bivalent metal ion and can be with the anion group of the bivalent metal ion forming salt
At.In addition, the molar ratio of reducing agent and bivalent metal ion is particularly preferably 1:(0.5~2 in the zymoexciter).
In the present invention, the effect of the ceramic membrane filter be remove the carbohydrate of the ingredient not readily dissolved and the easy moisture absorption at
Point, the water solubility of finally obtained collagen tripeptide finished product is improved, and moisture absorption agglomeration is not easy during finished stock storage.Nothing
Machine nanofiltration ceramic membrane compared with organic nanofiltration membrane, have organic solvent-resistant, high temperature resistant, acid and alkali-resistance, high mechanical strength, flux big and
The advantage of service life length.The filter core of the inorganic nanofiltration ceramic membrane preferably uses in siliceous, aluminum, magnesia, sa and simatic
Any one material be made.The molecular cut off of the inorganic nanofiltration ceramic membrane is 800Da, when molecular cut off is more than
When 800Da, the carbohydrate content for removing the ingredient not readily dissolved and the easy moisture absorption is not thorough, when molecular cut off is less than 800Da, glue
Former albumen tripeptides loss late is high.
In the present invention, the effect of the high temperature high pressure process be further increase finally obtained collagen tripeptide at
The water solubility of product easily dissolves so that finished product is finer and smoother, and three peptide solution of high purity collagen is by high temperature and pressure
After reason, it can also play the role of thoroughly sterilizing and eliminating hypersensitive.Preferably, the high temperature high pressure process is not less than in temperature
110 DEG C, pressure is not less than carrying out under conditions of 0.1MPa；It is highly preferred that the condition of the high temperature high pressure process includes that temperature is
110~130 DEG C, pressure is 0.1~0.3MPa, and the time is 5~20min.
In the present invention, the effect of the film concentration is by three peptide solution of high purity collagen after high temperature high pressure process
In moist part or all removal.
In the present invention, the effect of the fluidized bed drying is that concentrate is processed into powder.
In addition, can also be packed, after formation collagen tripeptide powder to be dried to sell outside.
Below with reference to embodiment, the content of the present invention will be explained in more detail.It should be appreciated that the embodiment of the present invention is example
Property, it is intended to it is used to explain the present invention, and is not considered as limiting the invention.Particular technique or item are not specified in embodiment
Part person described technology or conditions or carries out according to the literature in the art according to product description.Agents useful for same or instrument
Production firm person is not specified in device, and being can be with conventional products that are commercially available.
In following embodiment and comparative example, the compound protease, alkali protease, pancreatin, flavor protease, animal egg
The source of white hydrolase and manufacturing enterprise are as shown in table 1 below:
Table 1
| The title of enzyme | Source | Manufacturing enterprise |
| Compound protease | Complex enzyme | Novozymes Company |
| Alkali protease | The lichen bacillus ferments | Nanning Pang Bo bioengineering Co., Ltd |
| Pancreatin | Pig, ox pancreas naturally extract | Nanning Pang Bo bioengineering Co., Ltd |
| Flavor protease | Aspergillus oryzae fermentation | Nanning Pang Bo bioengineering Co., Ltd |
| Animal proteolytic enzyme | Complex enzyme | Nanning Pang Bo bioengineering Co., Ltd |
Embodiment 1
(1) raw material pre-treatment: marine fishes (Larimichthys crocea) fish scale is put into reaction kettle, and 2 times of fish scale weight of alkali is added
Solution (sodium hydroxide solution, similarly hereinafter), stirring and washing 12h are eluted with water, later with removing the fat and other impurities of fish scale
2 times of fish scale weight of acid solution (hydrochloric acid solution, similarly hereinafter), stirring and washing 12h is added, then is washed with deionized water, fish scale is added
3 times of water mills of weight are broken to be prepared into thick collagen solution.Wherein, the mass-volume concentration of aqueous slkali is 4%, the quality of acid solution
Volumetric concentration is 3%.
(2) heat: thick collagen solution being heated into 30min under 100 DEG C of hot water, is cooled to 50~55 later
DEG C, obtain denatured collagen solution.
(3) it digests: the high efficiency compound enzyme of 0.1% (w/w) raw material weight being added into the denatured collagen solution
Preparation digests 1h after mixing, and collagen tripeptide solution is made.The high efficiency compounded enzyme preparate is by the compound egg of 25wt%
White enzyme, 30wt% alkali protease, 15wt% pancreatin, 20wt% flavor protease, 7wt% animal proteolytic enzyme and 3wt%
Zymoexciter composition.Wherein, the zymoexciter is by reducing agent beta -mercaptoethanol and bivalent metal ion Mg2+、Zn2+、Ca2+Group
At the molar ratio of the reducing agent and bivalent metal ion is 1:2, the bivalent metal ion Mg2+、Zn2+And Ca2+Mole
Than for 1:1:1.
(4) inorganic nanofiltration ceramic membrane filter: use molecular cut off for the nothing of 800Da the collagen tripeptide solution
Machine nanofiltration ceramic membrane is filtered, and gained permeate is three peptide solution of high purity collagen.
(5) high temperature high pressure process: by three peptide solution of high purity collagen temperature is 130 DEG C, pressure is
5min is handled under conditions of 0.1MPa, obtains treatment fluid.
(6) it is concentrated: treatment fluid being subjected to film concentration, obtains concentrate.
(7) dry powder processed and packaging: carrying out fluidized bed drying powder for concentrate, after packaging collagen tripeptide at
Product.
Embodiment 2
(1) raw material pre-treatment: freshwater fish (grass carp) fish scale is put into reaction kettle, and 10 times of fish scale weight of alkali soluble is added
Liquid, stirring and washing 1h are eluted with water with removing the fat and other impurities of fish scale, and it is molten that 10 times of fish scale weight of acid is added later
Liquid, stirring and washing 1h, then be washed with deionized water are added that 4 times of water mills of fish scale weight are broken to be prepared into thick collagen solution.Its
In, the mass-volume concentration of aqueous slkali is 1%, and the mass-volume concentration of acid solution is 8%.
(2) heat: thick collagen solution being heated into 30min under 100 DEG C of hot water, is cooled to 50~55 later
DEG C, obtain denatured collagen solution；
(3) it digests: the high efficiency compound enzyme of 0.1% (w/w) raw material weight being added into the denatured collagen solution
Preparation digests 1h after mixing, and collagen tripeptide solution is made.The high efficiency compounded enzyme preparate is by the compound egg of 30wt%
White enzyme, 25wt% alkali protease, 20wt% pancreatin, 15wt% flavor protease, 8wt% animal proteolytic enzyme, 2wt% enzyme
Activator composition, wherein the zymoexciter is by reducing agent dithiothreitol and bivalent metal ion Mg2+、Zn2+、Ca2+Compounding
The molar ratio of composition, the reducing agent and bivalent metal ion is 1:0.5, the bivalent metal ion Mg2+、Zn2+And Ca2+'s
Molar ratio is 1:1:1.
(4) inorganic nanofiltration ceramic membrane filter: use molecular cut off for the nothing of 800Da the collagen tripeptide solution
Machine nanofiltration ceramic membrane is filtered, and gained permeate is three peptide solution of high purity collagen.
(5) high temperature high pressure process: by three peptide solution of high purity collagen temperature is 120 DEG C, pressure is
10min is handled under conditions of 0.2MPa, obtains treatment fluid.
(6) it is concentrated: treatment fluid being subjected to film concentration, obtains concentrate.
(7) dry powder processed and packaging: carrying out fluidized bed drying powder for concentrate, after packaging collagen tripeptide at
Product.
Embodiment 3
(1) raw material pre-treatment: freshwater fish (crucian) fish scale is put into reaction kettle, and 6 times of fish scale weight of alkali soluble is added
Liquid, stirring and washing 6h are eluted with water with removing the fat and other impurities of fish scale, and it is molten that 6 times of fish scale weight of acid is added later
Liquid, stirring and washing 6h, then be washed with deionized water are added that 5 times of water mills of fish scale weight are broken to be prepared into thick collagen solution.Its
In, the mass-volume concentration of aqueous slkali is 2%, and the mass-volume concentration of acid solution is 5%.
(2) heat: thick collagen solution being heated into 30min under 100 DEG C of hot water, is cooled to 50~55 later
DEG C, obtain denatured collagen solution；
(3) it digests: the high efficiency compound enzyme of 0.1% (w/w) raw material weight being added into the denatured collagen solution
Preparation digests 1h after mixing, and collagen tripeptide solution is made.The high efficiency compounded enzyme preparate is compound by 27.5wt%
Protease, 27.5wt% alkali protease, 17.5wt% pancreatin, 17.5wt% flavor protease, the hydrolysis of 7.5wt% animal protein
Enzyme and 2.5wt% zymoexciter composition, wherein the zymoexciter is by reducing agent dithiothreitol, beta -mercaptoethanol and two
Valence metal ion Mg2+、Zn2+、Ca2+It forms, the molar ratio of dithiothreitol (DTT) and beta -mercaptoethanol is 1:1, institute in the reducing agent
The molar ratio for stating reducing agent and bivalent metal ion is 1:1, the bivalent metal ion Mg2+、Zn2+And Ca2+Molar ratio be 1:
1:1。
(4) inorganic nanofiltration ceramic membrane filter: use molecular cut off for the nothing of 800Da the collagen tripeptide solution
Machine nanofiltration ceramic membrane is filtered, and gained permeate is three peptide solution of high purity collagen.
(5) high temperature high pressure process: by three peptide solution of high purity collagen temperature is 110 DEG C, pressure is
20min is handled under conditions of 0.3MPa, obtains treatment fluid.
(6) it is concentrated: treatment fluid being subjected to film concentration, obtains concentrate.
(7) dry powder processed and packaging: concentrate carries out fluidized bed drying powder, obtains collagen tripeptide finished product after packaging.
Comparative example 1
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Compound protease in enzyme preparation is substituted using the animal proteolytic enzyme of identical weight percentage composition, remaining and 1 phase of embodiment
Together, reference collagen tripeptide finished product is obtained.
Comparative example 2
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Alkali protease in enzyme preparation is substituted using the animal proteolytic enzyme of identical weight percentage composition, remaining and 1 phase of embodiment
Together, reference collagen tripeptide finished product is obtained.
Comparative example 3
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Pancreatin in enzyme preparation is substituted using the animal proteolytic enzyme of identical weight percentage composition, remaining is same as Example 1, obtains
Reference collagen tripeptide finished product.
Comparative example 4
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Flavor protease in enzyme preparation is substituted using the animal proteolytic enzyme of identical weight percentage composition, remaining and 1 phase of embodiment
Together, reference collagen tripeptide finished product is obtained.
Comparative example 5
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Animal proteolytic enzyme in enzyme preparation uses the basic protein enzymes extraction of identical weight percentage composition, remaining and 1 phase of embodiment
Together, reference collagen tripeptide finished product is obtained.
Comparative example 6
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency compound enzyme
Zymoexciter contained in preparation is completely by bivalent metal ion Mg2+、Zn2+、Ca2+It is 1:1:1 compounding composition according to molar ratio,
Remaining is same as Example 1, obtains reference collagen tripeptide finished product.
Comparative example 7
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency compound enzyme
Zymoexciter contained in preparation is made of beta -mercaptoethanol completely, remaining is same as Example 1, obtains reference collagen three
Peptide finished product.
Comparative example 8
Collagen tripeptide is produced according to the method for embodiment 1, unlike, in enzymolysis process, high efficiency is compounded
Zymoexciter contained in enzyme preparation is substituted using the animal proteolytic enzyme of identical weight percentage composition, remaining and embodiment 1
It is identical, obtain reference collagen tripeptide finished product.
Test case
(1) high efficiency compounded enzyme preparate hydrolysis result detects:
In Examples 1 to 3 and comparative example 1~8, marine fishes fish scale or freshwater fish fish scale, by raw material pre-treatment,
After heat treatment, digested under the conditions of not adjusting pH value, not controlling temperature using high efficiency compounded enzyme preparate, in order to assess
Hydrolysis result, after enzymatic hydrolysis, using the method in " National Standard of the People's Republic of China GB/T 22729-2008 " appendix A
Enzymolysis product detection is carried out, as a result as shown in table 2 below:
Table 2
As shown in Table 2, the enzymolysis product molecular weight of Examples 1 to 3 is small and concentrates, enzymolysis product free amino acid
Content is low (ratio less than 200Da is less than 5%), and collagen tripeptide ratio of the molecular weight less than 1000Da has reached 100%,
Reach 85% or more between the collagen tripeptide ratio of 200~500Da, enzymolysis product molecular weight is substantially better than comparative example 1
~8.It is possible thereby to illustrate, the combination skill digested using specific raw material pre-treatment, heat treatment and high efficiency compounded enzyme preparate
Art means can significantly improve the enzymolysis efficiency of collagen protein enzymolysis, so that not adjusting optimum pH, not controlling optimum temperature
Natural conditions under, add a small amount of protease, digest the short period, can thorough collagen protein enzymolysis.
(2) collagen tripeptide finished product physical and chemical index detects:
Physical and chemical index detection is carried out to the collagen tripeptide finished powder that Examples 1 to 3 obtains, wherein hydroxyproline
Detection method is carried out referring to GB/T 9695.23, and total nitrogen detection method is carried out referring to GB 5009.5, ash content detection method reference
GB5009.4 is carried out, as a result as shown in table 3 below:
Table 3
As shown in Table 3, hydroxyproline, total nitrogen, the ash content for the collagen tripeptide finished product that Examples 1 to 3 obtains
Index is significantly better than the requirement of " national food safety standard collagen peptide GB 31645-2018 ", illustrates using the present invention
The collagen tripeptide purity is high of the method production of offer, quality is good, and especially content of ashes only has 0.8~1.3g/100g.It says
It is bright to be digested using this specific high efficiency compounded enzyme preparate of the present invention in the case where not adjusting pH value condition, it does not need to use soda acid
Finished product content of ashes can be greatly reduced to will not introduce inorganic ions in production process by adjusting pH value.
(3) light transmittance of the water solubility of collagen tripeptide finished product and collagen tripeptide solution detects:
The collagen tripeptide finished powder 8g that Examples 1 to 3 obtains is poured into different cups respectively, with 60 DEG C of 300mL
Left and right warm water reconstitutes, and i.e. all dissolutions, obtain collagen tripeptide lysate in the 1s time, and it is saturating to visually observe lysate clarification
It is bright, without any precipitating.The light transmittance of several collagen tripeptide lysates is detected at 620nm using ultraviolet specrophotometer,
As a result as shown in table 4 below:
Table 4
| Project | Light transmittance (%) |
| Embodiment 1 | 95 |
| Embodiment 2 | 96 |
| Embodiment 3 | 95 |
As shown in Table 4, illustrate the technological means using inorganic nanofiltration ceramic membrane filter, remove the object not readily dissolved
Matter, then the technological means of the high temperature high pressure process using peptide solution, further increase the solubility property of finished product, so that the glue obtained
Former albumen tripeptides finished product is highly soluble in that water, water solubility are splendid, lysate clear after being dissolved in water, without any precipitating, thoroughly
Light rate is high, when consumer brews this product, without stirring for it is the technology that the prior art is not achieved that convenient, edible experience is good
Effect.
(4) the shelf life test of collagen tripeptide finished product:
The collagen tripeptide finished powder 8g that Examples 1 to 3 obtains is poured into packaging bag respectively, room temperature opening is placed
There is the time agglomerated on indoor Display Rack, visually observing finished product, as a result as shown in table 5 below:
Table 5
| Project | There is the time (day) agglomerated |
| Embodiment 1 | 66 |
| Embodiment 2 | 59 |
| Embodiment 3 | 62 |
The collagen tripeptide of prior art production is easy moisture absorption agglomeration, influences the sale of product in storage.By
The result of table 5 it is found that the present invention uses the technological means of inorganic nanofiltration ceramic membrane filter, can remove the carbohydrate of the easy moisture absorption at
Point, so that being not easy moisture absorption agglomeration during finished stock storage, it is easy to store, is the technical effect that the prior art is not achieved.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art are not departing from the principle of the present invention and objective
In the case where can make changes, modifications, alterations, and variations to the above described embodiments within the scope of the invention.
Claims (9)
1. a kind of production method of collagen tripeptide, which is characterized in that method includes the following steps:
(1) raw material pre-treatment: fish scale is put into reaction kettle, the aqueous slkali of 2~10 times of fish scale weight of addition, and stirring and washing 1~
12h is eluted with water, and 2~10 times of fish scale weight of acid solution is added later, and 1~12h of stirring and washing is eluted with water, and fish scale is added
3~5 times of water mills of weight are broken to be prepared into thick collagen solution；The mass-volume concentration of the aqueous slkali is 1~4%, the acid
The mass-volume concentration of solution is 3~8%；
(2) heat: the thick collagen solution is heated into 20~40min at 90~110 DEG C, it is cooled to 50 later~
55 DEG C, obtain denatured collagen solution；
(3) it digests: it is multiple that the high efficiency that dosage is raw material weight 0.15~0.15% being added into the denatured collagen solution
With enzyme preparation, 0.5~2h is digested in the case where not adjusting pH value, the natural conditions for not controlling temperature after mixing, collagen egg is made
White three peptide solution；The high efficiency compounded enzyme preparate is by 25~30wt% compound protease, 25~30wt% alkali protease, 15
~20wt% pancreatin, 15~20wt% flavor protease, 7~8wt% animal proteolytic enzyme and 2~3wt% zymoexciter group
At the zymoexciter contains reducing agent and bivalent metal ion；
(4) inorganic nanofiltration ceramic membrane filter: molecular cut off is used to receive for the inorganic of 800Da the collagen tripeptide solution
Filter ceramic membrane is filtered, and gained permeate is three peptide solution of high purity collagen；
(5) high temperature high pressure process: three peptide solution of high purity collagen is subjected to high temperature high pressure process, obtains treatment fluid；
(6) it is concentrated: the treatment fluid being subjected to film concentration, obtains concentrate；
(7) dry powder processed: the concentrate is subjected to fluidized bed drying, obtains collagen tripeptide powder.
2. the production method of collagen tripeptide according to claim 1, which is characterized in that restored in the zymoexciter
The molar ratio of agent and bivalent metal ion is 1:(0.5~2).
3. the production method of collagen tripeptide according to claim 1, which is characterized in that the reducing agent is two sulphur Soviet Union
Sugar alcohol and/or beta -mercaptoethanol.
4. the production method of collagen tripeptide according to claim 1, which is characterized in that the bivalent metal ion is
Mg2+、Zn2+And Ca2+Mixture.
5. the production method of collagen tripeptide according to claim 4, which is characterized in that Mg2+、Zn2+And Ca2+Rub
You are than being (0.9~1.1): (0.9~1.1): 1.
6. the production method of collagen tripeptide described according to claim 1~any one of 5, which is characterized in that described
The filter core of inorganic nanofiltration ceramic membrane is made using any one material in siliceous, aluminum, magnesia, sa, simatic.
7. the production method of collagen tripeptide described according to claim 1~any one of 5, which is characterized in that described
The condition of high temperature high pressure process includes that temperature is 110~130 DEG C, and pressure is 0.1~0.3MPa, and the time is 5~20min.
8. the collagen tripeptide that the method as described in any one of claim 1~7 produces.
9. collagen tripeptide according to any one of claims 8 is as ordinary food, health food, dispensed food for baby, special
The application of medical usage formula food and beauty skin care product raw materials for production.
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| CN116769861A (en) * | 2023-05-23 | 2023-09-19 | 纳泰(阳江)生物科技有限公司 | Collagen tripeptide with high GPH content and preparation method thereof |
| CN117165646A (en) * | 2023-08-25 | 2023-12-05 | 广州菲勒生物科技有限公司 | Collagen tripeptide composition and purification method thereof |
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