CN108467487A - The dextrin modified casein polypeptide conjugate of starch base, preparation method and application - Google Patents
The dextrin modified casein polypeptide conjugate of starch base, preparation method and application Download PDFInfo
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- CN108467487A CN108467487A CN201810286656.2A CN201810286656A CN108467487A CN 108467487 A CN108467487 A CN 108467487A CN 201810286656 A CN201810286656 A CN 201810286656A CN 108467487 A CN108467487 A CN 108467487A
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- dextrin
- starch base
- casein polypeptide
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G81/00—Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
Abstract
The invention discloses a kind of dextrin modified casein polypeptide conjugate of starch base, preparation method and applications.The preparation method of the dextrin modified casein polypeptide conjugate of starch base includes：So that casein polypeptide and starch base dextrin is carried out Mei Lade covalence graft reactions, forms the dextrin modified casein polypeptide conjugate of starch base.Using high-pressure homogeneous and micro jetting technology, so that the aqueous phase solution of the dextrin modified casein polypeptide conjugate of the starch base is uniformly mixed with emulsification oil phase component, obtain the dextrin modified casein polypeptide nanoemulsions of starch base of oil-in-water type.The small drop sizes of nanoemulsions prepared by the present invention allow high-efficiency delivery, hydrophobic bioactive material absorbing efficiency can be improved, have the advantages that preferable antioxidant activity, antibiotic property, preferable emulsibility and emulsion stability, there is extensive use in the fields such as stabilization of the delivering of nutriment, the controlled release of drug, bioactive substance.
Description
Technical field
The present invention relates to a kind of preparation of the dextrin modified casein polypeptide nanoemulsions of starch base more particularly to a kind of starch
Dextrin modified casein polypeptide conjugate of base and preparation method thereof and the dextrin modified casein polypeptide conjugate system of the starch base
The nanoemulsions and its application that the casein polypeptide that standby dextrin covalence graft is modified is stablized belong to albumen deep process technology neck
Domain.
Background technology
Nanoemulsions are a kind of colloidal dispersion systems, are non-thermodynamic stable systems, transparent or translucent, granularity ruler
The very little dynamic stabilization characteristic between 50-200nm, analysed with anti-settling and breast.Oil-in-water type nano-emulsion liquid is insoluble as water
Property bioactive substance carrier, should have good emulsibility and emulsion stability, and efficient delivering effect, therefore breast
The selection and application of agent are paid attention to by more and more people.
Casein has the characteristics such as high biocompatibility, biological degradability, biological safety as emulsifier.Through ultrasound
Enzyme process moderately hydrolysis casein peptide obtained is assisted, so that its functional group is preferably exposed, improves its emulsification property.Junket
Protein peptides are modified through starch base dextrin covalence graft again, and solubility increases, emulsibility and emulsion stability enhancing, inoxidizability
It can improve, antibacterial ability has a degree of improvement, when as delivery vector, further improves the change of embedding active material
Learn stability.Therefore, nanoemulsions are prepared as bioactive substance carrier as emulsifier, there is certain research meaning
Justice.
Therefore the hot spot for stablizing effective stabilizer for preparing oil-in-water type nano-emulsion liquid as research how is prepared
Project, and industry research staff's urgent problem to be solved at present.
Invention content
The main purpose of the present invention is to provide a kind of dextrin modified casein polypeptide conjugate of starch base and its preparation sides
Method, with overcome the deficiencies in the prior art.
Another main purpose of the present invention is to provide based on the dextrin modified casein polypeptide conjugate system of aforementioned starch base
Dextrin modified casein polypeptide nanoemulsions of standby starch base and preparation method thereof.
Another main purpose of the present invention is to provide answering for the dextrin modified casein polypeptide nanoemulsions of aforementioned starch base
With.
For realization aforementioned invention purpose, the technical solution adopted by the present invention includes：
An embodiment of the present invention provides a kind of preparation method of the dextrin modified casein polypeptide conjugate of starch base, packets
It includes：
Casein polypeptide and starch base dextrin are provided；
Make the mixed system comprising the casein polypeptide and starch base dextrin carry out Mei Lade in 80~120 DEG C covalently to connect
Branch 90~120min of reaction, forms the dextrin modified casein polypeptide conjugate of starch base.
In some embodiments, the mass ratio of the casein polypeptide and starch base dextrin is 1：1~2.
In some embodiments, the preparation method includes：The casein polypeptide is dissolved in phosphate buffer, it
After the starch base dextrin is added.
Further, the preparation method further includes：After the completion of the Mei Lade covalence grafts react, by the reaction
System cools down, and freeze-drying is to -20 DEG C or less later.
Further, the preparation method includes：Under conditions of pH value is 6.8~7.5, handled using ultrasonic wave added,
So that casein and trypsase is carried out 90~120min of enzymatic hydrolysis reaction in 40~60 DEG C, obtains casein polypeptide.
Further, the degree of hydrolysis of the casein polypeptide is 10~20%.
In some embodiments, the preparation method includes：Waxy corn starch Heat Gelatinization is set to be cooled to 40~60 DEG C, it
Pullulanase is added afterwards and carries out de- branch processing 20~for 24 hours, obtains dextrin solution, then using polyethylene glycol gradually the precipitation method by dextrin
Solution obtains the uniform starch base dextrin of the degree of polymerization in 20~25 DEG C of water-baths 20~be classified for 24 hours.
Further, the degree of polymerization of the starch base dextrin is 70~100.
The embodiment of the present invention additionally provides the dextrin modified casein polypeptide conjugate of starch base prepared by preceding method.
The embodiment of the present invention additionally provides a kind of preparation method of the dextrin modified casein polypeptide nanoemulsions of starch base,
Including：
The dextrin modified casein polypeptide conjugate of starch base is prepared according to preceding method；
Using high-pressure homogeneous and micro jetting technology, keep the water phase of the dextrin modified casein polypeptide conjugate of the starch base molten
Liquid is uniformly mixed with emulsification oil phase component, obtains the dextrin modified casein polypeptide nanoemulsions of starch base of oil-in-water type.
In some embodiments, the preparation method specifically includes：Using high pressure homogenization technique, to the emulsification oil phase group
Divide and the mixed liquor of aqueous phase solution carries out 2~5min of shearing in 15 DEG C or less, forms thick lotion, use high pressure microjet skill later
The thick lotion is further homogenized processing by art at 100~120MPa, obtains the dextrin modified casein polypeptide of starch base
Nanoemulsions store at 2~6 DEG C later.
Further, the preparation method includes：So that hydrophobic bioactive substance is dissolved in oil phase solvent, and is protected from light 50
~60 DEG C of 10~12min of stirring, stir 1~1.5 hour at 20~25 DEG C, the emulsification oil phase component are made later.
Further, the preparation method includes：The dextrin modified casein polypeptide conjugate of the starch base is dissolved in phosphorus
In phthalate buffer, and 3h or more is stirred, albumen is made fully to be swollen, forms the aqueous phase solution.
The embodiment of the present invention additionally provides the dextrin modified casein polypeptide nanoemulsions of starch base prepared by preceding method.
Further, the morphosis of the dextrin modified casein polypeptide nanoemulsions of the starch base is spherical shape, grain size point
Cloth is 50~150nm.
The embodiment of the present invention additionally provides the dextrin modified casein polypeptide nanoemulsions of starch base above-mentioned in nutriment
Delivering, the controlled release of drug, the purposes in the stabilization field of bioactive substance.
Compared with prior art, advantages of the present invention includes：
The present invention is covalently connect using the preferable casein polypeptide of emulsion stability, starch base dextrin as raw material, using Mei Lade
The dextrin modified casein polypeptide conjugate of starch base is made as emulsifier in branch reaction, passes through high-pressure homogeneous and high pressure microjet skill
Art prepares the oil-in-water type nano-emulsion liquid and carrier system of the dextrin modified casein hydrolysis stabilizing polypeptides of certain degree of polymerization, junket
Albumen is modified by enzyme hydrolysis and dextrin covalence graft, and casein polypeptide solubility increases, and emulsibility and emulsion stability increase
By force, antioxygenic property improves, and antibacterial ability has a degree of improvement；Meanwhile nano oil-in-water emulsion tool prepared by the present invention
There are Nano Particle, small drop sizes to allow high-efficiency delivery, easily transhipment to absorb, hydrophobic bioactive material absorbing can be improved
Efficiency has the advantages that preferable antioxidant activity, antibiotic property, preferable emulsibility and emulsion stability, good bioencapsulation
Property and the effect of active material high-efficiency delivery, in the delivering of nutriment, the fields such as stabilization of the controlled release of drug, bioactive substance
With extensive use.
Specific implementation mode
In view of technical problem of the existing technology, inventor is able to propose this through studying for a long period of time and largely putting into practice
The technical solution of invention.The technical solution, its implementation process and principle etc. will be further explained as follows.
The present invention, through ultrasonic wave added enzyme hydrolysis at casein polypeptide, passes through Pullulanase mainly using casein as raw material
De- branch waxy corn starch combination polyethylene glycol sedimentation sizing method prepares the starch base dextrin of certain degree of polymerization, by casein polypeptide and
Starch base dextrin carries out covalence graft reaction, using high-pressure homogeneous and microjet method, prepares the dextrin modified junket of starch base
Polypeptide oil-in-water type nano-emulsion liquid.
A kind of preparation for the dextrin modified casein polypeptide conjugate of starch base that the one side of the embodiment of the present invention provides
Method comprising：
Casein polypeptide and starch base dextrin are provided；
Make the mixed system comprising the casein polypeptide and starch base dextrin carry out Mei Lade in 80~120 DEG C covalently to connect
Branch 90~120min of reaction, forms the dextrin modified casein polypeptide conjugate of starch base.
In some embodiments, the mass ratio of the casein polypeptide and starch base dextrin is 1：1~2.
In some embodiments, the preparation method includes：The casein polypeptide is dissolved in phosphate buffer, it
After the starch base dextrin is added.
Further, the preparation method further includes：After the completion of the Mei Lade covalence grafts react, by the reaction
System cools down, and freeze-drying is to -20 DEG C or less later.
Among some more preferably case study on implementation, the preparation method specifically includes：
Casein polypeptide is dissolved in phosphate buffer, by 1：Starch base dextrin is added in 2 ratio, is sufficiently stirred molten
Solution.Gained mixture is heated 90 minutes to form maillard reaction product, then under room temperature (25 DEG C) in 100 DEG C of water-baths
It is cooling.Then mixture is lyophilized 24 hours in freeze drier, is then stored at -20 DEG C until further using.
Further, the preparation method includes：Under conditions of pH value is 6.8~7.5, handled using ultrasonic wave added,
So that casein and trypsase is carried out 90~120min of enzymatic hydrolysis reaction in 40~60 DEG C, obtains casein polypeptide.
Further, the degree of hydrolysis of the casein polypeptide is 10~20%.
Further, the casein includes the caseic aqueous solution that casein content is 5~15wt%.
Further, the additive amount of the trypsase is 50~60U/g.
Further, for the temperature that ultrasonic wave added processing uses for 60~70 DEG C, power is 450~500W, and the time is
40~60min.
Further, the preparation method further includes：After the completion of the enzymatic hydrolysis reaction, to the enzymatic hydrolysis reaction
Inhibitor inactivation trypsase is added in reaction solution and terminates reaction.
Further, the casein polypeptide has exposed amino acid side chain amino.
Among some more preferably case study on implementation, the preparation method of the casein polypeptide specifically may include：
Casein (10% aqueous solution) is reacted with the trypsase of the additive amount of 50U/g in 50 DEG C of water-baths, is adjusted
PH to 7.0 is saved, restricted hydrolysis is carried out to albumen using ultrasonic wave added (64 DEG C of temperature, power 460W, time 60min), in water
It detects pH in solution preocess to change and the NaOH of 1mol/L is used to adjust, addition inhibitor inactivation trypsase termination is anti-after reacting 2h
It answers.Obtain the casein polypeptide that degree of hydrolysis is 10~20%.
Wherein, degree of hydrolysis is that functional group exposes in 10~20% casein polypeptides, has better solubility, emulsibility
And emulsion stability.Amino acid side chain amino reacts shape with the hydroxyl of dextrin reduction end by Maillard in casein polypeptide
At amino covalent bond, solubility, inoxidizability, emulsibility and the emulsion stability of protein are improved after covalence graft.
In some embodiments, the preparation method includes：Waxy corn starch Heat Gelatinization is set to be cooled to 40~60 DEG C, it
Pullulanase is added afterwards and carries out de- branch processing 20~for 24 hours, obtains dextrin solution, then using polyethylene glycol gradually the precipitation method by dextrin
Solution obtains the uniform starch base dextrin of the degree of polymerization in 20~25 DEG C of water-baths 20~be classified for 24 hours.
Further, the degree of polymerization of the starch base dextrin is 70~100.
Further, the mass ratio of the Pullulanase (60U/g) and waxy corn starch is 1：20~25.
Among some more preferably case study on implementation, the preparation method of the starch base dextrin specifically may include：
Waxy corn starch is dispersed in acetate buffer (0.05M, pH5.0) and boils 30min, later in 121 DEG C of high pressures
1h sterilize to be gelatinized completely, is cooled to 50 DEG C, is hydrolyzed with ultrasonic wave added Pullulanase, the ethyl alcohol for being added excessive 95% stops hydrolysis
Reaction for 24 hours in 40 DEG C of oven dryings crushes, sieving.It is dissolved in distilled water again, the dextrin for configuring 1.0%-4.0% is molten
5g polyethylene glycol heating stirring is added to clarifying in liquid, every 100mL solution, and for 24 hours, appearance carries out after precipitating for water-bath after being cooled to 25 DEG C
Centrifugation, is repeated several times, and for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtains the starch base dextrin that the degree of polymerization is 70-100.Sediment
It is washed with ethyl alcohol after centrifugation, drying and grinding into powder.
The other side of the embodiment of the present invention additionally provides the dextrin modified casein of starch base prepared by preceding method
Polypeptide conjugate.
The other side of the embodiment of the present invention additionally provides a kind of dextrin modified casein polypeptide nanoemulsions of starch base
Preparation method comprising：
The dextrin modified casein polypeptide conjugate of starch base is prepared according to preceding method；
Using high-pressure homogeneous and micro jetting technology, keep the water phase of the dextrin modified casein polypeptide conjugate of the starch base molten
Liquid is uniformly mixed with emulsification oil phase component, obtains the dextrin modified casein polypeptide nanoemulsions of starch base of oil-in-water type.
In some embodiments, the preparation method specifically includes：Using high pressure homogenization technique, to the emulsification oil phase group
Divide and the mixed liquor of aqueous phase solution carries out 2~5min of shearing in 15 DEG C or less, forms thick lotion, use high pressure microjet skill later
The thick lotion is further homogenized processing by art at 100~120MPa, obtains the dextrin modified casein polypeptide of starch base
Nanoemulsions store at 2~6 DEG C later.
Further, the mass ratio of the emulsification oil phase component liquid compatible with water is 1：8~10.
Among some more preferably case study on implementation, the system of the dextrin modified casein polypeptide nanoemulsions of starch base
Preparation Method specifically may include：
Emulsification oil phase component liquid compatible with water presses 1：8~10 mass ratio shears 2~5min by high shear machine
Thick lotion is formed, it is then in the case where 100~120MPa is preferably 103.4MPa that thick lotion is further equal by high pressure microjet instrument
Prime number obtains the nanoemulsions that grain size is 50~150nm, keeps temperature to be less than 15 DEG C using trash ice during homogenizing, sample storage
It is stored under the conditions of 2~6 DEG C.
Further, the preparation method includes：So that hydrophobic bioactive substance is dissolved in oil phase solvent, and is protected from light 50
~60 DEG C of magnetic force 10~12min of stirring at low speed stir 1~1.5 hour to ensure to be completely dissolved at 20~25 DEG C, are made later
The emulsification oil phase component.
Further, the oil phase solvent includes corn oil, but not limited to this.
Further, the preparation method includes：By the dextrin modified casein polypeptide conjugate of the starch base (2.0%,
W/w it) is dissolved in phosphate buffer (pH7.0,10mmol/L), and magnetic agitation 3h or more, is sufficiently stirred, keep albumen fully molten
It is swollen, form the aqueous phase solution.
Wherein, one of one more preferably case study on implementation as the present invention, the preparation can also include specifically following
Step：
1, the preparation of casein polypeptide：By casein (10% aqueous solution) with the trypsase of the additive amount of 50U/g 50
It is reacted in DEG C water-bath, pH to 7.0 is adjusted, using ultrasonic wave added (64 DEG C of temperature, power 460W, time 60min) to albumen
Restricted hydrolysis is carried out, pH is detected in hydrolytic process and changes and the NaOH of 1mol/L is used to adjust, inhibitor is added after reacting 2h
It inactivates trypsase and terminates reaction.Obtain the casein polypeptide that degree of hydrolysis is 10%~20%.
2, the preparation of starch base dextrin：Waxy corn starch is dispersed in acetate buffer (0.05M, pH5.0) and is boiled
30min is cooled to 50 DEG C, is hydrolyzed with Pullulanase in 121 DEG C of high pressure sterilization 1h to be gelatinized completely later, is added excessive 95%
Ethyl alcohol stop hydrolysis, for 24 hours in 40 DEG C of oven dryings, crush, sieving.It is dissolved in distilled water again, configures 1.0%-
4.0% dextrin solution, 5g polyethylene glycol heating stirring is added per 100mL solution to clarifying, after being cooled to 25 DEG C water-bath for 24 hours,
It is centrifuged after there is precipitation, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 70~100.
3, the preparation of the dextrin modified casein polypeptide conjugate of starch base：A certain amount of casein peptide is dissolved in phosphate to delay
In fliud flushing, starch base dextrin is added by a certain percentage, heating water bath 90min forms Maillard reaction conjugate, room at 100 DEG C
The lower cooling of temperature, low-temperature storage after freeze-drying.
4, emulsification oil phase component is prepared：Hydrophobic bioactive substance is dissolved in corn oil solvent, 60 DEG C of magnetic force are protected from light
Stirring at low speed 10min is then stirred at room temperature 1 hour to ensure to be completely dissolved, emulsification oil phase component is made.
5, aqueous phase solution is prepared：The dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base is dissolved in phosphate
In buffer solution (pH7.0,10mmol/L), magnetic agitation 3h or more is sufficiently stirred, and albumen is made fully to be swollen, and forms aqueous phase solution.
6, the preparation of oil-in-water type nano-emulsion liquid：It emulsifies oil phase component liquid compatible with water and presses 1：9 weight ratio passes through high pressure
Cutter shearing 2min forms thick lotion, then by high pressure microjet instrument at 103.4MPa by the further equal prime number of thick lotion
It is secondary to obtain the nanoemulsions that grain size is 50~150nm, keep temperature to be less than 15 DEG C using trash ice during homogenizing, sample is stored in
Under the conditions of 2~6 DEG C.
The other side of the embodiment of the present invention additionally provides the dextrin modified casein of starch base prepared by preceding method
Polypeptide nano lotion.
Further, the morphosis of the dextrin modified casein polypeptide nanoemulsions of the starch base is spherical shape, grain size point
Cloth is 50~150nm.Gained nanoemulsions have preferable antioxidant activity, antibiotic property, emulsibility and emulsion stability, good
Bioencapsulation and the effect of active material high-efficiency delivery.
The other side of the embodiment of the present invention additionally provides the dextrin modified casein polypeptide nano-emulsion of starch base above-mentioned
Liquid in the delivering of nutriment, the controlled release of drug, bioactive substance stabilization field in purposes.
The mechanism that the present invention prepares the dextrin modified casein polypeptide nanoemulsions of starch base may is that：The present invention is to emulsify
The preferable casein polypeptide of stability, starch base dextrin are raw material, are reacted using Mei Lade covalence grafts and starch base dextrin is made
Modified casein polypeptide conjugate prepares certain degree of polymerization as emulsifier by high-pressure homogeneous and high pressure micro jetting technology
The oil-in-water type nano-emulsion liquid and carrier system, casein of dextrin modified casein hydrolysis stabilizing polypeptides pass through enzyme hydrolysis and dextrin
Covalence graft is modified, and casein polypeptide solubility increases, and emulsibility and emulsion stability enhancing, antioxygenic property improve, and resist
Bacterium ability has a degree of improvement.
There is nano oil-in-water emulsion prepared by the present invention Nano Particle, small drop sizes to allow high-efficiency delivery, easily
Transhipment absorbs, and hydrophobic bioactive material absorbing efficiency can be improved, and has preferable antioxidant activity, antibiotic property, preferable breast
The advantages that property changed and emulsion stability, good bioencapsulation and the effect of active material high-efficiency delivery, in passing for nutriment
It send, the fields such as the stabilization of the controlled release of drug, bioactive substance have extensive use.
Below by way of several embodiments technical solution that simultaneously present invention be described in more detail.However, selected embodiment
It is merely to illustrate the present invention, and is not limited the scope of the invention.
Embodiment 1
Weigh 10g caseins, dissolved with phosphate buffer, adjust pH to 7.5, concentration of substrate 5%, in test tube with
Trypsase (13000-20000 BAEE units mg is added in the additive amount of 50U/g-1Albumen), it is placed in 55 DEG C of water-baths and carries out enzyme
Solution reaction, while (64 DEG C of temperature, power 460W, time 50min) hydrolysis is assisted using ultrasonic wave, it is examined in hydrolytic process
It surveys pH to change and the NaOH of 1mol/L is used to adjust, reaction time 2h.After reaction, addition trypsin inhibitor inactivates pancreas
Protease obtains the casein polypeptide that degree of hydrolysis is 10%, is lyophilized preservation.Waxy corn starch is scattered in acetate buffer
It boils in (0.05M, pH5.0) and in the water of boiling 30 minutes, in 121 DEG C of high pressure sterilizations 1 hour to be gelatinized completely, is cooled to
50 DEG C, and press starch：Enzyme is 1：20 ratio is added Pullulanase (60U/g) and hydrolyzes 24 hours, and excessive 95% ethyl alcohol is added
Stop hydrolysis, sediment is collected by centrifuging 10 minutes 3500, washed three times with 95% ethyl alcohol, in 40 DEG C of baking oven
Middle drying 24 hours, is then ground into fine powder in mortar, sieving.It is dissolved in distilled water again, configures 1.0%-4.0%
Dextrin solution, 5g polyethylene glycol heating stirring is added per 100mL solution to clarifying, after being cooled to 25 DEG C water-bath for 24 hours, sink
It is centrifuged behind shallow lake, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 70-95.
Starch base dextrin obtained above is carried out covalence graft with casein polypeptide to react.It is molten to weigh 2g casein polypeptides
In the phosphate buffer of 1/15mol/L, by 1：Starch base dextrin is added in 2 ratio, is sufficiently stirred dissolving.Gained is mixed
Object heats 90 minutes to form maillard reaction product in 100 DEG C of water-baths, is then cooled down under room temperature (25 DEG C).Then it will mix
It closes object to be lyophilized in freeze drier 24 hours, is then stored at -20 DEG C until further using.
By the dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base be dissolved in phosphate buffer (pH7.0,
In 10mmol/L), magnetic agitation 3h or more makes albumen fully be swollen, and forms aqueous phase solution；A certain amount of beta carotene is molten
In corn oil, a concentration of 0.1% (w/w), magnetic stirrer, which is stirred well to, to be completely dissolved, and oil phase component is made.By oil phase
Component liquid compatible with water presses 1：9 weight ratio homogeneous 2min forms thick lotion, then by high pressure microjet instrument in 103.4MPa
It is lower by the further homogeneous of thick lotion 6 times, keep temperature to be less than 15 DEG C using trash ice during homogenizing.Gained nanoemulsions grain size is
120-150nm has better emulsibility and storage stability, carries simultaneously compared with the nanoemulsions only stablized by casein
High antioxidant activity and antibiotic property, are a kind of good bioactive substance carriers of functional characteristic.
Embodiment 2
Weigh 10g caseins, dissolved with phosphate buffer, adjust pH to 7.5, concentration of substrate 5%, in test tube with
Trypsase (13000-20000 BAEE units mg is added in the additive amount of 50U/g-1Albumen), it is placed in 55 DEG C of water-baths and carries out enzyme
Solution reaction, while (64 DEG C of temperature, power 460W, time 60min) hydrolysis is assisted using ultrasonic wave, it is examined in hydrolytic process
It surveys pH to change and the NaOH of 1mol/L is used to adjust, reaction time 100min.After reaction, addition trypsin inhibitor goes out
Trypsase living obtains the casein polypeptide that degree of hydrolysis is 10%, is lyophilized preservation.Waxy corn starch is scattered in acetate buffer
It is boiled in liquid (0.05M, pH5.0) and in the water of boiling 30 minutes, it is cooling in 121 DEG C of high pressure sterilizations 1 hour to be gelatinized completely
To 50 DEG C, and press starch：Enzyme is 1：20 ratio is added Pullulanase (60U/g) and hydrolyzes 24 hours, and excessive 95% second is added
Alcohol stops hydrolysis, and sediment is collected by centrifuging 10 minutes 3500, washed three times with 95% ethyl alcohol, in 40 DEG C of baking
It is 24 hours dry in case, it is then ground into fine powder in mortar, is sieved.It is dissolved in distilled water again, configures 1.0%-
4.0% dextrin solution, 5g polyethylene glycol heating stirring is added per 100mL solution to clarifying, after being cooled to 25 DEG C water-bath for 24 hours,
It is centrifuged after there is precipitation, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 70-95.
Starch base dextrin obtained above is carried out covalence graft with casein polypeptide to react.Weigh the casein polypeptide of 2g
It is dissolved in the phosphate buffer of 1/15mol/L, by 1：Starch base dextrin is added in 2 ratio, is sufficiently stirred dissolving.Gained is mixed
It closes object and heats 100 minutes in 100 DEG C of water-baths to form maillard reaction product, then cooled down under room temperature (25 DEG C).Then
Mixture is lyophilized 24 hours in freeze drier, is then stored at -20 DEG C until further using.
By the dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base be dissolved in phosphate buffer (pH7.0,
In 10mmol/L), magnetic agitation 3h or more makes albumen fully be swollen, and forms aqueous phase solution；A certain amount of beta carotene is molten
In corn oil, a concentration of 0.1% (w/w), magnetic stirrer, which is stirred well to, to be completely dissolved, and oil phase component is made.By oil phase
Component liquid compatible with water presses 1：9 weight ratio homogeneous 2min forms thick lotion, then by high pressure microjet instrument in 103.4MPa
It is lower by the further homogeneous of thick lotion 8 times, keep temperature to be less than 15 DEG C using trash ice during homogenizing.Gained nanoemulsions grain size is
50-130nm has better emulsibility and storage stability, improves simultaneously compared with the nanoemulsions only stablized by casein
Antioxidant activity and antibiotic property, are a kind of good bioactive substance carriers of functional characteristic.
Embodiment 3
Weigh 10g caseins, dissolved with phosphate buffer, adjust pH to 7.0, concentration of substrate 5%, in test tube with
Trypsase (13000-20000 BAEE units mg is added in the additive amount of 60U/g-1Albumen), it is placed in 60 DEG C of water-baths and carries out enzyme
Solution reaction, while (temperature 60 C, power 450W, time 60min) hydrolysis is assisted using ultrasonic wave, it is examined in hydrolytic process
It surveys pH to change and the NaOH of 1mol/L is used to adjust, reaction time 2h.After reaction, addition trypsin inhibitor inactivates pancreas
Protease obtains the casein polypeptide that degree of hydrolysis is 10%, is lyophilized preservation.Waxy corn starch is scattered in acetate buffer
It boils in (0.05M, pH5.0) and in the water of boiling 30 minutes, in 121 DEG C of high pressure sterilizations 1 hour to be gelatinized completely, is cooled to
40 DEG C, and press starch:Enzyme is 1：25 ratio is added Pullulanase (60U/g) and hydrolyzes 20 hours, and excessive 95% ethyl alcohol is added
Stop hydrolysis, sediment is collected by centrifuging 10 minutes 3500, washed three times with 95% ethyl alcohol, in 40 DEG C of baking oven
Middle drying 24 hours, is then ground into fine powder in mortar, sieving.It is dissolved in distilled water again, configures 1.0%-4.0%
Dextrin solution, 5g polyethylene glycol heating stirring is added per 100mL solution to clarifying, after being cooled to 25 DEG C water-bath for 24 hours, sink
It is centrifuged behind shallow lake, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 80-100.
Starch base dextrin obtained above is carried out covalence graft with casein polypeptide to react.It is molten to weigh 2g casein polypeptides
In the phosphate buffer of 1/15mol/L, by 1：Starch base dextrin is added in 1 ratio, is sufficiently stirred dissolving.Gained is mixed
Object heats 90 minutes to form maillard reaction product in 120 DEG C of water-baths, is then cooled down under room temperature (25 DEG C).Then it will mix
It closes object to be lyophilized in freeze drier 24 hours, is then stored at -20 DEG C until further using.
By the dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base be dissolved in phosphate buffer (pH7.0,
In 10mmol/L), magnetic agitation 3h or more makes albumen fully be swollen, and forms aqueous phase solution；A certain amount of beta carotene is molten
In corn oil, a concentration of 0.1% (w/w), magnetic stirrer, which is stirred well to, to be completely dissolved, and oil phase component is made.By oil phase
Component liquid compatible with water presses 1：10 weight ratio homogeneous 4min forms thick lotion, then by high pressure microjet instrument at 100MPa
By the further homogeneous of thick lotion 5 times, temperature is kept to be less than 15 DEG C using trash ice during homogenizing.Gained nanoemulsions grain size is
100-150nm has better emulsibility and storage stability, carries simultaneously compared with the nanoemulsions only stablized by casein
High antioxidant activity and antibiotic property, are a kind of good bioactive substance carriers of functional characteristic.
Embodiment 4
Weigh 10g caseins, dissolved with phosphate buffer, adjust pH to 7.5, concentration of substrate 5%, in test tube with
Trypsase (13000-20000 BAEE units mg is added in the additive amount of 50U/g-1Albumen), it is placed in 50 DEG C of water-baths and carries out enzyme
Solution reaction, while (68 DEG C of temperature, power 500W, time 40min) hydrolysis is assisted using ultrasonic wave, it is examined in hydrolytic process
It surveys pH to change and the NaOH of 1mol/L is used to adjust, reaction time 90min.After reaction, addition trypsin inhibitor goes out
Trypsase living obtains the casein polypeptide that degree of hydrolysis is 16%, is lyophilized preservation.Waxy corn starch is scattered in acetate buffer
It is boiled in liquid (0.05M, pH5.0) and in the water of boiling 30 minutes, it is cooling in 121 DEG C of high pressure sterilizations 1 hour to be gelatinized completely
To 50 DEG C, and press starch:Enzyme is 1：20 ratio is added Pullulanase (60U/g) and hydrolyzes 24 hours, and excessive 95% second is added
Alcohol stops hydrolysis, and sediment is collected by centrifuging 10 minutes 3500, washed three times with 95% ethyl alcohol, in 40 DEG C of baking
It is 24 hours dry in case, it is then ground into fine powder in mortar, is sieved.It is dissolved in distilled water again, configures 1.0%-
4.0% dextrin solution is added 5g polyethylene glycol heating stirring per 100mL solution and extremely clarifies, water-bath 22h after being cooled to 20 DEG C,
It is centrifuged after there is precipitation, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 80-100.
Starch base dextrin obtained above is carried out covalence graft with casein polypeptide to react.Weigh the casein polypeptide of 2g
It is dissolved in the phosphate buffer of 1/15mol/L, by 1：Starch base dextrin is added in 2 ratio, is sufficiently stirred dissolving.Gained is mixed
It closes object and heats 100 minutes in 100 DEG C of water-baths to form maillard reaction product, then cooled down under room temperature (25 DEG C).Then
Mixture is lyophilized 24 hours in freeze drier, is then stored at -20 DEG C until further using.
By the dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base be dissolved in phosphate buffer (pH7.0,
In 10mmol/L), magnetic agitation 3h or more makes albumen fully be swollen, and forms aqueous phase solution；A certain amount of beta carotene is molten
In corn oil, a concentration of 0.1% (w/w), magnetic stirrer, which is stirred well to, to be completely dissolved, and oil phase component is made.By oil phase
Component liquid compatible with water presses 1：9 weight ratio homogeneous 4min forms thick lotion, then by high pressure microjet instrument at 120MPa
By the further homogeneous of thick lotion 4 times, temperature is kept to be less than 15 DEG C using trash ice during homogenizing.Gained nanoemulsions grain size is
60-135nm has better emulsibility and storage stability, improves simultaneously compared with the nanoemulsions only stablized by casein
Antioxidant activity and antibiotic property, are a kind of good bioactive substance carriers of functional characteristic.
Embodiment 5
Weigh 10g caseins, dissolved with phosphate buffer, adjust pH to 6.8, concentration of substrate 5%, in test tube with
Trypsase (13000-20000 BAEE units mg is added in the additive amount of 55U/g-1Albumen), it is placed in 40 DEG C of water-baths and carries out enzyme
Solution reaction, while (temperature 70 C, power 460W, time 50min) hydrolysis is assisted using ultrasonic wave, it is examined in hydrolytic process
It surveys pH to change and the NaOH of 1mol/L is used to adjust, reaction time 2h.After reaction, addition trypsin inhibitor inactivates pancreas
Protease obtains the casein polypeptide that degree of hydrolysis is 20%, is lyophilized preservation.Waxy corn starch is scattered in acetate buffer
It boils in (0.05M, pH5.0) and in the water of boiling 30 minutes, in 121 DEG C of high pressure sterilizations 1 hour to be gelatinized completely, is cooled to
60 DEG C, and press starch:Enzyme is 1：25 ratio is added Pullulanase (60U/g) and hydrolyzes 22 hours, and excessive 95% ethyl alcohol is added
Stop hydrolysis, sediment is collected by centrifuging 10 minutes 3500, washed three times with 95% ethyl alcohol, in 40 DEG C of baking oven
Middle drying 24 hours, is then ground into fine powder in mortar, sieving.It is dissolved in distilled water again, configures 1.0%-4.0%
Dextrin solution, 5g polyethylene glycol heating stirring is added per 100mL solution to clarifying, water-bath 20h, sinks after being cooled to 22 DEG C
It is centrifuged behind shallow lake, for 24 hours then at 40 DEG C of dryings, crushes, sieving, obtain the starch base dextrin that the degree of polymerization is 75-100.
Starch base dextrin obtained above is carried out covalence graft with casein polypeptide to react.Weigh the casein polypeptide of 2g
It is dissolved in the phosphate buffer of 1/15mol/L, by 1：Starch base dextrin is added in 1.5 ratio, is sufficiently stirred dissolving.By gained
Mixture heats 120 minutes to form maillard reaction product in 80 DEG C of water-baths, is then cooled down under room temperature (25 DEG C).Then
Mixture is lyophilized 24 hours in freeze drier, is then stored at -20 DEG C until further using.
By the dextrin modified casein polypeptide conjugate (2.0%, w/w) of starch base be dissolved in phosphate buffer (pH7.0,
In 10mmol/L), magnetic agitation 3h or more makes albumen fully be swollen, and forms aqueous phase solution；A certain amount of beta carotene is molten
In corn oil, a concentration of 0.1% (w/w), magnetic stirrer, which is stirred well to, to be completely dissolved, and oil phase component is made.By oil phase
Component liquid compatible with water presses 1：8 weight ratio homogeneous 5min forms thick lotion, then by high pressure microjet instrument at 120MPa
By the further homogeneous of thick lotion 4 times, temperature is kept to be less than 15 DEG C using trash ice during homogenizing.Gained nanoemulsions grain size is
60-120nm has better emulsibility and storage stability, improves simultaneously compared with the nanoemulsions only stablized by casein
Antioxidant activity and antibiotic property, are a kind of good bioactive substance carriers of functional characteristic.
In conclusion the small drop sizes permission of the nanoemulsions obtained by the above-mentioned technical proposal of the present invention is efficiently passed
Send, hydrophobic bioactive material absorbing efficiency can be improved, have preferable antioxidant activity, antibiotic property, preferable emulsibility and
The advantages that emulsion stability, has extensive in the fields such as stabilization of the delivering of nutriment, the controlled release of drug, bioactive substance
Purposes.
In addition, mode of the inventor referring also to embodiment 1-5, with the other raw materials and condition listed in this specification
Etc. being tested, and equally it has been made and has had preferable antioxidant activity, antibiotic property, preferable emulsibility and emulsion stability etc.
The dextrin modified casein polypeptide nanoemulsions of starch base of advantage.
It should be understood that the technical concepts and features of above-described embodiment only to illustrate the invention, ripe its object is to allow
The personage for knowing technique cans understand the content of the present invention and implement it accordingly, and the protection model of the present invention can not be limited with this
It encloses.Any equivalent change or modification in accordance with the spirit of the invention should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of the dextrin modified casein polypeptide conjugate of starch base, it is characterised in that including：
Casein polypeptide and starch base dextrin are provided；
Keep the mixed system comprising the casein polypeptide and starch base dextrin anti-in 80~120 DEG C of progress Mei Lade covalence grafts
90~120min is answered, the dextrin modified casein polypeptide conjugate of starch base is formed.
2. preparation method according to claim 1, it is characterised in that：The quality of the casein polypeptide and starch base dextrin
Than being 1：1~2；Preferably, the preparation method includes：The casein polypeptide is dissolved in phosphate buffer, Zhi Houjia
Enter the starch base dextrin；Preferably, the preparation method further includes：It, will after the completion of the Mei Lade covalence grafts react
The reaction system cooling, freeze-drying is to -20 DEG C or less later.
3. preparation method according to claim 1 or 2, it is characterised in that including：Under conditions of pH value is 6.8~7.5,
It is handled using ultrasonic wave added, casein and trypsase is made to carry out 90~120min of enzymatic hydrolysis reaction in 40~60 DEG C, obtain junket
Polypeptide；Preferably, the degree of hydrolysis of the casein polypeptide is 10~20%；Preferably, the casein includes casein
Content is the caseic aqueous solution of 5~15wt%；Preferably, the additive amount of the trypsase is 50~60U/g；Preferably,
For the temperature that the ultrasonic wave added processing uses for 60~70 DEG C, power is 450~500W, and the time is 40~60min；Preferably,
The preparation method further includes：After the completion of the enzymatic hydrolysis reaction, it is added and inhibits into the reaction solution of the enzymatic hydrolysis reaction
Agent inactivates trypsase and terminates reaction；Preferably, the casein polypeptide has exposed amino acid side chain amino.
4. preparation method according to claim 1 or 2, it is characterised in that including：Waxy corn starch Heat Gelatinization is set to cool down
To 40~60 DEG C, Pullulanase is added later and carries out de- branch processing 20~for 24 hours, obtain dextrin solution, then using polyethylene glycol by
Dextrin solution in 20~25 DEG C of water-baths 20~be classified for 24 hours, is obtained the uniform starch base dextrin of the degree of polymerization by the step precipitation method；It is excellent
Choosing, the degree of polymerization of the starch base dextrin is 70~100；Preferably, the mass ratio of the Pullulanase and waxy corn starch
It is 1：20~25.
5. the dextrin modified casein polypeptide conjugate of starch base prepared by any one of claim 1-4 the methods.
6. a kind of preparation method of the dextrin modified casein polypeptide nanoemulsions of starch base, it is characterised in that including：
The dextrin modified casein polypeptide conjugate of starch base is prepared according to any one of claim 1-4 the methods；
Using high-pressure homogeneous and micro jetting technology, make the aqueous phase solution of the dextrin modified casein polypeptide conjugate of the starch base with
Emulsification oil phase component uniformly mixes, and obtains the dextrin modified casein polypeptide nanoemulsions of starch base of oil-in-water type.
7. preparation method according to claim 6, it is characterised in that including：Using high pressure homogenization technique, to the emulsification
The mixed liquor of oil phase component and aqueous phase solution carries out 2~5min of shearing in 15 DEG C or less, forms thick lotion, uses high pressure micro- later
The thick lotion is further homogenized processing by fluidics at 100~120MPa, obtains the dextrin modified junket egg of starch base
White polypeptide nano lotion, stores at 2~6 DEG C later；Preferably, the mass ratio of the emulsification oil phase component liquid compatible with water is
1：8~10.
8. the preparation method described according to claim 6 or 7, it is characterised in that including：Hydrophobic bioactive substance is set to be dissolved in
In oil phase solvent, and 50~60 DEG C of 10~12min of stirring are protected from light, are stirred 1~1.5 hour at 20~25 DEG C later, institute is made
State emulsification oil phase component；Preferably, the oil phase solvent includes corn oil；
And/or the preparation method includes：The dextrin modified casein polypeptide conjugate of the starch base is dissolved in phosphate-buffered
In liquid, and 3h or more is stirred, albumen is made fully to be swollen, forms the aqueous phase solution.
9. the dextrin modified casein polypeptide nanoemulsions of starch base prepared by any one of claim 6-8 the methods；It is excellent
Choosing, the morphosis of the dextrin modified casein polypeptide nanoemulsions of starch base is spherical shape, particle diameter distribution is 50~
150nm。
10. dextrin modified casein polypeptide nanoemulsions of starch base described in claim 9 are in the delivering of nutriment, drug
Controlled release, bioactive substance stabilization field in purposes.
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