CN103014111B - Whey protein hydrolysate rich in arginine and preparation method thereof - Google Patents
Whey protein hydrolysate rich in arginine and preparation method thereof Download PDFInfo
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Abstract
The invention provides a whey protein hydrolysate rich in arginine and a preparation method of the whey protein hydrolysate. The preparation method comprises the following steps of: preparing a whey protein water solution and thermally treating the whey protein water solution to denaturalize the protein; adding protease into the whey protein water solution for hydrolysis after the protein is denaturalized, wherein papain is firstly added for hydrolysis till the degree of hydrolysis is 35-45%, then the aminopeptidase is added for hydrolysis till the degree of hydrolysis is 55-65%, and then the hydrolysis is ended; and ultrasonically filtering and concentrating the solution via an ultrafiltration membrane after the hydrolysis is ended to obtain the whey protein hydrolysate rich in arginine. The whey protein hydrolysate is a soluble hydrolysate rich in arginine. As the arginine in the hydrolysate is mainly in the form of oligopeptides, the whey protein hydrolysate provided by the invention has the advantages of low allergen content, easiness in dissolving in aquatic products and high protein content.
Description
Technical field
The invention relates to a kind of arginic lactalbumin hydrolysate and preparation method thereof that is rich in.
Background technology
Arginine has very important effect at medicine and field of food.Arginine plays very important effect in the processes such as human hormone's secretion, cell fission, wound recovery.General patient will supplement the arginine of the high purity pharmaceutical grade of certain content at perioperatively, is conducive to so the fast quick-recovery of patient's postoperative wound and physical function.
Arginine product is in the market mainly pharmaceutical grade arginine, and mostly its preparation method is by microorganism is carried out to genetic modification, and technique is prepared and obtained by fermentation.Mostly a lot of disclosed methods of prior art such as Chinese patent application 01121882.7,02126904.1 are to adopt this microbe fermentation method.Although but the higher cost of arginine purity that the arginic method of preparation pharmaceutical grade obtains is also very high, what this arginine can not be long-time and a large amount of as medicine simultaneously takes, and the highly purified monomer arginine of this pharmaceutical grade easily makes part patient produce anaphylaxis.
Chinese patent application 200910019665.6 discloses a kind of arginine and preparation technology thereof, and the main raw material wherein adopting is hair, and this raw material is difficult for obtaining, and arginine yield is very low.
Chinese patent application 201110325755.5 provides a kind of prozyme technology hydrolyzing lactoalbumin to prepare the technique of peptone, but when the selected two kinds of enzymes of this technique act on whey-protein because the relation of restriction enzyme site can not effectively be prepared and is rich in arginic hydrolysate.
Summary of the invention
The present invention is mainly the problem existing in arginine product and preparation method thereof in above-mentioned prior art, provide a kind of adopt prepared by prozyme technology hydrolyzing lactoalbumin be rich in arginic nutritional prod and preparation method thereof, arginine in product is mainly that the form with small peptide exists, can, for patient provides supplementing of nutrition in postoperative, reduce anaphylaxis simultaneously.
Therefore, one object of the present invention is to provide a kind of preparation method who is rich in arginic lactalbumin hydrolysate.
Another object of the present invention is to provide a kind of according to described method, prepare be rich in arginic lactalbumin hydrolysate.
For reaching above-mentioned purpose, on the one hand, the invention provides a kind of arginic lactalbumin hydrolysate preparation method that is rich in, this preparation method comprises the steps:
(1) the preparation whey-protein aqueous solution, and heat-treat and make protein denaturation;
(2) in the whey-protein aqueous solution after protein denaturation, adding proteolytic enzyme to be hydrolyzed, is wherein first to add papain hydrolysis to degree of hydrolysis to reach 35～45%, then adds aminopeptidase to be jointly hydrolyzed into degree of hydrolysis to reach 55%～65%, stop hydrolysis;
(3) solution after termination hydrolysis is through ultra-filtration membrane ultrafiltration, concentrated, is rich in arginic lactalbumin hydrolysate described in obtaining.
In the present invention, used whey albumen can be to produce by methods known in the art the whey-protein obtaining, as whey isolate protein (WPI) or whey protein concentrate (WPC) etc.
The present invention adopts combinative enzyme hydrolysis technique hydrolyzing lactoalbumin, contriver finds under study for action, papoid as restriction endonuclease acts preferentially on substrate, increase along with content of peptides in substrate, add again excision enzyme aminopeptidase, selected papoid and aminopeptidase acting in conjunction are in whey-protein and the insoluble polypeptide of part, difference due to enzymolysis site, in prepared hydrolysate, the arginic small peptide existence form overwhelming majority is solvable state, interest rate ultrafiltration membrance filter technology is isolated the hydrolysate of solubility again, can obtain like this solubility arginine hydrolysate that purity is higher.
According to specific embodiment of the invention scheme, in method of the present invention, preferably in the whey-protein aqueous solution of control step (1) preparation, whey-protein weight concentration is 2%～10% (w/w, press the cubage of total solid, except special mark and explanation, content described in the present invention and ratio are weight content and ratio), more preferably 5%～8%.
According to specific embodiment of the invention scheme, in method of the present invention, preferably control lipid content in the whey-protein aqueous solution of step (1) preparation and be less than 0.1%, more preferably below 0.09%.The whey-protein aqueous solution of preparing can optionally carry out suitable centrifugal degreasing so that wherein lipid content meet this concentration and limit.
According to specific embodiment of the invention scheme, in method of the present invention, the protein denaturation of the whey-protein aqueous solution process and can adopt under any heat treating method in field, in the present invention, preferably the whey-protein aqueous solution is incubated to 5～20min at 60 ℃～90 ℃ and makes protein denaturation.
According to specific embodiment of the invention scheme, in the present invention, described papoid and aminopeptidase be commercially available acquisition all, and each zymin meets relevant industries standard-required.Conventionally after determining used enzyme preparation, the pH value scope and the temperature that are applicable to enzymic hydrolysis can be known.Method of the present invention preferably also comprises as required: before adding proteolytic enzyme to be hydrolyzed in the whey-protein aqueous solution after protein denaturation, the pH value of the whey-protein aqueous solution after protein denaturation is adjusted to 6.0～9.0 process.Regulate the conditioning agent of pH value can adopt conventional conditioning agent in affiliated field, as NaOH solution, HCl solution etc.The consumption of concrete zymin and hydrolysis temperature, hydrolysis time are suitably determined and adjust the requirement of degree of hydrolysis in can be according to the present invention.In preferred version of the present invention, for improving arginic content and productive rate in product, the content of whey-protein in the whey-protein aqueous solution of take is benchmark, should control papoid consumption in step (2) is 1.5%～4% of whey-protein weight, and aminopeptidase consumption is 1%～3% of whey-protein weight; More preferably, total consumption of controlling papoid and aminopeptidase is 3%～6% of whey-protein weight, and the weight ratio of papoid and aminopeptidase is 1: 0.5～1: 1.More specifically, add papain hydrolysis condition to may be controlled to 50 ℃～65 ℃ hydrolysis 1～2 hour, adding the common hydrolysising condition of aminopeptidase to control is 50 ℃～65 ℃ hydrolysis 1～2 hour.
According to preferred specific embodiments of the present invention, in method of the present invention, to control and to add papain hydrolysis to add aminopeptidase to be jointly hydrolyzed to degree of hydrolysis when 40% left and right, while being jointly hydrolyzed into degree of hydrolysis 60% left and right, deactivation stops hydrolysis.In the present invention, the mensuration of described degree of hydrolysis is carried out according to the routine operation in affiliated field.The routine operation in field under the mode that deactivation stops being hydrolyzed also can adopt preferably maintains 5～20min by hydrating solution at 85 ℃～95 ℃ in the present invention and with inactivator, stops hydrolysis after degree of hydrolysis reaches requirement.
According to preferred specific embodiments of the present invention, in method of the present invention, described in step (3), ultrafiltration condition is: ultra-filtration membrane molecular weight aperture 5000～10000Da, enters film operating pressure 0.2～1.5MPa, membrane operating pressure 0.05～0.20MPa.In addition, the working temperature of described ultra-filtration membrane is 30 ℃～50 ℃.
Hydrolyzed solution after ultrafiltration can be further concentrated to obtain the lactalbumin hydrolysate goods of different size according to demand.Described concentrated can be with reference to any feasible concentration operation in prior art, such as reverse osmosis membrane concentration etc.Conventionally be concentrated into solid content between 37%～42% scope, can be used as lactalbumin hydrolysate goods and sell.
According to specific embodiment of the invention scheme, can also the hydrolyzate after concentrated is further dry to obtain the lactalbumin hydrolysate goods of dry powder form.One, more specifically in embodiment, method of the present invention comprises the further lyophilize of the hydrolyzed solution after ultrafiltration and concentration to prepare the process of lyophilized powder.Described freezing dry process can carry out according to the routine operation in affiliated field.
By taking method for hydrolysis of the present invention, the hydrolysate solubility obtaining is good, and the total solids content of this hydrolyzate of take is benchmark, and wherein arginic content can reach more than 30%.And, in the present invention, being rich in arginic product yield high, can reach more than 45%.
In the present invention, yield method of calculation are:
The quality (w) * 100% of raw material before quality (the w)/reaction of lyophilized powder after yield=reaction.
On the other hand, the present invention also provides a kind of arginic lactalbumin hydrolysate that is rich in, and this hydrolyzate prepares according to the method for the invention.
According to specific embodiments, lactalbumin hydrolysate of the present invention can be the form of concentrated solution, can be also further dried dry powder form.The total solids content of this hydrolyzate of take is benchmark, the oligopeptides content that wherein molecular weight is 180～1000Da is more than 80% (conventionally between 82%～90%), the free aminoacid content that molecular weight is less than 180Da is 2%～4%, and lipid content is below 1%; Arginine content (comprising the arginine that is included in oligopeptides and free arginine) is more than 30%, can reach 32%～40% even higher.
In the present invention, the measurement of each data is all the ordinary methods according to affiliated field, and for example, protein peptide molecular weight distribution adopts high performance gel filtration chromatography to measure, and Contents of Amino Acids method is used commercially available automatic analyzer for amino acids.About allergen content, because anaphylactogen main in whey-protein is beta-lactoglobulin, in the present invention, be to measure allergen content by measuring beta-lactoglobulin, measuring method is the test kit method in conjunction with colour developing based on Ag-Ab.
In sum, the present invention adopts prozyme technology hydrolyzing lactoalbumin to prepare a kind of arginic lactalbumin hydrolysate that is rich in, process costs is lower, described lactalbumin hydrolysate is rich in arginine, because the arginine major part in hydrolysate exists with oligopeptides form, be easy to digest and assimilate, and allergen content is low, product is very easily water-soluble, is of high nutritive value.This product can contribute to the fast quick-recovery of postoperative patient at patient's long-term taking in postoperative for some time as nutritional supplementation product, for patient provides supplementing of nutrition in postoperative, greatly reduces patient's anaphylaxis simultaneously.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet that in the present invention's one specific embodiment, arginic lactalbumin hydrolysate is rich in preparation.
Embodiment
By specific embodiment, describe below the beneficial effect of implementation process of the present invention and generation in detail, be intended to help reader to understand better essence of the present invention and feature, not as restriction that can practical range to this case.
Embodiment 1:
Referring to Fig. 1, whey-protein water is mixed with to the solution of 8% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) be less than 0.09% to lipid content, solution after degreasing is incubated to 10 minutes in 90 ℃ of hot water baths so that protein denaturation, with the NaOH aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 7.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 55 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 2.6%, 1.5h is to degree of hydrolysis approximately 40% in hydrolysis, then add the Aminopeptidase P roteAX (Japanese Amano Enzyme Co., Ltd (Amano Enzyme)) that accounts for raw dairy white protein consumption 1.4% to be jointly hydrolyzed again about 2h to degree of hydrolysis 60% left and right, at the Water Unders of 90 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 6000Da, the film operating pressure of entering of ultra-filtration membrane is 1.5MPa, membrane operating pressure is 0.10MPa, the working temperature of ultra-filtration membrane is 30 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of the present embodiment again.
1, molecular weight distribution determination
Method: adopt high performance gel filtration chromatography to measure protein peptide molecular weight distribution in the hydrolyzate of the present embodiment, obtain the relative percentage of different molecular weight ranges peptide by areas of peak normalization method; And with GPC software data processing, obtain number-average molecular weight (Mn) and the weight-average molecular weight (Mw) of different molecular weight ranges peptide.
Result:
| Molecular weight ranges | Water thawing dry powder |
| ＞10000 | 0.44 |
| 10000～5000 | 1.95 |
| 5000～3000 | 2.52 |
| 3000～2000 | 3.69 |
| 2000～1000 | 5.18 |
| 1000～500 | 29.19 |
| 500～180 | 54.53 |
| ＜180 | 2.5 |
2, determined amino acid (use automatic analyzer for amino acids)
Amino acid composition is analyzed (g/100g)
| Amino acid | Content | Amino acid | Content |
| Aspartic acid (aspartic acid) | 3.970 | Methionine (methionine(Met)) | 1.257 |
| Threonine (Threonine) | 2.084 | Isoleucine (Isoleucine) | 2.794 |
| Serine (Serine) | 2.864 | Leueine (leucine) | 4.940 |
| Glutamic acid (L-glutamic acid) | 7.747 | Tyrosine (tyrosine) | 5.017 |
| Proline (proline(Pro)) | 3.835 | Phenylalanine (phenylalanine) | 2.012 |
| Glycine (glycine) | 1.216 | Histidine (Histidine) | 1.530 |
| Alanine (L-Ala) | 0.466 | Lysine (Methionin) | 2.992 |
| Cysteine (Gelucystine) | 2.981 | Arginine (arginine) | 33.498 |
| Valine (α-amino-isovaleric acid) | 0.154 |
3, allergen measurement method
In whey-protein, main anaphylactogen is beta-lactoglobulin, measuring method is test kit method: the principle of detection is antigen antibody reaction, micropore comprises beta-lactoglobulin, add standard substance, sample solution and beta-lactoglobulin antibody separately, the free emulative and beta-lactoglobulin antibody combination with fixing beta-lactoglobulin, after washing, add ELIAS secondary antibody, fully combination, washes off and there is no combined enzyme labelling thing.Add substrate and developer, breed, until there is the colourless blueness that changes into.Add stop bath.Color has blueness to become yellow, under 450nm, measures light absorption value.
Result: after measured, in the present embodiment, in the pulvis sample of gained lactalbumin hydrolysate, beta-lactoglobulin total content is 1.01mg/g.
In addition, after measured, in the present embodiment, hydrolysate yield is 55%, and the lipid content in hydrolysate is less than 1%.
Embodiment 2:
Whey-protein water is mixed with to the solution of 5% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) be less than 0.09% to lipid content, solution after degreasing is incubated to 15 minutes in 80 ℃ of hot water baths so that protein denaturation, with the HCl aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 6.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 60 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 1.8%, 1.0h is to degree of hydrolysis approximately 40% in hydrolysis, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 1.2% to be jointly hydrolyzed again about 1.5h to degree of hydrolysis 60% left and right, at the Water Unders of 90 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 5000Da, the film operating pressure of entering of ultra-filtration membrane is 0.4MPa, membrane operating pressure is 0.05MPa, the working temperature of ultra-filtration membrane is 45 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of the present embodiment again.
The oligopeptides content that after the present embodiment enzymolysis, hydrolysate molecular weight is 180～1000Da is 82.2%, and the free aminoacid content that molecular weight is less than 180Da is 3.87%; Arginic content is 36%, lipid content approximately 0.5%, and hydrolysate yield is 50%.In lyophilized powder, beta-lactoglobulin total content is 0.97mg/g.
Embodiment 3:
Whey-protein water is mixed with to the solution of 2% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) be less than 0.09% to lipid content, solution after degreasing is incubated to 15 minutes in 75 ℃ of hot water baths so that protein denaturation, with the NaOH aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 8.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 65 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 2.6%, be hydrolyzed about 2.0h to degree of hydrolysis approximately 38%, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 2.4% to be jointly hydrolyzed again about 1.0h to degree of hydrolysis 60% left and right, at the Water Unders of 85 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 10000Da, the film operating pressure of entering of ultra-filtration membrane is 1.0MPa, membrane operating pressure is 0.2MPa, the working temperature of ultra-filtration membrane is 50 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of the present embodiment again.
The oligopeptides content that after the present embodiment enzymolysis, hydrolysate molecular weight is 180～1000Da is 85.6%, and the free aminoacid content that molecular weight is less than 180Da is 3.45%; Arginic content is 37%, lipid content approximately 0.6%, and hydrolysate yield is 52%.In lyophilized powder, beta-lactoglobulin total content is 1.32mg/g.
Embodiment 4:
Whey-protein water is mixed with to the solution of 10% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) be less than 0.09% to lipid content, solution after degreasing is incubated to 20 minutes in 60 ℃ of hot water baths so that protein denaturation, with the NaOH aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 9.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 55 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 3%, be hydrolyzed about 1.5h to degree of hydrolysis approximately 40%, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 3% to be jointly hydrolyzed again about 1.5h to degree of hydrolysis 61% left and right, at the Water Unders of 95 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 8000Da, the film operating pressure of entering of ultra-filtration membrane is 1.5MPa, membrane operating pressure is 0.10MPa, the working temperature of ultra-filtration membrane is 35 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of the present embodiment again.
The oligopeptides content that after the present embodiment enzymolysis, hydrolysate molecular weight is 180～1000Da is 89.5%, and the free aminoacid content that molecular weight is less than 180Da is 2.11%; Arginic content is 38%, lipid content approximately 0.6%, and hydrolysate yield is 47%.In lyophilized powder, beta-lactoglobulin total content is 1.21mg/g.
Embodiment 5
Whey-protein water is mixed with to the solution of 6% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) to lipid content approximately 0.08%, solution after degreasing is incubated to 10 minutes in 70 ℃ of hot water baths so that protein denaturation, with the NaOH aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 7.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 65 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 1.58%, be hydrolyzed about 1.0h to degree of hydrolysis approximately 40%, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 1.42% to be jointly hydrolyzed again about 1.0h to degree of hydrolysis 60% left and right, at the Water Unders of 95 ℃, bathe deactivation and within 15 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 5000Da, the film operating pressure of entering of ultra-filtration membrane is 0.4MPa, membrane operating pressure is 0.05MPa, the working temperature of ultra-filtration membrane is 45 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of the present embodiment again.
The oligopeptides content that after the present embodiment enzymolysis, hydrolysate molecular weight is 180～1000Da is 80.2%, and the free aminoacid content that molecular weight is less than 180Da is 3.9%; Arginic content is 32%, lipid content approximately 0.6%, and hydrolysate yield is 44%.In lyophilized powder, beta-lactoglobulin total content is 0.94mg/g.
Comparative example 1
Whey-protein water is mixed with to the solution of 15% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) to lipid content approximately 0.09%, solution after degreasing is incubated to 10 minutes in 80 ℃ of hot water baths so that protein denaturation, with the NaOH aqueous solution of 1mol/L, the pH value of the solution after this degreasing is adjusted to 8.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 65 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 1.5%, be hydrolyzed about 0.5h to degree of hydrolysis approximately 35%, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 0.5% to be jointly hydrolyzed again about 1.0h to degree of hydrolysis 50% left and right, at the Water Unders of 85 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 3000Da, the film operating pressure of entering of ultra-filtration membrane is 1.5MPa, membrane operating pressure is 0.10MPa, the working temperature of ultra-filtration membrane is 30 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of this comparative example again.
The oligopeptides content that after this comparative example enzymolysis, hydrolysate molecular weight is 180～1000Da is 65.2%, and the free aminoacid content that molecular weight is less than 180Da is 1.95%; Arginic content is 14%, lipid content approximately 0.6%, and hydrolysate yield is 34%.In lyophilized powder, beta-lactoglobulin total content is 0.91mg/g.
Comparative example 2
Whey-protein water is mixed with to the solution of 1% weight, and by this solution centrifugal degreasing (5000r/min, 55 ℃) to lipid content approximately 0.08%, solution after degreasing is incubated to 10 minutes in 100 ℃ of hot water baths so that protein denaturation, with the aqueous hydrochloric acid of 1mol/L, the pH value of the solution after this degreasing is adjusted to 4.0 after afterwards solution being cooled to room temperature, then lactoalbumin soln is placed in to 70 ℃ of water bath heat preservations, in lactoalbumin soln, add the papoid preparation that accounts for raw dairy white protein consumption 0.33%, be hydrolyzed about 0.5h to degree of hydrolysis approximately 20%, then add the Aminopeptidase P roteAX that accounts for raw dairy white protein consumption 0.67% to be jointly hydrolyzed again about 0.5h to degree of hydrolysis 35% left and right, at the Water Unders of 85 ℃, bathe deactivation and within 10 minutes, stop hydrolysis, recycling ultra-filtration membrane is processed this hydrating solution membrane filtration, the membrane molecule metering-orifice footpath of ultra-filtration membrane is 12000Da, the film operating pressure of entering of ultra-filtration membrane is 1.5MPa, membrane operating pressure is 0.10MPa, the working temperature of ultra-filtration membrane is 30 ℃, permeate is utilized reverse osmosis membrane to carry out concentration, freeze-drying concentrated solution afterwards, makes the pulvis sample that is rich in arginic lactalbumin hydrolysate of this comparative example again.
The oligopeptides content that after this comparative example enzymolysis, hydrolysate molecular weight is 180～1000Da is 45.9%, and the free aminoacid content that molecular weight is less than 180Da is 2.12%; Arginic content is 10%, lipid content approximately 0.6%, and hydrolysate yield is 29%.In lyophilized powder, beta-lactoglobulin total content is 1.56mg/g.
Claims (7)
1. be rich in a preparation method for arginic lactalbumin hydrolysate, this preparation method comprises the steps:
(1) the preparation whey-protein aqueous solution, and heat-treat and make protein denaturation;
(2) in the whey-protein aqueous solution after protein denaturation, adding proteolytic enzyme to be hydrolyzed, is wherein first to add papain hydrolysis to degree of hydrolysis to reach 35～45%, then adds aminopeptidase to be jointly hydrolyzed into degree of hydrolysis to reach 55%～65%, stop hydrolysis; Wherein, the content of whey-protein in the whey-protein aqueous solution of take is benchmark, controlling papoid consumption in step (2) is 1.5%～4% of whey-protein weight, aminopeptidase consumption is 1%～3% of whey-protein weight, total consumption of controlling papoid and aminopeptidase is 3%～6% of whey-protein weight, and the weight ratio of papoid and aminopeptidase is 1:0.5～1:1; Wherein adding papain hydrolysis condition is 50 ℃～65 ℃ hydrolysis 1～2 hour, and adding the common hydrolysising condition of aminopeptidase is 50 ℃～65 ℃ hydrolysis 1～2 hour;
(3) solution after termination hydrolysis is through ultra-filtration membrane ultrafiltration, concentrated, is rich in arginic lactalbumin hydrolysate described in obtaining; Described ultrafiltration condition is: ultra-filtration membrane molecular weight aperture 5000～10000Da, enters film operating pressure 0.2～1.5MPa, membrane operating pressure 0.05～0.20MPa.
2. method according to claim 1, wherein, controlling whey-protein weight concentration in the whey-protein aqueous solution of step (1) preparation is 2%～10%, and controls lipid content in this whey-protein aqueous solution and be less than 0.1%; This whey-protein aqueous solution is at 60 ℃～90 ℃, be incubated 5～20min and make protein denaturation.
3. method according to claim 1, wherein, in step (2), degree of hydrolysis reaches and hydrating solution is maintained to 5～20min at 85 ℃～95 ℃ after requirement and with inactivator, stop hydrolysis.
4. method according to claim 1, the method also comprises: before adding proteolytic enzyme to be hydrolyzed in the whey-protein aqueous solution after protein denaturation, the pH value of the whey-protein aqueous solution after protein denaturation is adjusted to 6.0～9.0 process.
5. method according to claim 1, the method also comprises the further lyophilize of the hydrolyzed solution after ultrafiltration and concentration to prepare the process of lyophilized powder.
6. be rich in an arginic lactalbumin hydrolysate, this hydrolyzate prepares according to method described in claim 1～5 any one.
7. lactalbumin hydrolysate according to claim 6, the total solids content of this hydrolyzate of take is benchmark, the oligopeptides content that wherein molecular weight is 180～1000Da is more than 80%, and the free aminoacid content that molecular weight is less than 180Da is 2%～4%, and lipid content is below 1%; Arginine content is more than 30%.
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| CN201210538587.2A CN103014111B (en) | 2012-12-13 | 2012-12-13 | Whey protein hydrolysate rich in arginine and preparation method thereof |
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| CN201210538587.2A CN103014111B (en) | 2012-12-13 | 2012-12-13 | Whey protein hydrolysate rich in arginine and preparation method thereof |
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| CN103014111B true CN103014111B (en) | 2014-11-12 |
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| CN108410938A (en) * | 2018-04-24 | 2018-08-17 | 浙江大学 | A method of preparing low bitter taste lactalbumin antioxidant peptide powder |
| CN108611390A (en) * | 2018-04-24 | 2018-10-02 | 浙江大学 | A method of preparing low bitter taste buffalo's milk casein antioxidant peptide powder |
| CN114630584A (en) * | 2019-11-07 | 2022-06-14 | 纽崔西亚公司 | Deep whey protein hydrolysate with tolerogenic peptides |
| CN114190469B (en) * | 2021-12-27 | 2024-03-15 | 南通励成生物工程有限公司 | Hydrolyzed whey protein composite amino acid chelate as well as preparation method and application thereof |
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| JP5750039B2 (en) * | 2008-03-26 | 2015-07-15 | グランビア ニュートリショナルズ （アイルランド） リミテッド | Leucine-rich peptide composition and isolation method |
| CN102093469B (en) * | 2009-12-11 | 2014-07-30 | 光明乳业股份有限公司 | Milk-derived antioxidant active peptide and application thereof |
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| CN102628073B (en) * | 2012-03-28 | 2013-11-20 | 东北农业大学 | Glycosylated lactalbumin polypeptide with antioxidant activity as well as preparation method and application of glycosylated lactalbumin polypeptide |
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