CN108410938A - A method of preparing low bitter taste lactalbumin antioxidant peptide powder - Google Patents
A method of preparing low bitter taste lactalbumin antioxidant peptide powder Download PDFInfo
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- lactalbumin
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The invention discloses a kind of method preparing low bitter taste lactalbumin antioxidant peptide powder, this method includes：Aqueous whey protein solution is prepared, after high-temperature denatured, neutral proteinase is added and carries out primary enzymolysis, add ProteAX protease and carry out secondary enzymolysis, after enzymolysis, high temperature enzyme deactivation obtains lactalbumin enzymolysis liquid；By lactalbumin enzymolysis liquid successively through centrifuging with after hyperfiltration treatment, freeze-drying obtains lactalbumin antioxidant peptide powder.The present invention carries out two-stage enzymolysis using neutral proteinase and ProteAX protease to lactalbumin, obtains the lactalbumin antioxidant peptide powder that nitrogen recovery is high, molecular weight is small, antioxidant activity is strong and bitter taste is low.
Description
Technical field
The present invention relates to dairy products technical field more particularly to a kind of sides preparing low bitter taste lactalbumin antioxidant peptide powder
Method.
Background technology
Milk and milk products are one of food most long in human diet history, and the abundant protein contained by them is not only
Abundant nutrition can be provided, and be the important sources of biologically active peptide.Lactalbumin is detached after cow's milk precipitates casein
The general name of the multiple proteins component gone out accounted for newborn total protein content once as industrialized production cheese and the byproduct of casein
20% or so.
In recent years, it is increasingly paid close attention to by people about the peptide fragment with physiological activity contained in milk and milk products.It is small
Transhipment, absorption and utilization rate of the molecular peptide in enteron aisle are better than single amino acid, and by its different amino acid range side
Formula can have different physiological activity, such as participate in immunological regulation in vivo, remove free radical, adjust blood pressure balance, antithrombotic
With strengthen bone health etc..Wherein antioxidation active peptides are a studied wide directions, in a variety of breasts and breast
Different anti-oxidant segments is identified in product, is contained abundant hydrophobic amino acid and basic amino acid in lactalbumin, is
Prepare the quality raw materials of anti-oxidation peptide.
In recent years, it is increasingly paid close attention to by people about the peptide fragment with physiological activity contained in milk whey protein.
Transhipment, absorption and utilization rate of the small-molecular peptides in enteron aisle are better than single amino acid, and by its different amino acid range
Mode can have different physiological activity, such as participate in immunological regulation in vivo, remove free radical, adjust blood pressure balance, anti-blood
Bolt and reinforcing bone health etc..Wherein antioxidation active peptides are a studied wide directions, it is a variety of breast and
Different anti-oxidant segments is identified in dairy products, contains abundant hydrophobic amino acid and basic amino acid in lactalbumin,
It is the quality raw materials for preparing anti-oxidation peptide.
Enzymatic isolation method is the most popular method of current industrially prepared active peptide, and this method mild condition, controllability are strong, product is steady
It is fixed.But in enzymolysis process, hydrophobic amino acid from protein cleavage at polypeptide when expose, often make C-terminal have hydrophobicity ammonia
The polypeptide of base acid carries bitter taste, and the organoleptic quality of peptide product is caused to reduce.Addition adsorbent or embedding medium are often used in industry
Mode removes or so that bitter taste is reduced in conjunction with hydrophobic amino acid, but these modes can bring necessary amino acid loss, reduce cow's milk
The problems such as nutritive value of, raising finished product itself.Using endopeptidase and peptide ending enzyme (aminopeptidase and carboxypeptidase) synergetic hydrolysis
Mode to reduce polypeptide products bitter taste have positive effect.First with the endo protease that restriction enzyme site is hydrophobic amino acid
Scinderin matter, release end position carry the small-molecular peptides of hydrophobic amino acid, and dredging for bitter peptides C-terminal is cut off followed by aminopeptidase
Aqueous amino acid or the hydrophobic amino acid that bitter peptides N-terminal is cut off using carboxypeptidase achieve the effect that reduce bitter taste.But inside and outside
During peptase synergetic hydrolysis, the condition that different protease are suitble to is different, constantly regulate soda acid and temperature is needed, in practical life
It also needs to increase desalination process in production.
Therefore, it finds hydrolysis ability by force and the matched de- bitter protease of hydrolysis environment is most important for active peptide industry
's.
Invention content
The present invention provides a kind of methods preparing low bitter taste lactalbumin antioxidant peptide powder, are prepared using this method
Lactalbumin small peptide not only nitrogen recovery is high, molecular weight is low, and antioxidant activity is strong, to DPPH free radicals and ABTS free radicals
Clearance rate is higher, and can significantly reduce the bitter taste of product, is conducive to the popularization of product.
Specific technical solution is as follows：
A method of low bitter taste lactalbumin antioxidant peptide powder is prepared, including：
(1) aqueous whey protein solution is prepared, after high-temperature denatured, neutral proteinase is added and carries out primary enzymolysis, adds
ProteAX protease carries out secondary enzymolysis, and after enzymolysis, high temperature enzyme deactivation obtains lactalbumin enzymolysis liquid；
(2) by lactalbumin enzymolysis liquid successively through centrifuging with after hyperfiltration treatment, it is anti-oxidant to obtain lactalbumin for freeze-drying
Gly-His-Lys.
Neutral proteinase used in the present invention is purchased from Nanning Pang Bo bioengineering Co., Ltd, derives from withered grass gemma
Bacillus, neutral protease vigor 200,000U/g；ProteAX protease used is purchased from Japanese amano enzyme preparation company, is one
Kind aminopeptidase, exopeptidase vigor made of aspergillus oryzae (Aspergillus oryzae) fermentation refinement are 1400U/g.
Used whey albumen of the present invention is concentrated type PURE WHEY (WPC), and protein content of whey is more than 80%, purchased from new
Xi Lan dairy company Fonterras；WPC90, WPC75 or WPC35 can also be used in concrete operations.
Enzymolysis efficiency can be improved in suitable albumen concentration, increases nitrogen recovery, and albumen concentration is too low to increase manufacturing cost,
The too high protease that is unfavorable for of albumen concentration efficiently digests.Preferably, in the aqueous whey protein solution, the matter of lactalbumin
It is 2~8% to measure score.
To ensure that lactalbumin is substantially dissolved in water, lactalbumin need to be placed in 90 DEG C~100 DEG C of water and stir 10
~20min, the course of dissolution are also degenerative process simultaneously.Preferably, in step (1), the high-temperature denatured temperature is 90
~100 DEG C, the time is 10~20min.High temperature can destroy the stereochemical structure of protein, expose peptide chain, be conducive to protease enzyme
It cuts, hence it is evident that improve digesting efficiency.
Neutral proteinase is not necessarily to use acid-base accommodation pH value of solution suitable for the native pH of lactoalbumin soln.Protease adds
Dosage has an impact the molecular size range and antioxygenic property of nitrogen recovery, lactalbumin anti-oxidation peptide.Preferably, step
(1) in, the mass ratio of the neutral proteinase and lactalbumin is 1~3:100.
Different enzymatic hydrolysis conditions can obtain the enzymolysis liquid of different final state pH, influence follow-up ProteAX protease effect and play, because
This can get the enzymolysis liquid that the follow-up Prote AX protease of optimum takes off hard work sequence suitable for enzymatic hydrolysis condition.Preferably, step (1)
In, the temperature of the primary enzymolysis is 40~60 DEG C, and the time is 2~5h.The enzymatic hydrolysis condition can efficiently play proteinase activity,
Improve polypeptide yield.
The additive amount of ProteAX protease can have an impact the bitter taste of lactalbumin anti-oxidation peptide, meanwhile, Prote AX tools
Have certain endopeptidase activity, can further scinderin matter and polypeptide, influence nitrogen recovery, peptide molecular weight and anti-oxidant energy
Power.Preferably, in step (1), the mass ratio of the ProteAX protease and lactalbumin is 1~3:100.
Preferably, in step (1), the temperature of the secondary enzymolysis is 40~60 DEG C, and the time is 2~5h.The enzymolysis item
Part can efficiently play proteinase activity, improve polypeptide yield, reduce the bitter taste of polypeptide.
Preferably, in step (2), the rotating speed of the centrifugation is 6000~8000r/min, and the time is 10~20min；It can
With isolated polypeptide and insoluble protein as far as possible, polypeptide yield is improved.
Preferably, in step (2), the interception of the ultrafiltration is 6000~10000Da, and pressure is 0.4~0.6MPa.
Ultrafiltration can remove a small number of residual lipids and high molecular weight protein and polypeptide, can effectively remove impurity, obtain limpid pure peptide liquid,
Improve the antioxygenic property of product.
Compared with prior art, the invention has the advantages that：
(1) present invention carries out two-stage enzymolysis using neutral proteinase and ProteAX protease to lactalbumin, obtains nitrogen and returns
The lactalbumin antioxidant peptide powder that high income, molecular weight are small, antioxidant activity is strong and bitter taste is low.
(2) the method for the present invention only needs a step enzyme deactivation, not only green energy conservation without adjusting pH and temperature repeatedly, but also
Follow-up desalting steps are reduced, industrial continuous large-scale production is easy to.
(3) the enzyme hydrolysis ability that the method for the present invention uses is strong, and nitrogen recovery is high, and Gly-His-Lys molecular weight is small, and oxidation resistance is strong,
Sense organ is good, can be separately as functional food or as additive application in other food.
Specific implementation mode
With reference to specific embodiment, the invention will be further described, and what is be exemplified below is only the specific implementation of the present invention
Example, but protection scope of the present invention is not limited only to this.
DPPH free radical scavenging activities, ABTS free radical scavenging activities, nitrogen recovery involved in the following example and molecular weight
The assay method of distribution is as follows：
(1) measurement of DPPH free radical scavenging activities：
1. being separately added into 0.1mL pure water and 1.9mL DPPH ethanol solutions (1mmoL/L) in 5mL centrifuge tubes, shake
After even after 30min is reacted in dark place, with the light absorption value A under UV spectrophotometer measuring its 515nmc；
2. being separately added into 0.1mL candidate polypeptides liquid (2mg/mL) in 5mL centrifuge tubes and 1.9mL DPPH absolute ethyl alcohols being molten
Liquid (1mmoL/L), after shaking up after 30min is reacted in dark place, with the light absorption value A under UV spectrophotometer measuring its 515nmi；
3. DPPH clearance rates following formula result of calculation：
(2) measurement of ABTS free radical scavenging activities：
1. prepared by ABTS free radicals：It is a concentration of that the potassium persulfate solution of 440 a concentration of 140mmoL/L of μ L is added to 25mL
In the ABTS solution of 7mmoL/L, mixing, which is protected from light after 12h, to be settled to 100mL and obtains ABTS radical reaction liquid.
2. being separately added into 0.1mL pure water and 1.9mL ABTS reaction solutions (2mmoL/L) in 5mL centrifuge tubes, mixing is shaken
The light absorption value A under its 734nm of UV spectrophotometer measuring is used after even reaction 10min0；
3. being separately added into 0.1mL candidate polypeptides liquid (2mg/mL) and 1.9mL ABTS reaction solutions in 5mL centrifuge tubes, mix
Close the light absorption value A used after shaking up reaction 10min under its 734nm of UV spectrophotometer measuring1；
4. ABTS clearance rates following formula result of calculation：
(3) nitrogen recovery measures：Measure the nitrogen percentage composition point in lactalbumin and Gly-His-Lys respectively by Kjeldahl's method
It Wei not N1And N2。
Nitrogen recovery is calculate by the following formula：
(4) small molecular weight titanium assay：Using GE AKTA pure protein purification instrument, it is equipped with Superdex Peptide
10/300GL chromatographic columns, mobile phase are 30% (V/V) acetonitrile solution containing 0.1% (V/V) TFA, flow velocity 0.5mL/min, detection
Wavelength is 215nm.The polypeptide solution of 2mg/mL is prepared, 100 μ L of sample introduction after 0.22 μm of water system filter membrane are crossed.Molecular weight standards are selected
Aprotinin (6511Da), bacitracin (1422Da), reduced glutathione (614Da) and tyrosine (181Da).The molecule of Gly-His-Lys
It is small-molecular peptides that amount, which is distributed within the scope of 200~1500Da, and the ratio of small molecular weight titanium is determined using different molecular weight standard items.
(5) subjective appreciation：Subjective appreciation is carried out using the peptide liquid of Gly-His-Lys and 10mg/mL.Selecting 8 has subjective appreciation base
Assessment officer's (4 male 4 female, 22~25 years old age) of plinth, the bitter taste, color and luster and solubility of Gly-His-Lys and peptide liquid are carried out according to grade form
It judges, for wherein bitterness value using the tanning solution of various concentration as evaluation criteria, 50mmol/L tanning solutions are faint bitter taste,
100mmol/L tanning solutions are apparent bitter taste, and 200mmol/L tanning solutions are strong bitterness.Summarize 8 parts of scorings, items are respectively
It is averaged as product score after removing best result and minimum point.
1 subjective appreciation table of table
Embodiment 1
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 2:100, constant temperature digests 3.5h, not enzyme deactivation after primary enzymolysis, continues to add ProteAX protease,
The mass ratio of ProteAX protease and lactalbumin is 1.5:100, constant temperature digests 3.5h, is boiled for 95 DEG C after digesting twice
Enzyme deactivation 10min obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Embodiment 2
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 2% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 55 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 0.5:100, constant temperature digests 3h, not enzyme deactivation after primary enzymolysis, continues to add ProteAX protease,
The mass ratio of ProteAX protease and lactalbumin is 0.5:100, constant temperature digests 3h, boils and goes out for 95 DEG C after digesting twice
Enzyme 10min obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Embodiment 3
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 45 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 1.5:100, constant temperature digests 4h, not enzyme deactivation after primary enzymolysis, continues to add ProteAX protease,
The mass ratio of ProteAX protease and lactalbumin is 1:100, constant temperature digests 3h, boils enzyme deactivation for 95 DEG C after digesting twice
10min obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Embodiment 4
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 8% aqueous whey protein solution to take mass fraction, and 15min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 60 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 2.5:100, constant temperature digests 2.5h, not enzyme deactivation after primary enzymolysis, continues to add ProteAX protease,
The mass ratio of ProteAX protease and lactalbumin is 1.5:100, constant temperature digests 2.5h, is boiled for 95 DEG C after digesting twice
Enzyme deactivation 10min obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 1
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 2:100, constant temperature digests 3h, boils enzyme deactivation 10min for 95 DEG C after enzymolysis, obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 2
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and ProteAX protease, ProteAX protease and breast is added
Albuminised mass ratio is 2:100, constant temperature digests 3h, boils enzyme deactivation 10min for 95 DEG C after enzymolysis, obtains lactalbumin enzymolysis
Liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 3
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 4% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and neutral proteinase, neutral proteinase and lactalbumin is added
Mass ratio be 1.5:100, constant temperature digests 3h, not enzyme deactivation after primary enzymolysis, continues to add flavor protease, flavor albumen
The mass ratio of enzyme and lactalbumin is 1.5:100, constant temperature digests 3h, boils enzyme deactivation 10min for 95 DEG C after digesting twice, obtains newborn
Albumin enzymolysis liquid.Flavor protease used is purchased from Nanning Pang Bo bioengineering Co., Ltd, is sent out from aspergillus oryzae
Ferment contains endo protease and two kinds of activity of circumscribed peptase.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 4
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 4% aqueous whey protein solution to take mass fraction, after 90 DEG C are boiled pretreatment 10min,
50 DEG C are cooled to, then is placed in magnetic force constant temperature water bath apparatus, is added in the ratio for adding 0.015g protease per 1g lactalbumins
Neutral proteinase, constant temperature digest 3h, and not enzyme deactivation after primary enzymolysis is cooled to 40 DEG C, by per 1g lactalbumin additions
The ratio of 0.015g protease adds Peptidase R protease, and 40 DEG C of constant temperature is kept to digest 3h, after digesting twice, boils
Enzyme deactivation is boiled, lactalbumin enzymolysis liquid is obtained.Peptidase R protease used is purchased from Japanese amano enzyme preparation company, is an introduces a collection
The aminopeptidase made of Rhizopus oryzae (Rhizopus oryzae) fermentation is refined, exopeptidase vigor are 420U/g.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 5
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 4% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen is cooled to 37 DEG C, then is placed in magnetic force constant temperature water bath apparatus, in the ratio for adding 0.015g protease per 1g lactalbumins
Trypsase is added, constant temperature digests 3h, not enzyme deactivation after primary enzymolysis, continues to add 0.010g albumen by every 1g lactalbumins
The ratio of enzyme adds ProteAX protease, and 37 DEG C of constant temperature is kept to digest 3h, after digesting twice, boils enzyme deactivation, obtains whey egg
White enzymolysis liquid.Trypsase used be purchased from Nanning Pang Bo bioengineering Co., Ltd, the extraction purification from pig pancreas and obtain.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 6
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 4% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and alkali protease, alkali protease and lactalbumin is added
Mass ratio be 1.5:100, constant temperature digests 3h, not enzyme deactivation after primary enzymolysis, continues to add ProteAX protease,
The mass ratio of ProteAX protease and lactalbumin is 1.5:100, constant temperature digests 3h, boils and goes out for 95 DEG C after digesting twice
Enzyme 10min obtains lactalbumin enzymolysis liquid.Alkali protease used is purchased from Nanning Pang Bo bioengineering Co., Ltd, comes
It is obtained, prolease activity 800,000U/g derived from the Bacillus licheniformis2709 fermentation that bacterial protoplast mutagenic and breeding goes out.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 7
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, while neutral proteinase and ProteAX protease is added, in
Property protease and lactalbumin mass ratio be 1.5:The mass ratio of 100, ProteAX protease and lactalbumin is also 1.5:
100, constant temperature digests 4h, boils enzyme deactivation 10min for 95 DEG C after enzymolysis, obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
Comparative example 8
A method of low bitter taste lactalbumin antioxidant peptide powder being prepared, particular content is as follows：
(1) protease hydrolyzed：It is 5% aqueous whey protein solution to take mass fraction, and 10min dissolving wheys are stirred at 90 DEG C
Albumen when being cooled to 50 DEG C, is placed in magnetic force constant temperature enzyme reactor, and ProteAX protease, ProteAX protease and breast is added
Albuminised mass ratio is 1.5:100, constant temperature digests 3h, not enzyme deactivation after primary enzymolysis, continues to add neutral proteinase, in
Property protease and lactalbumin mass ratio be 1.5:100, constant temperature digests 3h, boils enzyme deactivation for 95 DEG C after digesting twice
10min obtains lactalbumin enzymolysis liquid.
(2) preparation of Gly-His-Lys：After enzymolysis liquid 7000r/min centrifugations 15min, collection supernatant is thick peptide liquid, by thick peptide liquid
The ultrafiltration membrane that interception is 10kDa is crossed, pressure 0.5MPa sloughs lipid, high molecular weight protein and polypeptide, then freeze-dried,
Obtain lactalbumin Gly-His-Lys.
The lactalbumin Gly-His-Lys product prepared to above-described embodiment 1~4 and comparative example 1~8 carries out nitrogen recovery, small
It molecule peptide content, the measurement of DPPH free radical scavenging activities and ABTS free radical scavenging activities and bitter taste, color and luster and deliquescent comments
Fixed, the results are shown in Table 2.
The Comparative result of table 2 different Examples 1 to 4 and reference examples 1~8
Claims (9)
1. a kind of method preparing low bitter taste lactalbumin antioxidant peptide powder, which is characterized in that including：
(1) aqueous whey protein solution is prepared, after high-temperature denatured, neutral proteinase is added and carries out primary enzymolysis, adds ProteAX
Protease carries out secondary enzymolysis, and after enzymolysis, high temperature enzyme deactivation obtains lactalbumin enzymolysis liquid；
(2) by lactalbumin enzymolysis liquid successively through centrifuging with after hyperfiltration treatment, freeze-drying obtains lactalbumin anti-oxidation peptide
Powder.
2. the method as described in claim 1, which is characterized in that in the aqueous whey protein solution, the quality of lactalbumin
Score is 2~8%.
3. the method as described in claim 1, which is characterized in that in step (1), the matter of the neutral proteinase and lactalbumin
Amount is than being 1~3:100.
4. the method as described in claim 1, which is characterized in that in step (1), the temperature of high temperature enzyme deactivation is 90~100 DEG C, when
Between be 10~15min.
5. the method as described in claim 1, which is characterized in that in step (1), the temperature of the primary enzymolysis is 40~60
DEG C, the time is 2~5h.
6. the method as described in claim 1, which is characterized in that in step (1), the ProteAX protease and lactalbumin
Mass ratio be 1~3:100.
7. the method as described in claim 1, which is characterized in that in step (1), the temperature of the secondary enzymolysis is 40~60
DEG C, the time is 2~5h.
8. the method as described in claim 1, which is characterized in that in step (2), the rotating speed of the centrifugation is 6000~8000r/
Min, time are 10~20min.
9. the method as described in claim 1, which is characterized in that in step (2), the interception of the ultrafiltration is 6000~
10000Da, pressure are 0.4~0.6MPa.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111202162A (en) * | 2020-01-20 | 2020-05-29 | 广东蛋能量生物科技有限公司 | Instant egg white peptide sausage and preparation method thereof |
| CN112075529A (en) * | 2020-09-15 | 2020-12-15 | 中国食品发酵工业研究院有限公司 | Method for preparing antioxidant peptide powder by enzymolysis of whey protein powder and application of antioxidant peptide powder in milk tea powder |
| CN112662723A (en) * | 2020-12-30 | 2021-04-16 | 南通励成生物工程有限公司 | Preparation method of debitterized hydrolyzed whey protein rich in small molecular polypeptide |
| CN113229427A (en) * | 2021-03-22 | 2021-08-10 | 武汉名实生物医药科技有限责任公司 | Compound hydrolyzed peptide functional beverage suitable for being drunk by tumor patients and preparation method thereof |
| CN113604528A (en) * | 2021-08-03 | 2021-11-05 | 山东国力生物科技有限公司 | Method for preparing small molecular whey protein active peptide by directional enzyme digestion technology |
| CN113801908A (en) * | 2021-08-23 | 2021-12-17 | 杭州娃哈哈科技有限公司 | Whey protein source functional peptide for relieving physical fatigue and preparation method thereof |
| CN113892655A (en) * | 2021-10-12 | 2022-01-07 | 长春大学 | Method for preparing digestion-free whey protein by taking whey protein powder as raw material |
| CN114540449A (en) * | 2022-03-02 | 2022-05-27 | 江南大学 | High-glutamine content hydrolyzed whey protein powder with improved digestibility and preparation method thereof |
| CN114807282A (en) * | 2022-06-07 | 2022-07-29 | 湖北瑞邦生物科技有限公司 | Rice peptide with low bitter taste and antioxidant and anti-fatigue effects and preparation method thereof |
| CN115530283A (en) * | 2022-09-29 | 2022-12-30 | 黑龙江飞鹤乳业有限公司 | Protein compositions |
| CN116268174A (en) * | 2023-03-10 | 2023-06-23 | 华南理工大学 | Low-bitter casein zymolyte and preparation method and application thereof |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111202162A (en) * | 2020-01-20 | 2020-05-29 | 广东蛋能量生物科技有限公司 | Instant egg white peptide sausage and preparation method thereof |
| CN112075529A (en) * | 2020-09-15 | 2020-12-15 | 中国食品发酵工业研究院有限公司 | Method for preparing antioxidant peptide powder by enzymolysis of whey protein powder and application of antioxidant peptide powder in milk tea powder |
| CN112662723A (en) * | 2020-12-30 | 2021-04-16 | 南通励成生物工程有限公司 | Preparation method of debitterized hydrolyzed whey protein rich in small molecular polypeptide |
| CN113229427A (en) * | 2021-03-22 | 2021-08-10 | 武汉名实生物医药科技有限责任公司 | Compound hydrolyzed peptide functional beverage suitable for being drunk by tumor patients and preparation method thereof |
| CN113604528B (en) * | 2021-08-03 | 2023-11-17 | 山东国力生物科技有限公司 | Method for preparing micromolecular whey protein active peptide by directional enzyme digestion technology |
| CN113604528A (en) * | 2021-08-03 | 2021-11-05 | 山东国力生物科技有限公司 | Method for preparing small molecular whey protein active peptide by directional enzyme digestion technology |
| CN113801908A (en) * | 2021-08-23 | 2021-12-17 | 杭州娃哈哈科技有限公司 | Whey protein source functional peptide for relieving physical fatigue and preparation method thereof |
| CN113801908B (en) * | 2021-08-23 | 2024-04-09 | 杭州娃哈哈科技有限公司 | Whey protein source functional peptide for relieving physical fatigue and preparation method thereof |
| CN113892655A (en) * | 2021-10-12 | 2022-01-07 | 长春大学 | Method for preparing digestion-free whey protein by taking whey protein powder as raw material |
| CN114540449A (en) * | 2022-03-02 | 2022-05-27 | 江南大学 | High-glutamine content hydrolyzed whey protein powder with improved digestibility and preparation method thereof |
| CN114540449B (en) * | 2022-03-02 | 2024-04-02 | 江南大学 | Hydrolyzed whey protein powder with improved digestibility and high glutamine content and preparation thereof |
| CN114807282A (en) * | 2022-06-07 | 2022-07-29 | 湖北瑞邦生物科技有限公司 | Rice peptide with low bitter taste and antioxidant and anti-fatigue effects and preparation method thereof |
| CN115530283B (en) * | 2022-09-29 | 2024-01-26 | 黑龙江飞鹤乳业有限公司 | Protein composition |
| CN115530283A (en) * | 2022-09-29 | 2022-12-30 | 黑龙江飞鹤乳业有限公司 | Protein compositions |
| CN116268174A (en) * | 2023-03-10 | 2023-06-23 | 华南理工大学 | Low-bitter casein zymolyte and preparation method and application thereof |
| CN116268174B (en) * | 2023-03-10 | 2024-04-12 | 华南理工大学 | Low-bitter casein zymolyte and preparation method and application thereof |
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