CN109875038A - A kind of method, anti-oxidant product and application preparing anti-oxidant product using lactalbumin and lactose - Google Patents
A kind of method, anti-oxidant product and application preparing anti-oxidant product using lactalbumin and lactose Download PDFInfo
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Abstract
The present invention provides a kind of method, anti-oxidant product and applications that anti-oxidant product is prepared using lactalbumin and lactose, it is related to anti-oxidant product preparation technical field, the preparation method includes: that lactalbumin and lactose are carried out Maillard reaction, obtains anti-oxidant product.Using method provided by the invention, the DPPH free radical scavenging activity for the anti-oxidant product being prepared compared with the DPPH free radical scavenging activity of lactalbumin, improves 30% or more 76% or more.
Description
Technical field
The present invention relates to anti-oxidant product preparation technical fields, and in particular to a kind of prepared using lactalbumin and lactose is resisted
The method of oxidation product, anti-oxidant product and application.
Background technique
Free radical is that the one kind generated in organism metabolic processes has the molecule of one or several unpaired electronics
Or atom, chemical property is quite active and has the strong oxidizing property of height.The presence of internal excessive free radicals is to cause carefully
The destruction of born of the same parents and institutional framework, cause function damage, cause the major reason of the diseases such as heart disease, cancer and aging.High activity
The intake of antioxidant is a kind of effective ways for removing internal excessive free radicals.Since artificial synthesized antioxidant exists obviously
Toxic side effect and safety issue, be subject to certain restrictions in use, therefore it is active natural how to find high anti-oxidation
Ingredient has become the research hotspot of functional food field.
Summary of the invention
The purpose of the present invention is to provide a kind of methods, anti-oxidant that anti-oxidant product is prepared using lactalbumin and lactose
Lactalbumin and lactose are carried out Maillard reaction, can obtain the active product of high anti-oxidation by product and application, the present invention.
The present invention provides a kind of methods for preparing anti-oxidant product using lactalbumin and lactose, comprising: by whey egg
White and lactose carries out Maillard reaction, obtains anti-oxidant product.
Preferably, the mass ratio of the lactalbumin and lactose is 1:1~5.
Preferably, the temperature of the Maillard reaction is 110~115 DEG C.
Preferably, the temperature of the Maillard reaction is 110.73 DEG C.
Preferably, the time of the Maillard reaction is 1.75~2.2h.
Preferably, the time of the Maillard reaction is 1.93h.
Preferably, the pH value of the Maillard reaction is 11~12.
Preferably, the pH value of the Maillard reaction is 11.51.
The present invention also provides a kind of anti-oxidant product, anti-oxidant product preparation side as described in above-mentioned technical proposal
Method is prepared.
The present invention also provides application of the anti-oxidant product in food described in above-mentioned technical proposal.
The present invention provides a kind of the method for anti-oxidant product, anti-oxidant product are prepared using lactalbumin and lactose and answer
With the preparation method includes: that lactalbumin and lactose are carried out Maillard reaction, obtains anti-oxidant product.In the present invention,
Covalent polymerization reaction occurs for the lactalbumin and lactose, and the antioxidant activity of the anti-oxidant product made greatly improves.
The embodiment of the present invention is as the result is shown: method provided by the invention is used, the anti-oxidant product being prepared
DPPH free radical scavenging activity is 76% or more, compared with the DPPH free radical scavenging activity of lactalbumin, improve 30% with
On.
Detailed description of the invention
Fig. 1 is the anti-oxidant product gel electrophorogram of whey wastewater, wherein 1. protein standard substances；2. whey wastewater；3. useless
Aquatic products；α-La, alpha lactalbumin；β-Lg, beta lactoglobulin；
Fig. 2 is infrared spectrum scanning result.
Specific embodiment
The present invention provides a kind of methods for preparing anti-oxidant product using lactalbumin and lactose, comprising: by whey egg
White and lactose carries out Maillard reaction, obtains anti-oxidant product.
In the present invention, the mass ratio of the lactalbumin and lactose is preferably 1:1~5.In the present invention, the whey
The mass ratio of albumen and lactose is outside above range, with the increase of mass ratio.The anti-oxidant product being prepared it is anti-oxidant
Activity gradually lowers, and the browning degree of anti-oxidant product gradually increases, and is due to increasing the reaction density of lactose, reaction rate
Accelerate, generates more insoluble substances, antioxidant activity is caused to reduce；With the reduction of mass ratio, antioxidant activity is reduced
It is not significant.
In the present invention, the temperature of the Maillard reaction is preferably 110~115 DEG C, and more preferably 110.73 DEG C.At this
In invention, antioxidant activity of the preparation temperature of anti-oxidant product in the range of 110~115 DEG C is most strong, 115 DEG C it
Afterwards, as the temperature increases, the molecular weight of the anti-oxidant product of generation increases, and does not have or have lower antioxidant activity.
In the present invention, the time of the Maillard reaction is preferably 1.75~2.2h, more preferably 1.93h.In this hair
In bright, in the range of 1.75~2.2h, antioxidant activity highest is prepared the preparation time of the anti-oxidant product after 2.2h
The antioxidant activity of obtained anti-oxidant product is not significant.
In the present invention, the pH value of the Maillard reaction is 11~12, preferably 11.51.In the present invention, described anti-
For the preparation pH value of oxidation product at 11~12, the antioxidant activity highest of anti-oxidant product, pH value is after 12, anti-oxidant production
The antioxidant activity of object reduces.
In the present invention, the lactalbumin and lactose are preferably derived from yak whey wastewater, the yak whey wastewater
From Part of Qinghai Plateau pastoral song dairy industry Co., Ltd.In the present invention, a large amount of fat, junket are contained in the yak whey wastewater
The impurity such as albumen and the lactalbumin of denaturation, it is necessary to remove these impurities, lactalbumin in the yak whey wastewater after removal
Content be preferably 5~6.5g/L, more preferably 5.24g/L；The content of lactose is preferably 4~5g/L, more preferably 4.53g/
L。
The present invention also provides a kind of anti-oxidant product, anti-oxidant product preparation side as described in above-mentioned technical proposal
Method is prepared.
The present invention also provides application of the anti-oxidant product in food described in above-mentioned technical proposal.
Combined with specific embodiments below a kind of anti-oxidant product is prepared using lactalbumin and lactose to of the present invention
Method, anti-oxidant product and application be further described in detail, technical solution of the present invention includes but is not limited to following real
Apply example.
Embodiment 1
Yak whey wastewater derives from Part of Qinghai Plateau pastoral song dairy industry Co., Ltd, because in untreated yak whey wastewater
Containing impurity such as a large amount of fat, casein and the lactalbumins of denaturation, these impurity can generate follow-up test and seriously affect,
Therefore whey wastewater must be pre-processed before testing.The present embodiment adjusts yak with 0.5mol/LHCl and 0.5mol/LNaOH
After the pH value of bovine whey waste water is 4.6, at room temperature with 11000rpm high speed centrifugation 10min grease removal removal of impurities, take supernatant spare；
Protein content of whey is 0.600g, lactose content 0.453g in the supernatant of 100mL, adjusts whey in supernatant
The mass ratio of albumen and lactose is 1:1.5, and the pH value that supernatant is arranged is 11.51, and the time of Maillard reaction is 1.93h, instead
Answering temperature is 110.73 DEG C, after reaction by reaction solution 11000rpm high speed centrifugation 15min, measures anti-oxidant production in supernatant
The DPPH clearance rate of object；
The above-mentioned supernatant of 10.0mL is respectively taken, constant volume to 50mL takes constant volume liquid 1.0mL, and sequentially adding mass concentration is
The DPPH ethanol solution of 0.025g/L, shakes up, is protected from light 25min immediately, and absorbance A is surveyed at 517nmX.Similarly,
It gets 1.0mL and determines solution, the DPPH ethanol solution mixing of 4.0mL 0.025g/L is added as control, absorbance is
A0；Make reference, absorbance A with 1.0mL distilled water and 4.0mL dehydrated alcohol mixed solutionx0Beauty is calculated as follows to draw
The antioxidant activity of moral reaction product, i.e. DPPH free radical scavenging activity.
A in formulaXThe absorbance after balance is reacted with DPPH ethanol solution for sample-adding product；
AX0For the sample background absorbance for not adding DPPH ethanol solution；
A0For the absorbance for not being loaded product DPPH ethanol solution.
The calculation shows that, the DPPH free radical scavenging activity of anti-oxidant product is 78.77% in supernatant.
Embodiment 2
The source of yak whey wastewater and processing mode are with embodiment 1, and protein content of whey is in the supernatant of 100mL
0.6g, lactose content 0.453g, adjusting the mass ratio of lactalbumin and lactose in supernatant is 1:1.5, and supernatant is arranged
PH value is 11.5, and the time of Maillard reaction is 1.9h, and reaction temperature is 111 DEG C, after reaction by reaction solution 11000rpm
High speed centrifugation 15min measures the DPPH clearance rate of anti-oxidant product in supernatant, and measuring method is the same as embodiment 1.
The calculation shows that, the DPPH free radical scavenging activity of anti-oxidant product is 76.12% in supernatant.
Comparative example 1
The source of yak whey wastewater and processing mode are with embodiment 1, and protein content of whey is in the supernatant of 100mL
0.6g measures the DPPH clearance rate of lactalbumin in supernatant, and measuring method is the same as embodiment 1.
The calculation shows that, the DPPH free radical scavenging activity of the lactalbumin in supernatant are 44.73%.
Embodiment 3
The source of yak whey wastewater and processing mode are with embodiment 1, and protein content of whey is in the supernatant of 100mL
0.6g, lactose content 0.453g, adjusting the mass ratio of lactalbumin and lactose in supernatant is 1:5, and the pH of supernatant is arranged
Value is 11, and the time of Maillard reaction is 1.75h, and reaction temperature is 110 DEG C, after reaction by reaction solution 11000rpm high speed
It is centrifuged 15min, measures the DPPH clearance rate of anti-oxidant product in supernatant, measuring method is the same as embodiment 1.
The calculation shows that, the DPPH free radical scavenging activity of anti-oxidant product is 76.48% in supernatant.
Embodiment 4
The source of yak whey wastewater and processing mode are with embodiment 1, and protein content of whey is in the supernatant of 100mL
0.6g, lactose content 0.453g, adjusting the mass ratio of lactalbumin and lactose in supernatant is 1:1, and the pH of supernatant is arranged
Value is 12, and the time of Maillard reaction is 2.2h, and reaction temperature is 115 DEG C, after reaction by reaction solution 11000rpm high speed
It is centrifuged 15min, measures the DPPH clearance rate of anti-oxidant product in supernatant, measuring method is the same as embodiment 1.
The calculation shows that, the DPPH free radical scavenging activity of anti-oxidant product is 76.82% in supernatant.
Embodiment 5
Reaction solution after 2 Maillard reaction of the Example high speed centrifugation 7min at 11000rpm, removes insoluble
Maillard reaction product and impurity take 2 times of supernatant dilution spare.Use 15% separation gel, 5% concentration glue, applied sample amount 8
μ L adjusts voltage to 60V, can start electrophoresis.When bromophenol blue indicator enters separation gel forward position, voltage is adjusted to 120V.Work as bromine
When phenol indigo plant indicator Front distance separation gel bottom end about 1.5cm, terminate electrophoresis, immobilized gel 0.5h takes out, and uses Coomassie brilliant blue
Dye 60 DEG C of dyeing 6min, it is clear that destainer is eluted to protein band, takes pictures, exports, in case analysis.Electrophoresis is to use not connect
Continuous buffer system carries out SDS-PAGE electrophoresis.The result is shown in Figure 1.
As can be drawn from Figure 1, band 1 is protein standard substance characteristic bands, and range of molecular weight distributions is 11~180KD；
Band 2 is lactalbumin band in unreacted whey wastewater, is respectively alpha lactalbumin, β-milk-globule egg in the band of 9KD, 14KD
It is white；Band 3 is the anti-oxidant product band of whey wastewater.By electrophorogram in Fig. 1 it is found that reaction after alpha lactalbumin and β-milk-globule
Molecular weight of albumen increases respectively to 10KD and 16KD or so, this phenomenon and Yuan's denier et al. are using WPI and lactose in temperature 70 C, pH
7.0, react 12 under the conditions of albumen and lactose ratio 1:1~result that obtains for 24 hours is consistent；Result of study shows the cream in whey wastewater
Albumin and lactose have it is covalent polymerize, molecular weight all increased, and increase by 2~3KD or so.
Embodiment 6
Reaction solution after 2 Maillard reaction of Example obtains supernatant in 11000rpm high speed centrifugation 7min.First use
The bag filter of 10KD, which is dialysed to supernatant fluid color, to be basically unchanged, and lactose and impurity that bag filter middle-molecular-weihydroxyethyl is less than 10KD are removed,
Dialyzate is discarded, trapped fluid is retained, replaces dialyzate every 4h.Then, it is similarly dialysed again with the bag filter of 20KD to trapped fluid
Dialyzate color is basically unchanged, and discards trapped fluid, retains dialyzate, and 45~50 DEG C of rotary evaporations of dialyzate of 10~20KD are dense
Contracting, freeze-drying, to obtain the anti-oxidant product albumen powder that molecular weight is 10~20KD.It is 10~20KD antioxygen by molecular weight
Change PURE WHEY KBr tabletting obtained by product albumen powder and foamet, carry out infrared spectrum scanning, exports data, analysis
As a result map is shown in Fig. 2.
As can be drawn from Figure 2, the infared spectrum of comparison maillard reaction product and foam fraction factor lactalbumin is it is found that beauty
Maillard reaction product is in 3750~3000cm-1Absorption peak broaden by force, 1720cm-1C=O (carbonyl) frequency at place is reduced to
1650cm-1, this illustrates that graft reaction has occurred in lactalbumin in whey wastewater and lactose, this and the anti-oxidant product of whey wastewater
Gel electrophoresis analysis conclusion is consistent.In 3750~3000cm-1Absorption peak broaden by force be because are as follows: lactose has multiple-OH (hydroxyls
Base), the access of lactose increases the-OH content of lactalbumin；Originally there are the presence of amido bond, 1720cm in lactalbumin-1Place
The absorption peak for being absorbed as amido bond in lactalbumin, amido bond is more, and absorption frequency is lower, and the frequency of C=O has been here
Reduction is because of lactalbumin-NH2After lone pair electrons, with lactose C=O graft reaction, there are more amido bonds to generate,
It can cause more albumen-NH2With the conjugation of lactose C=O, shift C=O double bond electronics to C-N singly-bound, C=O
Double bond cloud density reduces, and force constant reduces.
By above embodiments it can be concluded that, using method provided by the invention, the anti-oxidant product that is prepared
DPPH free radical scavenging activity is 76% or more, compared with the DPPH free radical scavenging activity of lactalbumin, improve 30% with
On.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method for preparing anti-oxidant product using lactalbumin and lactose, comprising: lactalbumin and lactose are carried out beauty
Maillard reaction obtains anti-oxidant product.
2. the method according to claim 1, wherein the mass ratio of the lactalbumin and lactose is 1:1~5.
3. the method according to claim 1, wherein the temperature of the Maillard reaction is 110~115 DEG C.
4. according to the method described in claim 3, it is characterized in that, the temperature of the Maillard reaction is 110.73 DEG C.
5. method according to claim 1 or 3, which is characterized in that the time of the Maillard reaction is 1.75~2.2h.
6. according to the method described in claim 5, it is characterized in that, the time of the Maillard reaction is 1.93h.
7. the method according to claim 1, wherein the pH value of the Maillard reaction is 11~12.
8. the method according to the description of claim 7 is characterized in that the pH value of the Maillard reaction is 11.51.
9. a kind of anti-oxidant product, which is characterized in that the anti-oxidant product is by preparation according to any one of claims 1 to 8
Method is prepared.
10. the anti-oxidant product or according to any one of claims 8 anti-that any one of claim 1~8 preparation method is prepared
Application of the oxidation product in food.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102628073A (en) * | 2012-03-28 | 2012-08-08 | 东北农业大学 | Glycosylated lactalbumin polypeptide with antioxidant activity as well as preparation method and application of glycosylated lactalbumin polypeptide |
| CN103304657A (en) * | 2012-03-16 | 2013-09-18 | 东北林业大学 | Glycosylated modifying method for improving antioxidant activity of whey protein |
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