CN103053788A - Method for preparing enzymatic cross-linked casein by taking yak milk casein as raw material - Google Patents
Method for preparing enzymatic cross-linked casein by taking yak milk casein as raw material Download PDFInfo
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Abstract
The invention relates to a method for preparing enzymatic cross-linked casein by taking yak milk casein as a raw material. The method comprises the following steps of: (1) preparing a casein solution, namely, dissolving the casein in deionized water, performing magnetic stirring till the complete dissolution of the casein, regulating pH value and stabilizing the pH value at 6.5-7.5 to obtain a casein solution; (2) performing enzyme activation: dissolving transglutaminase in the deionized water, and activating in a constant-temperature water bath to obtain an enzyme solution; (3) performing enzymatic cross-linking: adding the enzyme solution into the casein solution, mixing, then performing constant-temperature magnetic stirring reaction to obtain a mixed solution, and performing heat preservation on the mixed solution through a water bath to perform enzyme inactivation to obtain an enzymatic cross-linked casein solution; and (4) obtaining an enzymatic cross-linked casein product: adjusting the pH value of the enzymatic cross-linked casein solution to an isoelectric point, precipitating the casein, centrifuging to remove supernatant fluid to obtain a precipitate, and performing freeze-drying on the precipitate till constant weight to finally obtain the powdered white enzymatic cross-linked casein product. The method disclosed by the invention is easy to control and good in modification effect.
Description
Technical field
The present invention relates to improve protein function character field, relate in particular to a kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material.
Background technology
Yak is Qinghai-Tibet distinctive prepotent sire, approximately 1,500 ten thousand of the existing yaks in the whole world, and China is the main product state of yak, accounts for about 92% of the world total, mainly is distributed in the provinces and regions such as Tibet, Sichuan, Qinghai, Gansu, Xinjiang.Yak lives in the above free of contamination high-cold steppe of height above sea level 3000m for a long time, yak milk is out-and-out natural green food, it also is the topmost living material of Qinghai-xizang Plateau Region herdsman source, except being drunk by the herdsman on a small quantity, all the other major parts are made song and are drawn (Tibetan language: the dried slag of suckling) after degreasing, precipitation, drying.
Yak milk is compared with He Sitan cow's milk, has specificity at aspects such as nutritional labeling, casein structure and compositions.The yak milk dry matter content is 17.86 ~ 18.36%, and is higher by 53% than He Sitan cow's milk (12.00%).The yak milk protein content is 4.62 ~ 5.84%, and casein content is 4.02 %.Yak milk
βThe content very high (greater than 45%) of-casein in total casein,
α s-casein accounts for total caseic 40%,
κ-casein accounts for total caseic 15%, forms with breast milk to approach, and be fit to very much infant's nutritional need, and He Sitan cow's milk is respectively 40%, 50%, 10%.Thereby yak milk is a kind of unique resource that exploitation is worth that has.But the utilization and exploitation of yak milk resource still rests on lower level at present.
Casein (casein, CN) accounts for 80% of Ruzhong total protein.Cow's milk is made Powdered goods and is casein after degreasing, sour precipitation, dehydration, drying, claim again casein.Casein and goods thereof have higher nutritive value and good functional characteristic, are suitable as emulsifying agent, thickener and nutrition fortifier etc., are widely used in food industry, chemical industry and medical industry.Gansu Province is maximum casein production base, the whole nation, and 12 of casein manufacturing enterprises are arranged, and all with yak milk Qu Lawei raw material, produces yak milk casein per year more than 1.35 ten thousand tons, accounts for more than 90% of national casein total output.Yet, because bent preparation of drawing still is in Tibetan area herdsman's extensive style individual workship process segment, cause song to draw the functional characteristics such as physics and chemistry, sense organ and emulsibility, gelling and stability of casein product to be greatly affected.
In order to improve the functional characteristic of yak milk casein, enlarge caseic range of application, adopt the enzyme process cross-linked modification method of appropriateness that its original functional characteristic is improved, be more suitable for the processing of food and chemical industry.Protein under the catalytic action of glutamine transaminage, the γ on the epsilon-amino on the lysine and the glutamic acid-amide groups combination, thus cause protein molecule generation covalent cross-linking, form corresponding polymer.Glutamine transaminage cross-linked proteins reaction condition is gentle, and selectivity is strong, and is cheap, safe, is a kind of common food additive, for food protein is modified first-selected enzyme preparation.The enzymatic modification effect is remarkable, and reaction condition is gentle, and safe, the casein stability after the modification significantly strengthens, and the functional characters such as viscosity, emulsibility, dissolubility, foaminess, film forming are improved.
Summary of the invention
Technical problem to be solved by this invention provide a kind of easy control, modified effect good prepare the crosslinked caseic method of enzyme process take yak milk casein as raw material.
For addressing the above problem, a kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material of the present invention may further comprise the steps:
⑴ prepare casein solution: the feed liquid mass volume ratio of casein by 1:50 ~ 1:150 is dissolved in the deionized water, until casein dissolves fully, and passes through 1.0molL 30 ~ 40 ℃ of magnetic agitation
-1Sodium hydroxide solution regulate the pH value and make it stabilize to 6.5 ~ 7.5, obtain casein solution;
⑵ the activation of enzyme: after being dissolved in the deionized water by the feed liquid mass volume ratio of 1:200 ~ 1.5:200 glutamine transaminage, place 35 ~ 40 ℃ of waters bath with thermostatic control to activate 25 ~ 40min, obtain enzyme solutions;
⑶ enzyme process is crosslinked: be that the ratio of 10 ~ 15U/g adds described enzyme solutions according to glutamine transaminage and caseic proportioning in described casein solution, react 120 ~ 140min at 45 ~ 55 ℃ of temperature constant magnetic stirrings after mixing, obtain mixed liquor, this mixed liquor places 85 ~ 90 ℃ of water-baths to be incubated 5 ~ 10min and makes enzyme deactivation, obtains the crosslinked casein solution of enzyme process;
⑷ the crosslinked Casilan of enzyme process: adopt 1.0 molL
-1Hydrochloric acid solution adjust the crosslinked casein solution pH value of described enzyme process to isoelectric point, make casein precipitation, in the centrifugal 20 ~ 40min of 3000 ~ 4000g centrifugal force, abandoning supernatant, be precipitated thing, this sediment to constant weight, namely gets the crosslinked Casilan of Powdered white enzyme process through freeze drying.
Casein among the described step ⑴ is that yak sweet milk casein, yak milk casein product or yak milk song draw any one in the casein.
The enzyme activity of the glutamine transaminage among the described step ⑵ is 100 U/g.
Cryodesiccated condition refers to that temperature is-40 ~-60 ℃ among the described step ⑷, and pressure is 500 ~ 600Pa, and the time is 20 ~ 30h.
The present invention compared with prior art has the following advantages:
1, the present invention is take yak milk casein as raw material, and glutamine transaminage is crosslinking agent, and reaction condition is gentle, easily control, and modified effect is good, is fit to suitability for industrialized production and popularization, and can be food and chemical industry provides processing stability higher protein batching.
2, to adopt glutamine transaminage be the cross-linking enzyme preparation in the present invention, and this enzyme is a kind of common food additive, and reaction condition is gentle, and selectivity is strong, and is cheap, safe.Protein is under the catalytic action of glutamine transaminage, γ on epsilon-amino on the lysine and the glutamic acid-amide groups combination, thereby cause protein molecule generation covalent cross-linking, form corresponding polymer, increased the active force of stabilize casein micella, casein stability after crosslinked is significantly strengthened, can better satisfy different food products industrial processes demand.
Preparation condition of the present invention, crosslinking degree and crosslinked casein heat endurance are as shown in table 1.Wherein, the stability rate of deposition that is 2% casein solution behind pH=7.0,60 ℃ of heat treatment 30min.
Table 1
As can be seen from Table 1, casein glutamine transaminage cross-linking reaction mild condition, each conditional parameter scope is larger, easily control.Uncrosslinked casein rate of deposition is 5.23, and the more uncrosslinked casein heat endurance of the crosslinked casein of enzyme process significantly improves.
Embodiment 1In, the more uncrosslinked casein of crosslinked casein heat endurance has improved 1.25 times.
Embodiment 2In, the more uncrosslinked casein of crosslinked casein heat endurance has improved 1.33 times.
Embodiment 3In, the more uncrosslinked casein of crosslinked casein heat endurance has improved 1.34 times.Therefore, crosslinked action can significantly improve caseic heat endurance, can better meet food and chemical industry process requirements.
3, the present invention is take yak milk casein goods such as yak milk casein, yak milk Qu La as raw material, the crosslinked casein of preparation glutamine transaminage, the poor technical problems of functional character such as yak milk casein stability have been overcome, improved the added value of yak milk casein, having enlarged it should scope.
The specific embodiment
Embodiment 1A kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material may further comprise the steps:
⑴ prepare casein solution: the feed liquid mass volume ratio (g/mL) of casein by 1:50 is dissolved in the deionized water, until casein dissolves fully, and passes through 1.0molL 30 ℃ of magnetic agitation
-1Sodium hydroxide solution regulate the pH value and make it stabilize to 6.5, obtain casein solution.
⑵ the activation of enzyme: after being dissolved in the deionized water by the feed liquid mass volume ratio (g/mL) of 1:200 glutamine transaminage, place 35 ℃ of waters bath with thermostatic control to activate 25min, obtain enzyme solutions.
⑶ enzyme process is crosslinked: be that the ratio of 10U/g adds enzyme solutions according to glutamine transaminage and caseic proportioning in casein solution, react 120min at 45 ℃ of temperature constant magnetic stirrings after mixing, obtain mixed liquor, this mixed liquor places 85 ℃ of water-bath insulation 10min to make enzyme deactivation, obtains the crosslinked casein solution of enzyme process.
⑷ the crosslinked Casilan of enzyme process: adopt 1.0 molL
-1Hydrochloric acid solution adjust the crosslinked casein solution pH value of enzyme process to isoelectric point, make the casein precipitation, in the centrifugal 40min of 3000g centrifugal force, abandoning supernatant, be precipitated thing, this sediment is that freeze drying 30h namely gets the crosslinked Casilan of Powdered white enzyme process to constant weight under the condition of 500Pa for-40 ℃, pressure in temperature.
Embodiment 2A kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material may further comprise the steps:
⑴ prepare casein solution: the feed liquid mass volume ratio (g/mL) of casein by 1:150 is dissolved in the deionized water, until casein dissolves fully, and passes through 1.0molL 40 ℃ of magnetic agitation
-1Sodium hydroxide solution regulate the pH value and make it stabilize to 7.5, obtain casein solution.
⑵ the activation of enzyme: after being dissolved in the deionized water by the feed liquid mass volume ratio (g/mL) of 1.5:200 glutamine transaminage, place 40 ℃ of waters bath with thermostatic control to activate 40min, obtain enzyme solutions.
⑶ enzyme process is crosslinked: be that the ratio of 15U/g adds enzyme solutions according to glutamine transaminage and caseic proportioning in casein solution, react 140min at 55 ℃ of temperature constant magnetic stirrings after mixing, obtain mixed liquor, this mixed liquor places 90 ℃ of water-bath insulation 5min to make enzyme deactivation, obtains the crosslinked casein solution of enzyme process.
⑷ the crosslinked Casilan of enzyme process: adopt 1.0 molL
-1Hydrochloric acid solution adjust the crosslinked casein solution pH value of enzyme process to isoelectric point, make the casein precipitation, in the centrifugal 20min of 4000g centrifugal force, abandoning supernatant, be precipitated thing, this sediment is that freeze drying 20h namely gets the crosslinked Casilan of Powdered white enzyme process to constant weight under the condition of 600Pa for-60 ℃, pressure in temperature.
Embodiment 3A kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material may further comprise the steps:
⑴ prepare casein solution: the feed liquid mass volume ratio (g/mL) of casein by 1:100 is dissolved in the deionized water, until casein dissolves fully, and passes through 1.0molL 35 ℃ of magnetic agitation
-1Sodium hydroxide solution regulate the pH value and make it stabilize to 7.0, obtain casein solution.
⑵ the activation of enzyme: after being dissolved in the deionized water by the feed liquid mass volume ratio (g/mL) of 1.3:200 glutamine transaminage, place 37 ℃ of waters bath with thermostatic control to activate 30min, obtain enzyme solutions.
⑶ enzyme process is crosslinked: be that the ratio of 13U/g adds enzyme solutions according to glutamine transaminage and caseic proportioning in casein solution, react 130min at 50 ℃ of temperature constant magnetic stirrings after mixing, obtain mixed liquor, this mixed liquor places 88 ℃ of water-bath insulation 7min to make enzyme deactivation, obtains the crosslinked casein solution of enzyme process.
⑷ the crosslinked Casilan of enzyme process: adopt 1.0 molL
-1Hydrochloric acid solution adjust the crosslinked casein solution pH value of enzyme process to isoelectric point, make the casein precipitation, in the centrifugal 30min of 3500g centrifugal force, abandoning supernatant, be precipitated thing, this sediment is that freeze drying 25h namely gets the crosslinked Casilan of Powdered white enzyme process to constant weight under the condition of 550Pa for-50 ℃, pressure in temperature.
Above-mentioned
Embodiment 1 ~ 3In casein be that yak sweet milk casein, yak milk casein product or yak milk song draw any one in the casein.The enzyme activity of glutamine transaminage is 100 U/g.
Should be appreciated that, embodiment discussed here and embodiment can propose various modifications and variations just in order to illustrate to the people who is familiar with this field, and these improvement and variation will be included in the application's Spirit Essence and scope and the appended claim scope.
Claims (4)
1. one kind prepares the crosslinked caseic method of enzyme process take yak milk casein as raw material, may further comprise the steps:
⑴ prepare casein solution: the feed liquid mass volume ratio of casein by 1:50 ~ 1:150 is dissolved in the deionized water, until casein dissolves fully, and passes through 1.0molL 30 ~ 40 ℃ of magnetic agitation
-1Sodium hydroxide solution regulate the pH value and make it stabilize to 6.5 ~ 7.5, obtain casein solution;
⑵ the activation of enzyme: after being dissolved in the deionized water by the feed liquid mass volume ratio of 1:200 ~ 1.5:200 glutamine transaminage, place 35 ~ 40 ℃ of waters bath with thermostatic control to activate 25 ~ 40min, obtain enzyme solutions;
⑶ enzyme process is crosslinked: be that the ratio of 10 ~ 15U/g adds described enzyme solutions according to glutamine transaminage and caseic proportioning in described casein solution, react 120 ~ 140min at 45 ~ 55 ℃ of temperature constant magnetic stirrings after mixing, obtain mixed liquor, this mixed liquor places 85 ~ 90 ℃ of water-baths to be incubated 5 ~ 10min and makes enzyme deactivation, obtains the crosslinked casein solution of enzyme process;
⑷ the crosslinked Casilan of enzyme process: adopt 1.0 molL
-1Hydrochloric acid solution adjust the crosslinked casein solution pH value of described enzyme process to isoelectric point, make casein precipitation, in the centrifugal 20 ~ 40min of 3000 ~ 4000g centrifugal force, abandoning supernatant, be precipitated thing, this sediment to constant weight, namely gets the crosslinked Casilan of Powdered white enzyme process through freeze drying.
2. a kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material as claimed in claim 1, it is characterized in that: the casein among the described step ⑴ is that yak sweet milk casein, yak milk casein product or yak milk song draw any one in the casein.
3. a kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material as claimed in claim 1, it is characterized in that: the enzyme activity of the glutamine transaminage among the described step ⑵ is 100 U/g.
4. a kind of crosslinked caseic method of enzyme process for preparing take yak milk casein as raw material as claimed in claim 1, it is characterized in that: cryodesiccated condition refers to that temperature is-40 ~-60 ℃ among the described step ⑷, and pressure is 500 ~ 600Pa, and the time is 20 ~ 30h.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105506046A (en) * | 2016-01-15 | 2016-04-20 | 中国农业大学 | Yak milk Qula based casein glucose-lowering lipid-lowering peptide as well as preparation method and application thereof |
| CN109295137A (en) * | 2018-10-17 | 2019-02-01 | 四川大学 | A kind of method that the continuous enzymatic microfibrillar collagen of multistep is cross-linking modified |
| CN112753770A (en) * | 2020-12-28 | 2021-05-07 | 西藏农牧学院 | Processing technology for improving stability of yak yoghourt |
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2013
- 2013-01-28 CN CN2013100316495A patent/CN103053788A/en active Pending
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506046A (en) * | 2016-01-15 | 2016-04-20 | 中国农业大学 | Yak milk Qula based casein glucose-lowering lipid-lowering peptide as well as preparation method and application thereof |
| CN105506046B (en) * | 2016-01-15 | 2019-02-26 | 中国农业大学 | Casein hypoglycemic peptide based on yak milk Qula and its preparation method and application |
| CN109295137A (en) * | 2018-10-17 | 2019-02-01 | 四川大学 | A kind of method that the continuous enzymatic microfibrillar collagen of multistep is cross-linking modified |
| CN112753770A (en) * | 2020-12-28 | 2021-05-07 | 西藏农牧学院 | Processing technology for improving stability of yak yoghourt |
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Application publication date: 20130424 |