CN102863526A - Separation method of yak milk milk fat globule membrane protein - Google Patents
Separation method of yak milk milk fat globule membrane protein Download PDFInfo
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Abstract
The invention discloses a separation method of yak milk milk fat globule membrane protein, relates to a separation method of protein and solves problems of low purity and unstable properties in terms of cow's milk milk fat globule membrane protein prepared by existing methods. The separation method includes that cane sugar is added in yak milk, a milk fat initial extract is obtained through centrifugation, then centrifugal washing is performed with a sodion phosphate buffer solution and ultrapure water, and finally the centrifugation is performed with a polyethylene glycol octyl phenyl ether solution to obtain the yak milk milk fat globule membrane protein. The prepared yak milk milk fat globule membrane protein has the advantages that the properties are stable, the purity can reach to 93.5%, and problems of low purity and unstable properties in terms of the cow's milk milk fat globule membrane protein prepared by existing separation methods are solved. The separation method is applicable to mass production of the yak milk milk fat globule membrane protein.
Description
Technical field
The present invention relates to a kind of separation method of albumen.
Background technology
Dairy fats is one of main component of cow's milk, dairy fats is scattered in the cow's milk with the form of Oil globule, and its surface is covered by the film of a layer thickness by 5 ~ 10nm, accounts for 2%～6% of Oil globule quality, be called milk fat ball film (Milk Fat Globule Menbrane, MFGM).Milk fat ball film is comprised of protein, phosphatide, glycoprotein, triacylglycerol, cholesterol, enzyme and other trace ingredients; wherein; take glycoprotein as main, milk fat ball film albumen accounts for the 25%-60% of milk fat ball film total mass in the milk fat ball film albumen, accounts for the 1%-2% of total milk protein total mass.Studies show that milk fat ball film albumen has good antitumour activity, the breast cancer susceptibility gene that extracts in the milk fat ball film egg has the effect that suppresses mammary cancer.The separation method of existing milk fat ball film albumen mainly contains acid deposition method and membrane filter method.The acid deposition method is by using salt acid deposition milk fat ball film albumen, the advantage of the method be simple to operate, cost is low, but hydrochloric acid can affect the milk fat ball film protein-active to a certain extent, and it is on the low side and character is unstable to cause separating the purity of the milk fat ball film albumen that obtains; The milk fat ball film property of protein that membrane filter method obtains is better, but the cost of the method is high, and complicated operation is not suitable for a large amount of productions of milk fat ball film albumen.
Summary of the invention
The present invention is the low and unsettled problem of character of purity that will solve the Cow Milk fat globule membrane albumen of existing method preparation, and a kind of separation method of yak milk milk fat ball film albumen is provided.
The separation method of a kind of yak milk milk fat ball film albumen of the present invention carries out according to the following steps:
One, takes by weighing 100 parts yak milk and 3 ~ 8 parts sucrose by mass fraction;
Two, after the yak milk that step 1 is taken by weighing filters with 3 ~ 5 layers of gauze, add the sucrose that step 1 takes by weighing, mix, obtain cow's milk sucrose mixed solution;
Three, the cow's milk sucrose mixed solution that step 2 is obtained carries out centrifugal at ambient temperature, obtains being in the dairy fats primary extract on centrifugal product upper strata; Wherein, centrifugal rotational speed is 3000 ~ 4000r/min, and centrifugation time is 20 ~ 35min;
Four, in the dairy fats primary extract that step 3 obtains, add 1 ~ 3 times to the sodium ion phosphate buffer soln of this dairy fats primary extract volume, after mixing, carry out at ambient temperature centrifuge washing; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min; The compound method of described sodium ion phosphate buffer soln is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.2 ~ 4.4mmol, KCl:2.5 ~ 2.8mmol, KH
2PO
4: the ratio of 1.6 ~ 1.8mmol and NaCl:135～140mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.2 ~ 7.5, obtain the sodium ion phosphate buffer soln;
Five, the dairy fats primary extract that will process through step 4 by the method repetitive operation of step 4 once, obtains being in the dairy fats on centrifugal product upper strata;
Six, in the dairy fats that step 5 obtains, add the ultrapure water identical with the dairy fats volume, after mixing, carry out at ambient temperature centrifuge washing, obtain being in the cream on centrifugal product upper strata; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min;
Seven, the cream that step 6 is obtained places under 2 ~ 6 ℃ of conditions, and leaves standstill 12 ~ 16h under this condition, obtains the cream solid;
Eight, the cream solid that step 7 is obtained places under 40 ~ 45 ℃ the water bath condition and melts, and obtains liquid cream;
Nine, in the liquid cream that step 8 obtains, add volume for accounting for the Triton X-100 solution of liquid state milk oil volume 0.15 ~ 0.3%, carry out at ambient temperature centrifugally, collecting precipitation obtains yak milk milk fat ball film albumen; Wherein, centrifugal rotational speed is 40000 ~ 50000r/min, and centrifugation time is 40 ~ 70min; The compound method of described Triton X-100 solution is: the volume ratio by Triton X-100 and phosphate buffered saline buffer is that 1:2.5 ~ 2.7 take by weighing Triton X-100 and phosphate buffered saline buffer, after the two is mixed, place 37 ~ 40 ℃ water-bath, kept 2 ~ 3 hours; The concentration of phosphate buffered saline buffer is that 0.07 ~ 0.1mol/L, pH are 7.2 ~ 7.4.
The present invention at first adds sucrose in yak cow's milk, can effectively reduce the loss of milk fat ball film albumen in subsequent separation process; Utilize the sodium ion phosphate buffer soln that yak milk dairy fats primary extract is carried out wash-out, sodium ion phosphate buffer soln used in the present invention can wash away casein and the whey-protein that is attached on the milk fat ball film, has reduced the content of impurity albumen in the milk fat ball film albumen that extracts; Utilize nonionic detergent Triton X-100 solution that yak milk cream is carried out wash-out, employed nonionic detergent Triton X-100 solution can lipin dissolving and the character of albumen itself is not impacted, and makes the stable in properties of albumen.The purity to 93 of the yak Cow Milk fat globule membrane albumen by the inventive method preparation ~ 95%, and the purity of the yak Cow Milk fat globule membrane albumen of existing method preparation only is about 80%, effectively raises the purity of yak Cow Milk fat globule membrane albumen.
The present invention is applicable to a large amount of productions of yak milk milk fat ball film albumen.
Embodiment
Technical scheme of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: the separation method of the yak milk milk fat ball film albumen of present embodiment carries out according to the following steps:
One, takes by weighing 100 parts yak milk and 3 ~ 8 parts sucrose by mass fraction;
Two, after the yak milk that step 1 is taken by weighing filters with 3 ~ 5 layers of gauze, add the sucrose that step 1 takes by weighing, mix, obtain cow's milk sucrose mixed solution;
Three, the cow's milk sucrose mixed solution that step 2 is obtained carries out centrifugal at ambient temperature, obtains being in the dairy fats primary extract on centrifugal product upper strata; Wherein, centrifugal rotational speed is 3000 ~ 4000r/min, and centrifugation time is 20 ~ 35min;
Four, in the dairy fats primary extract that step 3 obtains, add 1 ~ 3 times to the sodium ion phosphate buffer soln of this dairy fats primary extract volume, after mixing, carry out at ambient temperature centrifuge washing; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min; The compound method of described sodium ion phosphate buffer soln is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.2 ~ 4.4mmol, KCl:2.5 ~ 2.8mmol, KH
2PO
4: the ratio of 1.6 ~ 1.8mmol and NaCl:135～140mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.2 ~ 7.5, obtain the sodium ion phosphate buffer soln;
Five, the dairy fats primary extract that will process through step 4 by the method repetitive operation of step 4 once, obtains being in the dairy fats on centrifugal product upper strata;
Six, in the dairy fats that step 5 obtains, add the ultrapure water identical with the dairy fats volume, after mixing, carry out at ambient temperature centrifuge washing, obtain being in the cream on centrifugal product upper strata; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min;
Seven, the cream that step 6 is obtained places under 2 ~ 6 ℃ of conditions, and leaves standstill 12 ~ 16h under this condition, obtains the cream solid;
Eight, the cream solid that step 7 is obtained places under 40 ~ 45 ℃ the water bath condition and melts, and obtains liquid cream;
Nine, in the liquid cream that step 8 obtains, add volume for accounting for the Triton X-100 solution of liquid state milk oil volume 0.15 ~ 0.3%, carry out at ambient temperature centrifugally, collecting precipitation obtains yak milk milk fat ball film albumen; Wherein, centrifugal rotational speed is 40000 ~ 50000r/min, and centrifugation time is 40 ~ 70min; The compound method of described Triton X-100 solution is: the volume ratio by Triton X-100 and phosphate buffered saline buffer is that 1:2.5 ~ 2.7 take by weighing Triton X-100 and phosphate buffered saline buffer, after the two is mixed, place 37 ~ 40 ℃ water-bath, kept 2 ~ 3 hours; The concentration of phosphate buffered saline buffer is that 0.07 ~ 0.1mol/L, pH are 7.2 ~ 7.4.
Embodiment two: what present embodiment and embodiment one were different is: take by weighing 20 parts yak milk and 1 part sucrose by mass fraction in the step 1.Other step and parameter are identical with embodiment one.
Embodiment three: what present embodiment was different from embodiment one or two is: centrifugal rotational speed is 3800r/min in the step 3, and centrifugation time is 30min.Other step and parameter are identical with embodiment one or two.
Embodiment four: what present embodiment was different from one of embodiment one to three is: after step 3 and step 4 are centrifugal, centrifugal product is placed under 4 ℃ of conditions, and leave standstill 10min under this condition.Other step and parameter are identical with embodiment one to three.
Embodiment five: what present embodiment was different from one of embodiment one to four is: the compound method of the sodium ion phosphate buffer soln described in the step 4 is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.3mmol, KCl:2.7mmol, KH
2PO
4: the ratio of 1.8mmol and NaCl:137mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.4, obtain the sodium ion phosphate buffer soln.Other step and parameter are identical with embodiment one to four.
Embodiment six: what present embodiment was different from one of embodiment one to five is: centrifugal rotational speed is 4800r/min in step 4 and the step 6, and centrifugation time is 20min.Other step and parameter are identical with embodiment one to five.
Embodiment seven: what present embodiment was different from one of embodiment one to six is: adding volume in the step 9 is the Triton X-100 solution that accounts for liquid state milk oil volume 0.2%.Other step and parameter are identical with embodiment one to six.
Embodiment eight: what present embodiment was different from one of embodiment one to seven is: centrifugal rotational speed is 44910r/min in the step 9, and centrifugation time is 60min.Other step and parameter are identical with embodiment one to seven.
By following verification experimental verification beneficial effect of the present invention:
Test one: the separation method of a kind of yak milk milk fat ball film albumen of this test carries out according to the following steps:
One, selects the fresh yak breast of healthy yak, take by weighing 20 parts yak milk and 1 part sucrose by mass fraction;
Two, after the yak milk that step 1 is taken by weighing filters with 4 layers of gauze, add the sucrose that step 1 takes by weighing, mix with magnetic stirring apparatus, obtain cow's milk sucrose mixed solution;
Three, the cow's milk sucrose mixed solution that step 2 is obtained carries out centrifugal at ambient temperature, centrifugal product is placed under 4 ℃ of conditions, and leave standstill 10min under this condition, obtains being in the dairy fats primary extract on centrifugal product upper strata; Wherein, centrifugal rotational speed is 3800r/min, and centrifugation time is 30min;
Four, in the dairy fats primary extract that step 3 obtains, add 2 times to the sodium ion phosphate buffer soln of this dairy fats primary extract volume, after mixing with magnetic stirring apparatus, carry out at ambient temperature centrifuge washing, centrifugal product is placed under 4 ℃ of conditions, and under this condition, leave standstill 10min; Wherein, centrifugal rotational speed is 4800r/min, and centrifugation time is 20min; The compound method of described sodium ion phosphate buffer soln is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.3mmol, KCl:2.7mmol, KH
2PO
4: the ratio of 1.8mmol and NaCl:137mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.4, obtain the sodium ion phosphate buffer soln;
Five, the dairy fats primary extract that will process through step 4 by the method repetitive operation of step 4 once, obtains being in the dairy fats on centrifugal product upper strata;
Six, in the dairy fats that step 5 obtains, add the ultrapure water identical with the dairy fats volume, after mixing with magnetic stirring apparatus, carry out at ambient temperature centrifuge washing, obtain being in the cream on centrifugal product upper strata; Wherein, centrifugal rotational speed is 4800r/min, and centrifugation time is 20min;
Seven, the cream that step 6 is obtained places under 4 ℃ of conditions, and leaves standstill 15h under this condition, obtains the cream solid;
Eight, the cream solid that step 7 is obtained places under 40 ~ 45 ℃ the water bath condition and melts, and obtains liquid cream;
Nine, in the liquid cream that step 8 obtains, adding volume is the Triton X-100 solution that accounts for liquid state milk oil volume 0.2%, carries out at ambient temperature centrifugally, and collecting precipitation obtains yak milk milk fat ball film albumen; Wherein, centrifugal rotational speed is 44910r/min, and centrifugation time is 60min; The compound method of described Triton X-100 solution is: the volume ratio by Triton X-100 and phosphate buffered saline buffer is that 1:2.6 takes by weighing Triton X-100 and phosphate buffered saline buffer, after the two is mixed, place 37 ℃ water-bath, kept 2 hours; The concentration of phosphate buffered saline buffer is that 0.1mol/L, pH are 7.3.
The controlled trial of test one:
One, selects the fresh yak breast of healthy yak, take by weighing 20 parts yak milk and 1 part sucrose by mass fraction;
Two, after the yak milk that step 1 is taken by weighing filters with 4 layers of gauze, add the sucrose that step 1 takes by weighing, mix with magnetic stirring apparatus, obtain cow's milk sucrose mixed solution;
Three, the cow's milk sucrose mixed solution that step 2 is obtained carries out centrifugal at ambient temperature, centrifugal product is placed under 4 ℃ of conditions, and leave standstill 10min under this condition, obtains being in the dairy fats primary extract on centrifugal product upper strata; Wherein, centrifugal rotational speed is 3800r/min, and centrifugation time is 30min;
Four, in the dairy fats primary extract that step 3 obtains, add 2 times to the sodium ion phosphate buffer soln of this dairy fats primary extract volume, after mixing with magnetic stirring apparatus, carry out at ambient temperature centrifuge washing, centrifugal product is placed under 4 ℃ of conditions, and under this condition, leave standstill 10min; Wherein, centrifugal rotational speed is 4800r/min, and centrifugation time is 20min; The compound method of described sodium ion phosphate buffer soln is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.3mmol, KCl:2.7mmol, KH
2PO
4: the ratio of 1.8mmol and NaCl:137mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.4, obtain the sodium ion phosphate buffer soln;
Five, the dairy fats primary extract that will process through step 4 by the method repetitive operation of step 4 once, obtains being in the dairy fats on centrifugal product upper strata;
Six, in the dairy fats that step 5 obtains, add the ultrapure water identical with the dairy fats volume, after mixing with magnetic stirring apparatus, carry out at ambient temperature centrifuge washing, obtain being in the cream on centrifugal product upper strata; Wherein, centrifugal rotational speed is 4800r/min, and centrifugation time is 20min;
Seven, the cream that step 6 is obtained places under 4 ℃ of conditions, and leaves standstill 15h under this condition, obtains the cream solid;
Eight, the cream solid that step 7 is obtained places under 40 ~ 45 ℃ the water bath condition and melts, and obtains liquid cream;
Nine, the liquid cream that step 8 is obtained stirs 10min with agitator, adds the deionized water identical with the liquid state milk oil volume again, separates obtaining butter and buttermilk;
Ten, the butter that step 9 is obtained is heating and melting in 60 ℃ water-bath, then adds the deionized water identical with the butter volume, is centrifugal 20min under the condition of 4800r/min at rotating speed, discards the upper strata butter, obtains subnatant;
11, the subnatant that step 10 is obtained mixes with the buttermilk that step 10 obtains, be that the HCL of 0.01mol/L regulates its pH to 4.8 with concentration, after leaving standstill 30min under the room temperature condition, be centrifugal 15min under the condition of 12000r/min at rotating speed, collecting precipitation thing and suspended substance namely obtain yak milk milk fat ball film albumen.
By high performance liquid chromatography (HPLC) the yak milk milk fat ball film albumen of testing a yak milk milk fat ball film albumen that obtains and controlled trial and obtaining is analyzed.In the yak milk milk fat ball film albumen that the peak area of impurity peaks only obtains for controlled trial in the test one yak milk milk fat ball film albumen that obtains 60% of the impurity peaks peak area, and assorted peak α
S2The low levels Caseinum componemt of-CN, κ-CN disappears substantially, and yak milk milk fat ball film property of protein is stable.By calculate the test the one yak milk milk fat ball film albumen that obtains purity be 93.5%, and the purity of the yak milk milk fat ball film albumen that controlled trial obtains only is 82.7%, illustrates that the purity of the test one yak milk milk fat ball film albumen that obtains is significantly improved.
Claims (8)
1. the separation method of a yak milk milk fat ball film albumen is characterized in that the separation method of yak milk milk fat ball film albumen carries out according to the following steps:
One, takes by weighing 100 parts yak milk and 3 ~ 8 parts sucrose by mass fraction;
Two, after the yak milk that step 1 is taken by weighing filters with 3 ~ 5 layers of gauze, add the sucrose that step 1 takes by weighing, mix, obtain cow's milk sucrose mixed solution;
Three, the cow's milk sucrose mixed solution that step 2 is obtained carries out centrifugal at ambient temperature, obtains being in the dairy fats primary extract on centrifugal product upper strata; Wherein, centrifugal rotational speed is 3000 ~ 4000r/min, and centrifugation time is 20 ~ 35min;
Four, in the dairy fats primary extract that step 3 obtains, add 1 ~ 3 times to the sodium ion phosphate buffer soln of this dairy fats primary extract volume, after mixing, carry out at ambient temperature centrifuge washing; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min; The compound method of described sodium ion phosphate buffer soln is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.2 ~ 4.4mmol, KCl:2.5 ~ 2.8mmol, KH
2PO
4: the ratio of 1.6 ~ 1.8mmol and NaCl:135 ~ 140mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.2 ~ 7.5, obtain the sodium ion phosphate buffer soln;
Five, the dairy fats primary extract that will process through step 4 by the method repetitive operation of step 4 once, obtains being in the dairy fats on centrifugal product upper strata;
Six, in the dairy fats that step 5 obtains, add the ultrapure water identical with the dairy fats volume, after mixing, carry out at ambient temperature centrifuge washing, obtain being in the cream on centrifugal product upper strata; Wherein, centrifugal rotational speed is 4200 ~ 5200r/min, and centrifugation time is 15 ~ 25min;
Seven, the cream that step 6 is obtained places under 2 ~ 6 ℃ of conditions, and leaves standstill 12 ~ 16h under this condition, obtains the cream solid;
Eight, the cream solid that step 7 is obtained places under 40 ~ 45 ℃ the water bath condition and melts, and obtains liquid cream;
Nine, in the liquid cream that step 8 obtains, add volume for accounting for the Triton X-100 solution of liquid state milk oil volume 0.15 ~ 0.3%, carry out at ambient temperature centrifugally, collecting precipitation obtains yak milk milk fat ball film albumen; Wherein, centrifugal rotational speed is 40000 ~ 50000r/min, and centrifugation time is 40 ~ 70min; The compound method of described Triton X-100 solution is: the volume ratio by Triton X-100 and phosphate buffered saline buffer is that 1:2.5 ~ 2.7 take by weighing Triton X-100 and phosphate buffered saline buffer, after the two is mixed, place 37 ~ 40 ℃ water-bath, kept 2 ~ 3 hours; The concentration of phosphate buffered saline buffer is that 0.07 ~ 0.1mol/L, pH are 7.2 ~ 7.4.
2. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that taking by weighing 20 parts yak milk and 1 part sucrose by mass fraction in the step 1.
3. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that centrifugal rotational speed is 3800r/min in the step 3, and centrifugation time is 30min.
4. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1, it is characterized in that step 3 and step 4 are centrifugal after, centrifugal product is placed under 4 ℃ of conditions, and under this condition, leaves standstill 10min.
5. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that the compound method of the sodium ion phosphate buffer soln described in the step 4 is: by containing Na in every 1L sodium ion phosphate buffer soln
2HPO
4: 4.3mmol, KCl:2.7mmol, KH
2PO
4: the ratio of 1.8mmol and NaCl:137mmol, with Na
2HPO
4, KCl, KH
2PO
4Join in the deionized water with NaCl, and regulate its pH to 7.4, obtain the sodium ion phosphate buffer soln.
6. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that centrifugal rotational speed is 4800r/min in step 4 and the step 6, and centrifugation time is 20min.
7. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that adding volume in the step 9 is the Triton X-100 solution that accounts for liquid state milk oil volume 0.2%.
8. the separation method of a kind of yak milk milk fat ball film albumen according to claim 1 is characterized in that centrifugal rotational speed is 44910r/min in the step 9, and centrifugation time is 60min.
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| CN105455109A (en) * | 2015-12-18 | 2016-04-06 | 光明乳业股份有限公司 | Fat globule membrane powder, preparation method and application thereof |
| CN108299552A (en) * | 2018-02-27 | 2018-07-20 | 吉林大学 | The extracting method of goat dairy milk fat globule membrane protein for proteomics research |
| CN111227039A (en) * | 2018-11-29 | 2020-06-05 | 内蒙古伊利实业集团股份有限公司 | Extraction method of fat globule membrane, yoghourt rich in fat globule membrane and preparation method |
| CN112753770A (en) * | 2020-12-28 | 2021-05-07 | 西藏农牧学院 | Processing technology for improving stability of yak yoghourt |
| CN113892649A (en) * | 2020-06-18 | 2022-01-07 | 湖南农业大学 | Probiotic microcapsule and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105455109A (en) * | 2015-12-18 | 2016-04-06 | 光明乳业股份有限公司 | Fat globule membrane powder, preparation method and application thereof |
| CN108299552A (en) * | 2018-02-27 | 2018-07-20 | 吉林大学 | The extracting method of goat dairy milk fat globule membrane protein for proteomics research |
| CN111227039A (en) * | 2018-11-29 | 2020-06-05 | 内蒙古伊利实业集团股份有限公司 | Extraction method of fat globule membrane, yoghourt rich in fat globule membrane and preparation method |
| CN113892649A (en) * | 2020-06-18 | 2022-01-07 | 湖南农业大学 | Probiotic microcapsule and preparation method thereof |
| CN112753770A (en) * | 2020-12-28 | 2021-05-07 | 西藏农牧学院 | Processing technology for improving stability of yak yoghourt |
| CN112753770B (en) * | 2020-12-28 | 2023-06-16 | 西藏农牧学院 | Processing technology for improving stability of yak yoghourt |
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| CN102863526B (en) | 2014-11-19 |
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