CN1136834C - Method for preparing microencapsulated yeast multienzyme cluster - Google Patents
Method for preparing microencapsulated yeast multienzyme cluster Download PDFInfo
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- CN1136834C CN1136834C CNB01127736XA CN01127736A CN1136834C CN 1136834 C CN1136834 C CN 1136834C CN B01127736X A CNB01127736X A CN B01127736XA CN 01127736 A CN01127736 A CN 01127736A CN 1136834 C CN1136834 C CN 1136834C
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- yeast
- multienzyme
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Abstract
The present invention provides a preparation method for yeast multienzyme complexes and microencapsulated biological products (microencapsulated yeast multienzyme complexes) thereof applicable to the formulas of skin care products, which comprises: fermentation, enzyme production induction, mild extraction, concentration and desiccation, embedding with liposomes and microencapsulation. The method has a reasonable design, the capability of producing highly stable products, the applicability to industrial production and the capability of meeting the need of the market.
Description
The present invention relates to a kind of preparation method of enzyme preparation of micro encapsulation.
At present domestic in skin care item, medicine employed superoxide dismutase SOD be generally the aqueous solution that plant extract or animal blood extract, its homology is poor, the half-life short, enzyme is lived low; Even Liposomal formulation also exists embedding rate low, instability is unfavorable for depositing, shortcoming such as compatibility difference in skin care item prescriptions.
The purpose of this invention is to provide a kind of yeast multienzyme cluster and the biological product of microencapsulation---preparation method of the yeast multienzyme cluster of microencapsulation thereof that is applicable to the skin care item prescription, this biological product mainly are yeast SOD enzymes, peroxidase, bioactive peptide, aminoacid, β-(1,3)-glucosan, oligosaccharide, vitamin, the complex that mineral etc. are formed, form with the liposome of microencapsulation is encapsulated in yeast multienzyme cluster in liposome and the microcapsule bilayer, these goods are as defying age, the agent of antioxidative whitening skin and preserving moisture has good formula compatibility performance and stability.
Technological process of the present invention is as follows:
Fermentation-→ induce the product enzyme-→ gentle the extraction-→ concentrate drying-→ liposome embedded-→ microencapsulation.
Concrete processing step comprises:
(1) preparation yeast multienzyme cluster:
(1.1) strain fermentation:
Strain: adopt bakery yeast [Saccharomyces cerevisiae];
Seed and fermentation medium: molasses fermented culture fluid, high temperature sterilize;
Sweat: by three grades of seed fermentations and be extended to fermentation tank;
Fermentation condition: 25～35 ℃ of temperature, ventilation 0.6: 1～1: 1,20～300 rev/mins of mixing speeds, tank pressure 0.01～0.08MPa, pH keeps 4.0～7.0, ferments to add the hydrogen peroxide of fermentation liquid total amount (weight) 0.001%～0.01% after 4～8 hours, continues fermentation 4～18 hours;
(1.2) after the fermentation ends, centrifugal collection thalline gets yeast paste;
(1.3) yeast paste of gained is handled with isopropyl alcohol, yeast paste: isopropyl alcohol=1: 0.1～20 (weight ratio), filter, get the yeast slag, the phosphate buffer that adds pH5.5～9.0 again, the yeast slag: buffer=1: 0.1～10 (weight ratio) adds lysozyme room temperature simultaneously and handles the centrifuging and taking supernatant;
(1.4) the supernatant cryogenic vacuum with gained concentrates, and promptly gets yeast multienzyme cluster through lyophilization again;
(2) liposome of preparation yeast multienzyme cluster:
(2.1) saturated soybean lecithin, cholesterol, double hexadecyl acid ester are dissolved in ether and make the film material;
(2.2) in order with the mannitol micropowder, the EDTA disodium salt joins in the film material, and decompression stirs;
(2.3) add the yeast multienzyme cluster micropowder, stir;
(2.4) rotary evaporation is removed ether under 20～35 ℃ of temperature, obtains having the liposome powder body of mobile yeast multienzyme cluster;
Each material mixture ratio (percentage by weight) of above-mentioned preparation liposome is: yeast multienzyme cluster 10-80%, saturated soybean lecithin 1-10%, cholesterol 1-10%, EDTA disodium 0.1-0.5%, mannitol 1-5%, double hexadecyl acid ester 0.1-2%;
(3) microencapsulation:
(3.1) with the high molecular polymer be the wall material, the wall material added in the ethanol that mix homogeneously is a wall-forming material film forming solution;
(3.2) with the liposome powder suspension of capsule heartwood material-yeast multienzyme cluster on fluid bed, the control temperature is at 40～50 ℃;
(3.3) with centrifugal spray wall material film forming solution is sprayed in the fluid bed, make dry materials and film forming;
(3.4) sieve classification is crossed in cooling, promptly gets the product of different size.
Live for further improving enzyme, in fermentation technology process of the present invention, but fermentation liquid reuse treatment with uv radiation after adding the hydrogen peroxide processing, promptly in the step in described technological process (1.2), after fermentation ends, again with the fermentation liquid ultraviolet radiation, its dosage is generally 1 * 10
3～1 * 10
6Erg/milliliter, centrifugal again collection thalline after shining with ultraviolet, yeast paste.
Below be relevant optimisation technique feature of the present invention or parameter:
1, in the step in described technological process (1.3), handling yeast paste with isopropyl alcohol, is in yeast paste: the ratio of isopropyl alcohol=1: 1～10 (weight ratio) adds isopropyl alcohol in yeast paste.
2, in the step in described technological process (1.3), the time that yeast paste is handled with isopropyl alcohol is 30 minutes to 120 minutes.
3, in the step in described technological process (1.3) in the yeast slag: the ratio of buffer=1: 1～5 (weight ratio) adds the phosphate buffer of pH7.5～8.5.
4, the amount adding lysozyme room temperature of pressing reaction-ure mixture 0.1～0.01% (percentage by weight) in the step in described technological process (1.3) is handled.
5, in the step in described technological process (1.3), the time of handling the yeast slag with said phosphate buffer and lysozyme is 10 to 20 hours.
6, in the described encapsulation process step, described high molecular polymer wall material is one or more the compositions in ethyl cellulose, PEG-6000, the methylsiloxane monomer.
The present invention has following technical characterstic and effect:
1. the art of this patent is to utilize modern biotechnology fermenting and producing yeast, induce the product enzyme, the multienzyme complex system that is produced is by superoxide dismutase SOD enzyme, peroxidase, bioactive peptide, aminoacid, β-(1,3)-the synergism factors such as glucosan, oligosaccharide, vitamin, mineral, cofactors form, and have very strong radioprotective, antioxidation, the anti-ageing performance of waiting for a long time;
2. handle by gentle, can farthest keep its biological activity;
3. this compound system after the phospholipid embedding, can penetrate cell better, enter cell after, can promote the absorption of cell metabolism, activating cell, promotion oxygen and nutrient;
4. yeast multienzyme cluster can keep its biological activity better after microcapsule embedded, and the yeast multienzyme cluster of microencapsulation not only has good stability, and has slow-release function;
5. the art of this patent technological design is reasonable, is fit to suitability for industrialized production; Properties of product are stable, meeting the market requirement.
Embodiment one
(1) preparation of yeast multienzyme cluster
(1.1) bakery yeast Saccharomyces cerevisiae is activated to three grades of kinds, inserts fermentation tank by 10% inoculum concentration.The culture medium of seed and fermentation liquid is formed (percentage by weight): molasses 10-12%, H
3PO
40.25-0.3%, (NH
4)
2SO
40.5%, carbamide 0.3%, MgSO
40.03%, ZnSO
40.006%, surplus is a water, pH5.5.Sterilized 20 minutes for 121 ℃.
Fermentation parameter: tank pressure 0.05mPa, 120 rev/mins of mixing speeds, 30 ℃ of temperature, pH5-7 ferments and adds the hydrogen peroxide of fermentation liquid total amount (weight) 0.01% after 8 hours, continues fermentation 4 hours.
(1.2) fermentation ends is 5 * 10 with fermentation liquid dosage
3After the ultraviolet of erg/milliliter shines, 4000 rev/mins of centrifugal collection yeast pastes, yeast paste with aqueous isopropanol in yeast paste: the ratio (weight ratio) of isopropyl alcohol=1: 9 is made into yeast milk, handled two hours, vacuum filtration, yeast slag thalline.In thalline: buffer=ratio (weight ratio) of 1: 3 adds the phosphate buffer of pH8.2, and the amount of press reaction-ure mixture (weight) 0.01% simultaneously adds lysozyme, and room temperature was handled after 20 hours, 5000 rev/mins centrifugal must supernatant.
(1.3) the supernatant cryogenic vacuum concentrates, and lyophilization again gets the yeast multienzyme cluster lyophilized powder.
(2) preparation of liposome
(2.1) 5 kilograms of saturated soybean lecithins, 4 kilograms in cholesterol, double hexadecyl acid ester are dissolved in 80 kilograms of ether for 0.5 kilogram, are the film material behind the mix homogeneously;
(2.2) in order with 4 kilograms of mannitol micropowders, 0.1 kilogram of EDTA disodium salt, join in the film material, decompression stirs;
(2.3) add 80 kilograms of yeast multienzyme cluster lyophilized powders, stir 50 rev/mins of speed;
(2.4) remove ether at 20-35 ℃ of rotary evaporation, obtain having the liposome powder body of mobile yeast multienzyme cluster.
(3) microencapsulation
(3.1) contain the monomeric acetone soln of 50% methylsiloxane and 10 kilograms of alcoholic solution mixing wall-forming material film forming solutions to 6 kilograms;
(3.2) the liposome powder suspension of 100 kilograms of yeast multienzyme clusters on fluid bed, temperature of charge: 40-50 ℃;
Form with spraying in (3.3) 15 minutes sprays into wall material solution in the fluid bed, drying and forming-film under 70 ℃ temperature,
(3.4) 100 mesh sieves are crossed in cooling, get the yeast multienzyme cluster powder product of microencapsulation.
Embodiment two
(1) preparation of yeast multienzyme cluster:
Bakery yeast Saccharomyces cerevisiae is activated to three grades of kinds, inserts fermentation tank by 10% inoculum concentration.The culture medium of seed and fermentation liquid is formed (percentage by weight): molasses 10-12%, H
3PO
40.25-0.3%, (NH
4)
2SO
40.5%, carbamide 0.3%, MgSO
40.039%, ZnSO
40.006%, surplus is a water, pH5.5.Sterilized 20 minutes for 121 ℃;
Fermentation parameter: tank pressure 0.01mpa, 150 rev/mins of mixing speeds, 25 ℃ of temperature, pH5-7,4 hours afterfermentation liquid total amount (weight) H of 0.001% ferment
2O
2, continue fermentation 18 hours;
Fermentation ends is 5 * 10 with fermentation liquid dosage
4Behind the ultraviolet radiation of erg/milliliter, 4000 rev/mins of centrifugal collection yeast pastes, yeast paste with aqueous isopropanol in yeast paste: the ratio (weight ratio) of isopropyl alcohol=9: 1 is made into yeast milk, handled two hours, vacuum filtration, yeast slag thalline.In thalline: the ratio of buffer=1: 10 (weight ratio) adds the phosphate buffer of pH8.2, and the amount of press reaction-ure mixture (weight) 0.01% simultaneously adds lysozyme, and room temperature was handled after 20 hours, 5000 rev/mins centrifugal must supernatant.The supernatant cryogenic vacuum concentrates, and lyophilization again gets the yeast multienzyme cluster lyophilized powder.
(2) preparation of liposome:
10 kilograms of saturated soybean lecithins, 1 kilogram in cholesterol, double hexadecyl acid ester are dissolved in 80 kilograms of ether for 0.1 kilogram, are the film material behind the mix homogeneously;
With 1 kilogram of mannitol micropowder, 0.5 kilogram of EDTA disodium salt, join in the film material in order, decompression stirs;
Add 80 kilograms of yeast multienzyme cluster lyophilized powders, stir 50 rev/mins of speed;
Remove ether at 20-35 ℃ of rotary evaporation, obtain having the liposome powder body of mobile yeast multienzyme cluster.
(3) microencapsulation
Contain 50% the monomeric acetone soln of methylsiloxane and 10 kilograms of alcoholic solution mixing wall-forming material film forming solutions with 6 kilograms;
With the liposome powder suspension of 100 kilograms of yeast multienzyme clusters on fluid bed, temperature of charge: 40-50 ℃;
Form with internal spraying sprayed into wall material film forming solution in the fluid bed in 15 minutes; Drying and forming-film under 70 ℃ temperature;
100 mesh sieves are crossed in cooling, get the yeast multienzyme cluster powder product of microencapsulation.
Embodiment three
(1) preparation of yeast multienzyme cluster
Bakery yeast Saccharomyces cerevisiae is activated to three grades of kinds, inserts fermentation tank by 10% inoculum concentration.The culture medium of seed and fermentation liquid is formed (percentage by weight): molasses 10-12%, H
3PO
40.25-0.3%, (NH
4)
2SO
40.5%, carbamide 0.3%, MgSO
40.03%, ZnSO
40.006%, surplus is a water, pH5.5.Sterilized 20 minutes for 121 ℃;
Fermentation parameter: tank pressure 0.08mPa, 100 rev/mins of mixing speeds, 35 ℃ of temperature, pH5-7,6 hours afterfermentation liquid total amount (weight) H of 0.005% ferment
2O
2, continue fermentation 10 hours;
Fermentation ends is 5 * 10 with fermentation liquid dosage
5The ultraviolet radiation of erg/milliliter, 4000 rev/mins of centrifugal collection yeast pastes, yeast paste with aqueous isopropanol in yeast paste: the ratio (weight ratio) of isopropyl alcohol=1: 20 is made into yeast milk, handled two hours, vacuum filtration, yeast slag thalline.In thalline: buffer=ratio (weight ratio) of 1: 3 adds the phosphate buffer of pH8.2, and the amount of press reaction-ure mixture (weight) 0.01% simultaneously adds lysozyme room temperature to be handled after 20 hours, 5000 rev/mins centrifugal must supernatant.The supernatant cryogenic vacuum concentrates, and lyophilization again gets the yeast multienzyme cluster lyophilized powder.
(2) preparation of liposome
1 kilogram of saturated soybean lecithin, 10 kilograms in cholesterol, double hexadecyl acid ester are dissolved in 80 kilograms of ether for 2 kilograms, are the film material behind the mix homogeneously;
With 5 kilograms of mannitol micropowders, 0.2 kilogram of EDTA disodium salt, join in the film material in order, decompression stirs;
Add 80 kilograms of yeast multienzyme cluster lyophilized powders, stir 50 rev/mins of speed;
Remove ether at 20-35 ℃ of rotary evaporation; Obtain having the liposome powder body of mobile yeast multienzyme cluster.
(3) microencapsulation
With 0.3 kilogram of ethyl cellulose, 0.5 kilogram of PEG-6000,0.3 kilogram of oiliness colorant, be dissolved in the ethanol, mix homogeneously gets 10 kilograms of wall material film forming solutions;
With the liposome powder suspension of 100 kilograms of yeast multienzyme clusters on fluid bed, temperature of charge: 40-50 ℃;
Form with internal spraying sprayed into wall material film forming solution in the fluid bed in 15 minutes; Drying and forming-film under 70 ℃ temperature is crossed 100 mesh sieves, gets the yeast multienzyme cluster powder product of microencapsulation.
Claims (9)
1. the preparation method of the yeast multienzyme cluster of a microencapsulation is characterized in that technological process comprises:
(1) preparation yeast multienzyme cluster:
(1.1) strain fermentation:
Strain: adopt bakery yeast [Saccharomyces cerevisiae];
Seed and fermentation medium: molasses fermented culture fluid, high temperature sterilize;
Sweat: by three grades of seed fermentations and be extended to fermentation tank;
Fermentation condition: 25～35 ℃ of temperature, ventilation 0.6: 1～1: 1,20～300 rev/mins of mixing speeds, tank pressure 0.01～0.08MPa, pH keeps 4.0～7.0, ferments to add the hydrogen peroxide of fermentation liquid total amount (weight) 0.001%～0.01% after 4～8 hours, continues fermentation 4～18 hours;
(1.2) after the fermentation ends, centrifugal collection thalline gets yeast paste;
(1.3) yeast paste of gained is handled with isopropyl alcohol, yeast paste: isopropyl alcohol=1: 0.1～20 (weight ratio), filter, get the yeast slag, the phosphate buffer that adds pH5.5～9.0 again, the yeast slag: buffer=1: 0.1～10 (weight ratio) adds lysozyme room temperature simultaneously and handles the centrifuging and taking supernatant;
(1.4) the supernatant cryogenic vacuum with gained concentrates, and promptly gets yeast multienzyme cluster through lyophilization again;
(2) liposome of preparation yeast multienzyme cluster:
(2.1) saturated soybean lecithin, cholesterol, double hexadecyl acid ester are dissolved in ether and make the film material;
(2.2) in order with the mannitol micropowder, the EDTA disodium salt joins in the film material, and decompression stirs;
(2.3) add the yeast multienzyme cluster micropowder, stir;
(2.4) rotary evaporation is removed ether under 20～35 ℃ of temperature, obtains having the liposome powder body of mobile yeast multienzyme cluster;
Each material mixture ratio (percentage by weight) of above-mentioned preparation liposome is: yeast multienzyme cluster 10-80%, saturated soybean lecithin 1-10%, cholesterol 1-10%, EDTA disodium 0.1-0.5%, mannitol 1-5%, double hexadecyl acid ester 0.1-2%;
(3) microencapsulation:
(3.1) with the high molecular polymer be the wall material, the wall material added in the ethanol that mix homogeneously is a wall-forming material film forming solution;
(3.2) with the liposome powder suspension of capsule heartwood material-yeast multienzyme cluster on fluid bed, the control temperature is at 40～50 ℃;
(3.3) with centrifugal spray wall material film forming solution is sprayed in the fluid bed, make dry materials and film forming;
(3.4) sieve classification is crossed in cooling, promptly gets the product of different size.
2. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1, it is characterized in that step (1.2) fermentation ends in the described technological process after, centrifugal again collection thalline after fermentation liquid shone with ultraviolet, yeast paste.
3. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 2 is characterized in that described ultraviolet irradiating dose is 1 * 10
3～1 * 10
6Erg/milliliter.
4. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1, it is characterized in that in the step (1.3) in the described technological process, handling yeast paste with isopropyl alcohol, is in yeast paste: the ratio of isopropyl alcohol=1: 1～10 (weight ratio) adds isopropyl alcohol in yeast paste.
5. according to the preparation method of the yeast multienzyme cluster of claim 1 or 4 described microencapsulation, it is characterized in that the time that yeast paste is handled with isopropyl alcohol in the step (1.3) in the described technological process is 30 minutes to 120 minutes.
6. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1, it is characterized in that in the step (1.3) in the described technological process in the yeast slag: the ratio of buffer=1: 1～5 (weight ratio) adds the phosphate buffer of pH7.5～8.5.
7. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1 is characterized in that the amount by 0.1～0.01% (percentage by weight) adds the processing of lysozyme room temperature in the step (1.3) in the described technological process.
8. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1 is characterized in that in the step (1.3) in the described technological process, and the time of handling the yeast slag with said phosphate buffer and lysozyme is 10 to 20 hours.
9. the preparation method of the yeast multienzyme cluster of microencapsulation according to claim 1, it is characterized in that in the described encapsulation process step that described high molecular polymer wall material is one or more the compositions in ethyl cellulose, PEG-6000, the methylsiloxane monomer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB01127736XA CN1136834C (en) | 2001-08-14 | 2001-08-14 | Method for preparing microencapsulated yeast multienzyme cluster |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB01127736XA CN1136834C (en) | 2001-08-14 | 2001-08-14 | Method for preparing microencapsulated yeast multienzyme cluster |
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|---|---|
| CN1401314A CN1401314A (en) | 2003-03-12 |
| CN1136834C true CN1136834C (en) | 2004-02-04 |
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ID=4667653
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|---|---|---|---|
| CNB01127736XA Expired - Fee Related CN1136834C (en) | 2001-08-14 | 2001-08-14 | Method for preparing microencapsulated yeast multienzyme cluster |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100389754C (en) * | 2005-10-25 | 2008-05-28 | 天津大学 | Microcapsule preparing process utilizing waste beer yeast cell as wall material |
| CN104195180B (en) * | 2014-08-08 | 2017-08-01 | 广西湘桂酵母科技有限公司 | The method of fermenting and producing high density yeast extract emulsion |
| CN109568242A (en) * | 2019-01-23 | 2019-04-05 | 上海铮信生物科技有限公司 | Phosphatide wraps up composite yeast PCY and the preparation method and application thereof |
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2001
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