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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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 239000002502 liposome Substances 0.000 claims abstract description 21
- 102000002568 Multienzyme Complexes Human genes 0.000 claims abstract description 19
- 108010093369 Multienzyme Complexes Proteins 0.000 claims abstract description 19
- 102000004190 Enzymes Human genes 0.000 claims abstract description 7
- 108090000790 Enzymes Proteins 0.000 claims abstract description 7
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 90
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 45
- 238000000855 fermentation Methods 0.000 claims description 41
- 230000004151 fermentation Effects 0.000 claims description 41
- 239000000463 material Substances 0.000 claims description 29
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 20
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 11
- 239000000047 product Substances 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 9
- 239000002893 slag Substances 0.000 claims description 9
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 8
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 8
- 102000016943 Muramidase Human genes 0.000 claims description 8
- 108010014251 Muramidase Proteins 0.000 claims description 8
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 8
- 229960000274 lysozyme Drugs 0.000 claims description 8
- 239000004325 lysozyme Substances 0.000 claims description 8
- 235000010335 lysozyme Nutrition 0.000 claims description 8
- 235000012000 cholesterol Nutrition 0.000 claims description 7
- 235000010355 mannitol Nutrition 0.000 claims description 7
- 239000008363 phosphate buffer Substances 0.000 claims description 7
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 229930195725 Mannitol Natural products 0.000 claims description 6
- 229940088598 enzyme Drugs 0.000 claims description 6
- 239000000594 mannitol Substances 0.000 claims description 6
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical class CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- 235000013379 molasses Nutrition 0.000 claims description 5
- 230000005855 radiation Effects 0.000 claims description 5
- 238000002390 rotary evaporation Methods 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- -1 methyl siloxane Chemical class 0.000 claims description 4
- 229920000642 polymer Polymers 0.000 claims description 4
- 239000007921 spray Substances 0.000 claims description 4
- 239000001856 Ethyl cellulose Substances 0.000 claims description 3
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 claims description 3
- 235000019325 ethyl cellulose Nutrition 0.000 claims description 3
- 229920001249 ethyl cellulose Polymers 0.000 claims description 3
- 239000008118 PEG 6000 Substances 0.000 claims description 2
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000000178 monomer Substances 0.000 claims description 2
- 238000009423 ventilation Methods 0.000 claims description 2
- 239000007853 buffer solution Substances 0.000 claims 2
- RNPXCFINMKSQPQ-UHFFFAOYSA-N dicetyl hydrogen phosphate Chemical compound CCCCCCCCCCCCCCCCOP(O)(=O)OCCCCCCCCCCCCCCCC RNPXCFINMKSQPQ-UHFFFAOYSA-N 0.000 claims 2
- 239000008055 phosphate buffer solution Substances 0.000 claims 2
- 239000010802 sludge Substances 0.000 claims 2
- 241000894006 Bacteria Species 0.000 claims 1
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims 1
- 241001052560 Thallis Species 0.000 claims 1
- 238000005119 centrifugation Methods 0.000 claims 1
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims 1
- MJEMIOXXNCZZFK-UHFFFAOYSA-N ethylone Chemical compound CCNC(C)C(=O)C1=CC=C2OCOC2=C1 MJEMIOXXNCZZFK-UHFFFAOYSA-N 0.000 claims 1
- 230000002934 lysing effect Effects 0.000 claims 1
- 238000012423 maintenance Methods 0.000 claims 1
- 239000012528 membrane Substances 0.000 claims 1
- 238000007873 sieving Methods 0.000 claims 1
- 230000001954 sterilising effect Effects 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 claims 1
- 230000001939 inductive effect Effects 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 239000012530 fluid Substances 0.000 description 8
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 6
- 239000008176 lyophilized powder Substances 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 239000002253 acid Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
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- 150000002148 esters Chemical class 0.000 description 5
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 230000006837 decompression Effects 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 239000004202 carbamide Substances 0.000 description 3
- 235000013877 carbamide Nutrition 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 238000005507 spraying Methods 0.000 description 3
- 238000003828 vacuum filtration Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 229920001503 Glucan Polymers 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 108010012715 Superoxide dismutase Proteins 0.000 description 2
- 230000001476 alcoholic effect Effects 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000005538 encapsulation Methods 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 239000003094 microcapsule Substances 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000003918 blood extract Substances 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
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- 239000003814 drug Substances 0.000 description 1
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- 230000003760 hair shine Effects 0.000 description 1
- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000004223 radioprotective effect Effects 0.000 description 1
- 238000010334 sieve classification Methods 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Manufacturing Of Micro-Capsules (AREA)
Abstract
本发明提供一种适用于护肤品配方的酵母多酶复合体及其微囊化的生物制品—微囊化的酵母多酶复合体的制备方法，其工艺流程包括：发酵—→诱导产酶—→温和提取—→浓缩干燥—→脂质体包埋—→微囊化。本发明工艺设计合理，适合工业化生产；产品性能稳定，适应市场需求。The present invention provides a preparation method of yeast multienzyme complex suitable for formulas of skin care products and its microencapsulated biological product—microencapsulated yeast multienzyme complex. The process includes: fermentation—→inducing enzyme production— →gentle extraction—→concentration and drying—→liposome embedding—→microencapsulation. The invention has reasonable process design and is suitable for industrial production; the product has stable performance and meets market demands.
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)
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 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1401314A CN1401314A (en) | 2003-03-12 |
| CN1136834C true CN1136834C (en) | 2004-02-04 |
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| Application Number | Title | Priority Date | Filing Date |
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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 (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100389754C (en) * | 2005-10-25 | 2008-05-28 | 天津大学 | Method for preparing microcapsules using beer waste yeast cells 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 |
| CN116918909B (en) * | 2023-07-20 | 2025-06-20 | 南通奥凯生物技术开发有限公司 | Acetate microcapsule feed additive and preparation method thereof |
| CN118077890A (en) * | 2023-09-28 | 2024-05-28 | 南昌大学 | An intestinal sustained-release vesicle and microcapsule with taste-masking function and preparation method and application thereof |
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