CN100544815C - A kind of method of utilizing the microcapsule wall material of yeast cell to prepare safety non-toxic good property - Google Patents
A kind of method of utilizing the microcapsule wall material of yeast cell to prepare safety non-toxic good property Download PDFInfo
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Abstract
The present invention relates to a kind of method of utilizing the microcapsule wall material of yeast cell to prepare safety non-toxic good property, relate to the cultivation of yeast cells, the method for processing, further relate to by this method prepared microcapsule wall material and bio-microcapsule.The yeast cells that to cultivate under appropriate condition directly or again is suspended in Tween-80 or TritonX-100, cetrimonium bromide, lauryl sodium sulfate, ethyl acetate, ethanol, hydrochloric acid, NaOH, potassium chloride and sodium chloride etc. to have self-dissolving and promotes in the autolysis promoter of function, carrying out oscillation treatment under 40～60 ℃ the temperature after 24～48 hours, centrifugal, washing, freeze-drying promptly obtains described microcapsule wall material.Yeast microcapsule wall material and active material are carried out that high frequency contacts etc. and just can make size evenly, not cohering, and the surface does not have the not bio-microcapsule of embedding core material.By yeast cells being carried out reached increase core material embedding rate after modification is handled, and can be used for the purpose of water-soluble substances microencapsulation.
Description
Technical field
The present invention relates to a kind of method of utilizing the microcapsule wall material of yeast cell to prepare safety non-toxic good property, relate to the cultivation of yeast cells, the method for processing, further relate to by this method prepared microcapsule wall material and bio-microcapsule.
Technical background
Microcapsules technology claims microencapsulation again, is one of the new and high technology of 21 century industrial circle focus development such as food, medicine.Microcapsules are made up of core and wall material, have the protection material and avoid the influence of environment, reduce toxicity, shelter disagreeable taste, and the control core discharges, and extends storage period, and it is easy to carry and transport to change states of matter, change rerum natura make can not be compatible the even function such as mixing of composition.These functions make microcapsules technology become effective commercialization method in the industrial circle, have purposes widely in fields such as medicine, spices, food industry.By to the active material of sensitivity or the microencapsulation of volatile component, their timings, target are discharged, not only strengthened its service efficiency, expanded range of application, guaranteed its optimal dose, greatly strengthened their cost performance, and made microencapsulation technology no longer be confined to improve added value of product, but become a kind of brand-new can give new functional components do not have with can be than the technology of superiority.
As the new technology that can improve the functional materials performance, since the NRC company of the U.S. in 1954 utilizes microcapsules technology to be developed into first generation NCR microcapsules and puts on market, microcapsules technology has obtained the development of advancing by leaps and bounds, and the publication of annual relevant microcapsules all increases with the index multiple.Microcapsules technology develops rapidly abroad at present, and only 2002, the patent of relevant microcapsules just had 1000 remainders.But because cost is too high, be difficult to amplify, perhaps make numerous microencapsulation patents and method can't realize industrialization owing to range of application is too narrow.At present, the food of microencapsulation and medicine have occupied grease and the spices and essence in the sizable share, particularly food abroad in its total amount, and the share of microencapsulation is near 20%～35%, and the trend that constantly increases in addition.In recent years, there had been many development in China in the application facet of microcapsules technology, but with abroad comparing, domestic microcapsules technology still is in the starting stage, and the import microcapsules still play leading role aborning.
The selection of wall material is the key of the good microcapsule product of processability.The microcapsule wall material that uses mainly contains natural, semi-synthetic and the synthetic high polymer three major types at present.Natural macromolecular material comprises protein-based (as gelatin, gelatine, fibrin, hemoglobin etc.), natural plant gum class (as natural gum, agar, sodium alginate deer horn glue etc.), wax class (as paraffin, rosin, beeswax etc.), alginates and shitosan class etc.Be characterized in nontoxic, good film-forming property, good stability, but bad mechanical strength, raw material quality instability.Semi-synthetic macromolecular material mainly is a cellulose family, as CMC, and cellulose acetate, cellulose acetate phthalate ester etc.Be characterized in that toxicity is little, viscosity is big, and solubility increases behind the salify, but non-refractory, acid resistance is poor, facile hydrolysis.Synthesized polymer material has the homopolymers class, the condensation polymer class, and copolymer analog, as acrylic resin, epoxy resin, polyvinyl lactam, poly-third ethene etc.Wherein, the most frequently used synthetic material is polylysine (PLL), though good chemical stability and film forming are arranged, can cause inflammatory reaction in human body, is difficult to biodegradation.In addition, liposome is the another approach of protection sensitiveness material, utilize liposome embedded medicine to obtain using widely, but the use of expensive lecithin and organic solvent has limited its application aspect the food micro capsule industrialization.Along with the rise of uphold nature tide, it is a kind of inevitable to use natural macromolecular material to become as microencapsulation wall material.Though new Microencapsulation Method and technology constantly emerge, they are the wall material with protein, carbohydrate and wax mostly, the microcapsules fragility of gained, little to core material adaptability, not withstand voltage, thermo-labile, poor processability, and the granular size of product is inhomogeneous, therefore, the but also few of suitability for industrialized production can be used for, also poisonous solvent or auxiliary material etc. can be used in the process of the microencapsulation that has.For this reason, solving preparation cost safety issue too high and wall material and auxiliary material is one of urgent problem in the microcapsules research, seek raw material be easy to get, inexpensive, applied widely, the microencapsulation preparation is simple, will greatly promote the development of microcapsules technology to the wall material of the mankind and ecological environment security.
The yeast cells safety non-toxic, size is even, and natural eucaryotic cell structure makes it have the potential of embedding substance.Good dispersiveness is arranged in water, and conforming property is strong, and not only material is easy to get, and its look shallow, mouthfeel is soft, through suitable modification, can become a kind of novel, desirable microcapsule wall material fully.Than existing microencapsulation technology, the yeast microcapsules have unique advantage:
(1) the okioplast wall and the internal layer cell membrane of yeast cells carbohydrate composition have constituted natural bilayer vesicles cavity configuration, can avoid the volatilization of volatile materials, also can avoid illumination, the caused oxidation deterioration of oxygen;
(2) need not add any additives in the microencapsulation process, only need water, yeast cells to contact with the high frequency of active material and get final product;
(3) core material discharges easily: as long as run into moist mucous membrane such as tongue or schneiderian membrane, need not broken wall just active material can discharge.The bioadhesive that yeast cells is natural discharges active matter mass-energy for a long time;
(4) non-thermoelastic sack cavity structure makes the parcel core material wherein can be because of not being subjected to hot-extrudable or because of frying in shallow oil, bake, explode, boiling and lose in the food processing process; In fact, yeast cell wall has toughness and pliability, and the beta glucan of supportint cell wall physical strength is embarrassed decomposed substance, therefore, handles even the yeast microcapsules pass through the pressure cooker pressurized, heated or freeze, and core material can not leak yet.
(5) yeast cells is easy to cultivate, and is safe, nontoxic, made it become the most economic microcapsule wall material.
In addition, the cell wall constituent mannosan and the glucan of yeast are bioactivator, and glucan has the generation, the macrophage in the activation animal body that stimulate animal body endolymph cell, lures the effect that animal produces nospecific immunity, improves survival rate candida albicans disease into.And manna oligosacchride (being called for short MOS) is closely similar with the acceptor of pathogen on the intestines wall, with class fourth matter very strong binding ability is arranged, can stop pathogen to be adsorbed at the animal intestine cell surface, add the quantity that manna oligosacchride can reduce enteric pathogenic bacteria in feed, some report claims that MOS is " a pathogen adsorbent " or " pathogen scavenger ".Therefore, yeast cells is a kind of desirable microcapsule wall material.The Fluid Technologies Plc. company that set up in 2000 produces the yeast microcapsules of flavor substance specially.Japan has also succeeded in developing the yeast microcapsule product of grease.
It is fat-soluble requiring core material on the yeast cells microcapsules principle, and that fat-soluble core material enters the complexity of yeast cells is different, reported yeast microcapsule method is limited to the microencapsulation of essential oil class flavor substance more, bibliographical information is arranged recently, and the yeast cells recombinant can be used as the transport agent of oral drugs., handle by yeast cells being carried out modification, can reach increases the core material embedding rate, and can be used for the purpose of water-soluble substances microencapsulation for this reason.
Summary of the invention
The objective of the invention is to handle, a kind of method for preparing the microcapsule wall material and the bio-microcapsule of safety non-toxic good property is provided by yeast cells being carried out modification.This method can reach increases the core material embedding rate, and can be used for the purpose of water-soluble substances microencapsulation.
Therefore, first purpose of the present invention provides a kind of method for preparing the microcapsule wall material of safety non-toxic good property, comprising:
(1) adopting yeast cells is starting strain, after inclined-plane cultivation and seed culture, is seeded in and shakes the bottle cultivation in the fermentation medium;
(2) with the cell centrifugation of gained, after the washing, freeze-drying or be suspended in again and have self-dissolving and promote carrying out oscillation treatment under 40～60 ℃ the temperature after 24～48 hours in the autolysis promoter of function, centrifugal, washing, freeze-drying promptly obtains described microcapsule wall material.Arbitrary Tween-80 or TritonX-100, cetrimonium bromide, lauryl sodium sulfate, ethyl acetate, ethanol, hydrochloric acid, NaOH, potassium chloride and the sodium chloride of being selected from of wherein said autolysis promoter;
The effect that is noted that the autolysis promoter that is added is to promote aqtocytolysis, and if added the autolysis promoter of high concentration the reaction time shorter, if added the autolysis promoter of low concentration the reaction time longer.Therefore in the suitable concn scope, add the autolysis promoter of variable concentrations, those skilled in the art obviously can produce a desired effect by control time length.
In another embodiment, the adding weight of described Tween-80 or TritonX-100 is 0.1～5%; Or
The adding weight of described cetrimonium bromide is 0.1～5%; Or
The adding weight of described lauryl sodium sulfate is 0.1～5%; Or
The adding weight of described ethyl acetate is 0.5～10%; Or
The adding weight of described ethanol is 0.5～10%; Or
The adding weight of described hydrochloric acid is 0.5～10%; Or
The adding weight of described NaOH is 1～20%; Or
The adding weight of described sodium chloride is 1～20%; Or
The adding weight of described potassium chloride is 1～20%;
In another embodiment, the step of shaking the bottle cultivation is as follows: the inclined-plane seed activation inserted seed culture medium after 4 hours, cultivate after 24 hours again and to insert the 250mL that the 50mL fermentation medium is housed by 10% inoculum concentration and shake and carry out fermented and cultured in the bottle, fermentation time 24 hours, fermentation temperature 28-32 ℃, rotating speed 180r/min.
Also in another embodiment, the step that yeast cells is handled is as follows: the yeast cells of cultivation is centrifugal, after the washing, directly freeze drying, or be suspended in again in 0.1～20% the autolysis promoter solution, after handling 24～48 hours under 40～60 ℃, centrifugal, freeze drying promptly obtains described microcapsule wall material.
Second purpose of the present invention provides the method for utilizing prepared microcapsule wall material to prepare bio-microcapsule, comprising:
Step (1)-(2), as hereinbefore;
(3) the yeast microcapsule wall material after the freeze-drying is carried out microencapsulation and handle, as contacting with the high frequency of active material etc.
(4) if desired, adopt the content of core material in the high effective liquid chromatography for measuring microcapsules.
Wherein, embedding degree definition (down together): embedding degree (%)=actual embedding amount/microcapsules weight * 100%
High frequency contact definition (down together): with the frequency that can realize that the fully mixed rotating speed of solution component vibrates.Rotating speed 50rpm/min well known in the art is above or more than the pressurization 2MPa, for example the above vibration frequency of 50-75rpm/min, 75-200rpm/min or 200rpm/min.
In a specific embodiments, the step of utilizing described wall material to prepare microcapsules is as follows: the 1g yeast cells is contacted with the high frequency of antioxidant solution of 1-20mL1% carry out embedding.Take out after 24 hours, the not antioxidant of embedding of surface, freeze drying are removed in centrifugal, washing.
In another specific embodiments, described antioxidant is 1% chlorogenic acid, and described solution is the aqueous solution.The amount of 1% chlorogenic acid that is added in another embodiment, is 1-20mL.
In another embodiment, described antioxidant is 1% resveratrol, and described solution is ethanolic solution.
After the microcapsule wall material modification is handled, the wall material of yeast cells preparation all has in various degree raising to the embedding degree of water-soluble chlorogenic acid and fat-soluble resveratrol, embedding degree to chlorogenic acid does not wait than having improved 3.75%～43.52% before the modification respectively, and the embedding degree of resveratrol has then improved 1.28%～25.06% not to be waited.
The 3rd purpose of the present invention provides the condition of culture that is suitable for cultivating above-mentioned yeast strain.
In a specific embodiments, described bacterial strain is saccharomyces cerevisiae (Saccharomyces cerevisiae Han.), slant medium is the PDA culture medium, seed culture medium and fermentation medium are high glucose medium, consist of: sucrose, YE, inorganic nitrogen-sourced employing center combination design optimization, replenish necessary mineral matter again.
In another embodiment, described seed culture medium and fermentation medium are the YPD culture medium, and be composed as follows: peptone 20.0g, yeast extract 10.0g, glucose 10.0g, add water to 1000mL, pH4-5.
In another embodiment, described seed culture medium and fermentation medium are and produce the fat culture medium, and be composed as follows: peptone 3.0g, yeast extract 8.0g, sucrose 170.0g, add water to 1000mL.
Also in another embodiment, seed culture medium and fermentation medium all are bean sprouts juice SM, are formulated as follows: get moyashi 200.0g, clean, put into water and boil 30min, filtered through gauze.Get bean sprouts juice with sucrose 30.0g, add water, regulate pH=7.2 to 1000mL.
Under the same conditions, in four kinds of culture mediums, the prepared wall material of the yeast cells that utilizes high glucose medium to cultivate is the highest to the embedding degree of chlorogenic acid and resveratrol, be respectively: 8.76% and 4.81%, higher by 10.2% respectively than three kinds of conventional Yeast Cultivation bases, 12.6%, 26.4% and 12.9%, 32.9%, 17.6%.
The 4th purpose of the present invention provides the microcapsule wall material by the prepared safety non-toxic good property of said method.
The 5th purpose of the present invention provides by the prepared bio-microcapsule of said method.
Beneficial effect of the present invention: the yeast cells safety non-toxic, be easy to cultivate, natural eucaryotic cell structure makes its potential with embedding substance, through suitable modification, has become a kind of novel, desirable microencapsulation wall material.According to the present situation of the market demand and China, carry out the research of microcapsules technology at home, particularly study function admirable, cheap and easy to get, environment amenable yeast cells wall material as microencapsulation, will be significant to the development of China's food and pharmaceutical industries.This patent adopts the method for chemical modification, yeast cells is carried out suitable processing, reached increase core material embedding rate, and can be used for the purpose of water-soluble substances microencapsulation, gained bio-microcapsule size evenly, do not cohere, the surface does not have the core material of not embedding, and the microcapsule formulation that initiative China is had independent intellectual property right has crucial meaning.No matter from theory still from angle of practice, utilize the wall material of yeast cells as microencapsulation, can not only fill up the blank of China in this field, China's medical industry, clinical medicine and food industry are all had important social benefit and economic benefit.
The specific embodiment
The preparation method of embodiment 1 described in to specifications is seeded on the PDA inclined-plane seed culture medium activation with the saccharomyces cerevisiae bacterial classification and inserts high sugared seed culture medium after 4 hours and cultivated 24 hours.By in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle again, fermentation temperature 28-32 ℃, and fermentation time 24 hours, shaking speed 180r/min.Centrifugal, the washing after, freeze drying.
The preparation method of embodiment 2 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts the bean sprouts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access bean sprouts culture medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.Centrifugal, the washing after, freeze drying.
The preparation method of embodiment 3 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts the YPD seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access YPD culture medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.Centrifugal, the washing after, freeze drying.
The preparation method of embodiment 4 described in to specifications, with the saccharomyces cerevisiae bacterial classification be seeded on the PDA inclined-plane seed culture medium activation insert after 4 hours produce the fat seed culture medium and cultivate 24 hours after, produce in the fat culture medium by 10% inoculum concentration access again, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.Centrifugal, the washing after, freeze drying.
The preparation method of embodiment 5 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.1～5% the TritonX-100 solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual TritonX-100 after, freeze drying.
The preparation method of embodiment 6 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.1～5% the Tween-80 solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual Tween-80 after, freeze drying.
The preparation method of embodiment 7 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.1～5% the cetrimonium bromide solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual cetrimonium bromide after, freeze drying.
The preparation method of embodiment 8 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in the solution of 0.1～5% lauryl sodium sulfate, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual lauryl sodium sulfate after, freeze drying.
The preparation method of embodiment 9 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.5～10% the ethyl acetate solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual ethyl acetate after, freeze drying.
The preparation method of embodiment 10 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.5～10% the ethanolic solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual ethanol after, freeze drying.
The preparation method of embodiment 11 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 1～20% the sodium chloride solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual sodium chloride after, freeze drying.
The preparation method of embodiment 12 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 1～20% the Klorvess Liquid, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual potassium chloride after, freeze drying.
The preparation method of embodiment 13 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 1～20% the sodium hydroxide solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual NaOH after, freeze drying.
The preparation method of embodiment 14 described in to specifications, after the saccharomyces cerevisiae bacterial classification is seeded on the PDA inclined-plane seed culture medium activation and inserts seed culture medium after 4 hours and cultivate 24 hours, again by in 10% the inoculum concentration access high glucose medium, liquid amount is that 50mL fermentation culture/250mL shakes bottle, fermentation temperature 28-32 ℃, fermentation time 24 hours, shaking speed 180r/min.After centrifugal, the washing, be suspended in again in 0.5～10% the hydrochloric acid solution, handled 24～48 hours down at 40～60 ℃.Centrifugal then, wash residual hydrochloric acid after, freeze drying.
From the result of the foregoing description, we find by analysis:
(1) the chlorogenic acid yeast microcapsules that make are at 25 ℃/75%RH, and 25 ℃/90%RH and 60 ℃ of following storages are after 10 days, and the retention of chlorogenic acid is respectively (99.00 ± 0.60) %, (99.57 ± 0.88) % and (98.72 ± 0.41) %.In SGF, the release rate of chlorogenic acid in 2h reaches more than 95% in the chlorogenic acid yeast microcapsules, and behind the 5h, chlorogenic acid has discharged fully.Illustrate that yeast cells can make chlorogenic acid avoid the external environment influence, reached the purpose of stabilisation, also obviously do not reduce the rate of release of chlorogenic acid simultaneously.
(2) chlorogenic acid yeast microcapsules are to the removing energy force rate predicted value height of DPPH and hydroxy radical, when chlorogenic acid concentration is 17.54mg/L in the microcapsules ethanol extract, it is higher by 60.57% than predicted value to the hydroxy radical clearance rate, and the clearance rate of DPPH free radical has been improved 9.05%.As seen, after the chlorogenic acid microencapsulation, its removing ability to hydroxy radical and DPPH free radical all is significantly improved.
(3) storage is after 10 days down at 60 ℃ for the resveratrol microcapsules that make, and the resveratrol retention is (99.39 ± 0.37) %; Resveratrol retention after (ca.300 lx) under 25 ℃/75%RH and the 25 ℃/90%RH illumination stores 10 days compares 79.88% and 77.77% of not microencapsulation and has improved 22.16% and 19.75% respectively.
(4) resveratrol water-soluble higher more than 2 times than crystal type resveratrol after the microencapsulation also will exceed more than 1 times than amorphous resveratrol.Show after the resveratrol yeast microencapsulation that therefore its bioavilability has also improved 1～2 times.
(5) resveratrol yeast microcapsules can force rate predicted value height to the removing of DPPH and hydroxy radical.When resveratrol concentration in the microcapsules ethanol extract is 9.64 and during 15.60mg/L, its to the removing ability of hydroxy radical respectively than predicted value high 50.75% and 13.70%.When resveratrol concentration in the microcapsules ethanol extract is respectively 10.18,19.28 and during 31.22mg/L, higher by 28.08% than microencapsulation person not respectively to the clearance rate of DPPH free radical, 22.71% and 18.26%.As seen, after the microencapsulation of resveratrol yeast, its removing ability to hydroxy radical and DPPH free radical has all obtained remarkable lifting.
(6) resveratrol of yeast cells microencapsulation in SGF in the 90min release rate reach 90%, this release characteristics for improving resveratrol because metabolism and to drain the too rapid low bioavilability that causes very favourable.Because resveratrol can't accumulate in EV tissue and blood plasma in the resveratrol half-life extremely short, thereby the sustained release performance of microencapsulation resveratrol is very important for the biologically active of keeping resveratrol.
(7) gained bio-microcapsule size is not evenly cohered, and the surface does not have the core material of not embedding.
Claims (6)
1. a method of utilizing yeast cells to prepare microcapsule wall material is characterized in that, this method comprises the steps:
(1) adopting yeast cells is starting strain, through the inclined-plane cultivate and seed culture after, be seeded in and shake a bottle cultivation in the fermentation medium, centrifugal after, use the distilled water washed cell;
(2) with the cell freeze-drying of gained or be suspended in again and have self-dissolving and promote carrying out oscillation treatment under 40～60 ℃ the temperature after 24～48 hours in the autolysis promoter of function, centrifugal, washing, freeze-drying promptly obtains described microcapsule wall material; Wherein said autolysis promoter is arbitrary to be selected from:
Adding weight is 0.1～5% Tween-80 or TritonX-100; Or
Adding weight is 0.1～5% cetrimonium bromide; Or
Adding weight is 0.1～5% lauryl sodium sulfate; Or
Adding weight is 0.5～10% ethyl acetate; Or
Adding weight is 0.5～10% ethanol; Or
Adding weight is 0.5～10% hydrochloric acid; Or
Adding weight is 1～20% NaOH; Or
Adding weight is 1～20% sodium chloride; Or
Adding weight is 1～20% potassium chloride.
2. the method for claim 1 is characterized in that described bacterial strain is saccharomyces cerevisiae (Saccharomyces cerevisiaeHan.), and slant medium is the PDA culture medium, and seed culture medium is a high glucose medium, and fermentation medium is a high glucose medium.
3 the method for claim 1 is characterized in that:
Seed culture medium and fermentation medium are the YPD culture medium, and be composed as follows: peptone 20.0g, yeast extract 10.0g, glucose 10.0g, add water to 1000mL, pH4-5.
4. the method for claim 1 is characterized in that: seed culture medium and fermentation medium are and produce the fat culture medium, and be composed as follows: peptone 3.0g, yeast extract 8.0g, sucrose 170.0g, add water to 1000mL.
5. the method for claim 1 is characterized in that:
Seed culture medium and fermentation medium all are bean sprouts juice SM, are formulated as follows: get moyashi 200.0g, clean, put into water and boil 30min, use filtered through gauze, get bean sprouts juice with sucrose 30.0g, add water to 1000mL, regulate pH=7.2.
6. according to each method prepared microcapsule wall material among the claim 1-5.
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|---|---|---|---|---|
| CN104336574B (en) * | 2014-11-10 | 2019-08-20 | 天津春发生物科技集团有限公司 | A kind of method that biology-frozen drying method prepares yeast microcapsules |
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