CN101250515A - Method for preparing micro-encapsulation glucose oxidase - Google Patents
Method for preparing micro-encapsulation glucose oxidase Download PDFInfo
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- CN101250515A CN101250515A CNA2008100201344A CN200810020134A CN101250515A CN 101250515 A CN101250515 A CN 101250515A CN A2008100201344 A CNA2008100201344 A CN A2008100201344A CN 200810020134 A CN200810020134 A CN 200810020134A CN 101250515 A CN101250515 A CN 101250515A
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Abstract
The invention relates to a method for preparing microencapsulation glucose oxidase, which belongs to the technical field of food processing. The invention aims at improving the quality of flour to researche a method for preparing new flour modifying agent, namely, the microencapsulation glucose oxidase, chitose-calcium alginate-glucose oxidase microcapsule which is obtained through the method is copolymer hydrogel which is obtained through double cross-linking, both amidogen and carboxyl group are fixed, and the whole system is a reticular formation. The method controls the catalytic speed of the microencapsulation glucose oxidase through adjusting the molecular weight and the degree of deacetylation of microcapsule exine chitose, thereby the method is beneficial for oxidation-linking of gluten protein and for increasing the quality of the flour and flour products, simultaneously, the glucose oxidase after microencapsulation avoids the contact between enzyme and the flour and improves storage and the stability, and the microencapsulation glucose oxidase is excellent flour modifying agent and accords with practical production and application. The process of the method is simple, safe and convenient, and accords with the requirements of environmental protection and utility.
Description
Technical field
The present invention relates to the preparation method of micro-encapsulation glucose oxidase, be used to strengthen the biceps of wheat gluten, improve the quality of flour and the finished product thereof, belong to food processing technology field.
Background technology
Whole meal flour is one of main food crop, also be the main raw material of producing food such as bread, steamed bun, noodles, biscuit, cake, so flour industry has consequence in national economy.Along with the raising of people's living standard and the reinforcement of health perception, the flour improver of green non-pollution---zymin (as glucose oxidase) more and more comes into one's own, but various zymins are uneven at the action effect that improves aspect the flower characters in the practical application, and having problem stable, storage property difference, the flour improver of therefore researching and developing high efficiency, practicality, green safety is particularly important.
Glucose oxidase (GOD) is one of zymin at present of greatest concern, has been applied to (Chinese patent 200610037642.4 among the actual production; United States Patent (USP) 20070141201; International monopoly WO 99152372).GOD can generate glucose oxidase gluconic acid and hydrogen peroxide (H
2O
2), the sulfydryl in latter's oxidation mucedin generates disulfide linkage, thereby has improved the weave construction of dough greatly, has finally improved the quality of flour product.But free GOD still has many weak points, and at first GOD belongs to quick oxygenant, and catalysis speed is very fast, has just generated H in the forming process of dough
2O
2, and the H that generates too early
2O
2Can make dough exsiccation hardening, cause the final inferior quality that bakes; In addition, GOD is less stable in the flour medium, Rakotozafy, and reports such as L, GOD has just lost 25% at the preceding 5min that dough forms, and ensuing 20min has 20% enzyme deactivation again, so the action effect of GOD is not ideal enough.
Since Abraham invention APA microcapsule, microcapsulary is widely used in biological technical fields such as the immobilization, medicine sustained release of enzyme.Poncelet in 1992 has reported that the inner gel of alginate calcium-enzyme absorption-chitosan outside embedding legal system is equipped with the technology of microcapsule, for the immobilization of enzyme has launched more wide prospect.Sodium alginate is the natural polymer that is present in the brown alga, it and polyvalent cation (as calcium ion) instantaneous gelation of meeting when contacting, this gel has porousness and powerful absorption supporting capacity, can utilize this characteristic that materials such as protein, enzyme are adsorbed embedding.This preparation process is simple, and core do not participate in the forming process of microcapsule, has avoided deleterious conditions such as high temperature, organic solvent, is beneficial to maintenance by the activity of embedding thing.Chitosan extensively is present in arthropodan shell and the zootic cell walls; the straight-chain polysaccharide of making through deacetylationization; primary amino on its molecular chain can with the carboxyl generation electrostatic interaction on the sodium alginate molecular chain; thereby at sodium alginate micro gel capsule surface recombination one deck polyelectrolyte film; so both improved the stability and the embedding rate of microcapsule, can regulate by the release rate of embedding thing again.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of micro-encapsulation glucose oxidase, this preparation method adopts the principle of the inner gel of alginate calcium-outside embedding of enzyme absorption-chitosan, and is safe and simple, and has the characteristics of high adsorption rate and high enzyme retention rate alive.
Technical scheme of the present invention is: the preparation method of micro-encapsulation glucose oxidase comprises the steps:
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass concentration that forms transparent and homogeneous is the colloidal sol of 1%-5%; With molecular weight 5,000-450,000, deacetylation is that the chitosan of 70%-98% adds in the glacial acetic acid aqueous solution of mass concentration 0.1%-2% (optimum concn 1%), generating concentration is the transparent chitosan solution (pH3.0-5.0) of 0.5%-2%;
2) preparation contains the vegetable oil solution of 1.0%-3.0%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds Glacial acetic acid and make the mass concentration of Glacial acetic acid reach 0.5%-1.5% in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) in the sodium alginate colloidal sol that step 1) obtains, add CaCO
3, add-on is 0.1-1.0g/100mL, high-speed stirring is mixed, and ultrasonic wave homogeneous 5-10min, vacuum outgas obtains the water mixed solution;
4) prepare acetate-sodium acetate buffer of pH3.0-5.0 (best pH4.0), and prepare the glucose oxidase GOD solution of mass concentration 0.5%-2% with this damping fluid;
5) the water mixed solution that step 3) is obtained is at 800-1, slowly join step 2 under the 500rpm high-speed stirring) in the resulting oil phase mixed liquor I, the part by weight of water mixed solution and oil phase mixed solution is 1/2-1/4, add oil phase mixed liquor I I behind the stirring and emulsifying 1-2min, make final water mixed solution and total oil phase mixed solution part by weight at 1/3-1/6, continue to stir 10-20min, adding distil water is breakdown of emulsion 20min under the 200-400rpm stirring at low speed;
6) the oil-water mixture frozen centrifugation behind the breakdown of emulsion that step 5) is obtained, 2,000-5,000rpm, 10-30min, reject upper strata fluid, lower floor's material washs centrifugal further removal fluid with the Tween 80 of mass concentration 1%-3% (optimum concn 1%), and washings is removed in the redistilled water washing, obtain alginate calcium bead CaAlg, dry matter content 6-10mg/mL;
7) the alginate calcium bead CaAlg equal-volume that GOD solution and the step 6) that step 4) is made obtains mixes, and 4 ℃ adsorb 0.5-3h down, obtain adsorbing the alginate calcium bead CaAlg-GOD of glucose oxidase;
8) CaAlg-GOD that step 7) is made slowly joins in the chitosan solution that step 1) makes, and 800-1,300rpm stir 10-20min fast, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule (CH-CaAlg-GOD) that obtain;
9) prepare respectively that mass concentration is 10%, the oligomeric maltodextrin of DE value 15-20 and mass concentration are 2% beta-cyclodextrin, 60 ℃ are mixed down and obtain helping dried solution at 1: 1;
10). the CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, CH-CaAlg-GOD is 1 with helping the volume ratio of dried solution: 8-1: 12, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 10,000-40,000rpm, feed rate are 10-30mL/min, inlet temperature 170-220 ℃, temperature out 40-70 ℃, make the chitin-alginic acid calcium-glucose oxidase microcapsule of size distribution at 10-100nm.
Chitosan is selected from: four kinds of chitosan molecule amounts are respectively 450,000,200,000,100,000,5,000, corresponding deacetylation is respectively 91%, 87%, 91%, 85%, four kinds of chitosans is added respectively contain in 1% the glacial acetic acid aqueous solution, and making concentration is the transparent chitosan solution of 0.5%-2%.
Beneficial effect of the present invention: chitin-alginic acid calcium-glucose oxidase microcapsule that the present invention adopts aforesaid method to make are through the dual crosslinked gel copolymer that obtains, and amino and carboxyl all is fixed, and whole system is a reticulated structure.The sodium alginate mixed solution forms the emulsification small droplets in oil phase, after adding contains the oil solution of acetate, and the CaCO of drop inside
3Decompose, discharge Ca
2+And CO
2Gas, sodium alginate and Ca
2+Contact instantaneous gelation, generate alginate calcium bead CaAlg, because CO
2Release, CaAlg has porousness and powerful absorption supporting capacity, a large amount of GOD can be adsorbed on bead surface and mesh inside.Near near alginate calcium (pI 3.8) and GOD (pI 4.2) iso-electric point, both institutes are electrically charged minimum, and adsorptive capacity is the highest.Chitosan is an alkaline polysaccharide, has a large amount of positive charges in acid range, can with electronegative alginate calcium rapid polymerization, play parcel and fill out the effect of envelope.
Preparation method's technology of chitin-alginic acid calcium provided by the invention-glucose oxidase microcapsule is simple, safe and effective, meet environmental protection and practical requirement.The release rate of GOD slows down after the micro encapsulation, more helps the formation and the optimization of mucedin, thereby improves the internal structure and the sensible quality of flour products.The catalysis speed of GOD in molecular weight by regulating chitosan and the deacetylation may command microcapsule; thereby reach optimal flour improvement effect; microcapsule wall material has provide protection to the GOD of inside simultaneously; avoided contacting of enzyme and flour; improve stability and the storage property of GOD, be expected to become good flour improver.
Specific embodiments
Embodiment 1: the preparation of chitin-alginic acid calcium-glucose oxidase microcapsule
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass percent that forms transparent and homogeneous is 1.5% colloidal sol; Chitosan (molecular weight 450,000, deacetylation are 91%) adding is contained in the aqueous solution of 1% Glacial acetic acid, and generation concentration is 2.0% hyaline test glycan solution I (pH3.5);
2) preparation contains the vegetable oil solution of 1.0%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds 0.8% Glacial acetic acid in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) get the sodium alginate colloidal sol 30mL that step 1) obtains, add 0.1g crystallite CaCO
3, high-speed stirring is mixed, ultrasonic wave homogeneous 5min, and vacuum outgas obtains the water mixed solution;
4) acetate-sodium acetate buffer of preparation pH4.0, and with glucose oxidase (GOD) solution of damping fluid preparation 0.5%;
5) the water mixed solution that step 3) is obtained is in high-speed stirring (1, slowly join 90mL step 2 000rpm)) in the resulting oil phase mixed liquor I, add 30mL oil phase mixed liquor I I behind the stirring and emulsifying 2min, continue to stir 20min, add 100mL distilled water, 200rpm stirs breakdown of emulsion 20min;
6) 4 ℃ of following frozen centrifugations (3 of the oil-water mixture that step 5) is obtained, 000rpm) 20min, reject upper strata fluid, the centrifugal removal fluid of Tween 80 washings with 1% 2 times, distilled water wash is removed washings 2 times, obtains alginate calcium bead CaAlg (dry matter content 7.7mg/mL);
7) get GOD solution that the 2mL step 4) makes and mix with the alginate calcium bead CaAlg that step 6) with volume obtains, 4 ℃ of absorption 0.5h down obtain adsorbing the alginate calcium bead (CaAlg-GOD) of glucose oxidase;
8) get chitosan solution that the 2mL step 1) makes and slowly join among the CaAlg-GOD that step 7) makes the 10min that vibrates at a high speed, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule (CH-CaAlg-GOD) that obtain.
9). compound concentration is 10% and 2% oligomeric maltodextrin (DE value 15-20) and beta-cyclodextrin respectively, and 60 ℃ of mixing in following 1: 1 obtain helping dried solution;
10). the CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, volume ratio is 1: 8, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 20,000rpm, feed rate are 10ml/min, 180 ℃ of inlet temperatures, 50 ℃ of temperature outs, finally make the glucose oxidase microcapsule of size distribution at 10-80nm.
Embodiment 2: the preparation of chitin-alginic acid calcium-glucose oxidase microcapsule
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass percent that forms transparent and homogeneous is 2.0% colloidal sol; Chitosan (molecular weight 450,000 and 200,000 deacetylations are respectively 91% and 87%) added respectively contain in the aqueous solution of 1% Glacial acetic acid, generate concentration and be all 1.0% hyaline test glycan solution I and II, it is 3.5 and 4.0 in addition that pH divides;
2) preparation contains the vegetable oil solution of 1.5%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds 1.2% Glacial acetic acid in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) get the sodium alginate colloidal sol 30mL that step 1) obtains, add 0.15g crystallite CaCO
3, high-speed stirring is mixed, ultrasonic wave homogeneous 10min, and vacuum outgas obtains the water mixed solution;
4) acetate-sodium acetate buffer of preparation pH4.0, and with glucose oxidase (GOD) solution of damping fluid preparation 1.0%;
5) the water mixed solution that step 3) is obtained is in high-speed stirring (1, slowly join 80mL step 2 200rpm)) in the resulting oil phase mixed liquor I, add 40mL oil phase mixed liquor I I behind the stirring and emulsifying 2min, continue to stir 20min, add 100mL distilled water, 300rpm stirs breakdown of emulsion 20min;
6) 4 ℃ of following frozen centrifugations (4 of the oil-water mixture that step 5) is obtained, 000rpm) 15min, reject upper strata fluid, the centrifugal removal fluid of Tween 80 washings with 1% 2 times, distilled water wash is removed washings 2 times, obtains alginate calcium bead CaAlg (dry matter content 8.9mg/mL);
7) get GOD solution that the 2mL step 4) makes and mix with the alginate calcium bead CaAlg that step 6) with volume obtains, 4 ℃ of absorption 1.0h down obtain adsorbing the alginate calcium bead (CaAlg-GOD) of glucose oxidase;
8) CaAlg-GOD that the 2mL step 7) is made slowly joins among the chitosan solution I that step 1) makes, 10min vibrates at a high speed, add the chitosan solution II that the 2mL step 1) makes again, the 10min that vibrates at a high speed once more, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule (CH-CaAlg-GOD) that obtain.
9). compound concentration is 10% and 2% oligomeric maltodextrin (DE value 15-20) and beta-cyclodextrin respectively, and 60 ℃ of mixing in following 1: 1 obtain helping dried solution;
10). the CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, volume ratio is 1: 10, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 30,000rpm, feed rate are 20ml/min, 190 ℃ of inlet temperatures, 60 ℃ of temperature outs, finally make the glucose oxidase microcapsule of size distribution at 10-100nm.
Embodiment 3: the preparation of chitin-alginic acid calcium-glucose oxidase microcapsule
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass percent that forms transparent and homogeneous is 1.0% colloidal sol; With chitosan (molecular weight 450,000,200,000 and 5,000, deacetylation is respectively 91%, 87% and 85%) add respectively and contain in the aqueous solution of 1% Glacial acetic acid, make concentration and be respectively 0.5%, 1.0%, 1.5% hyaline test glycan solution I, II and III, pH is respectively 3.5,4.0 and 3.8;
2) preparation contains the vegetable oil solution of 1.5%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds 0.5% Glacial acetic acid in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) get the sodium alginate colloidal sol 30mL that step 1) obtains, add 0.1g crystallite CaCO
3, high-speed stirring is mixed, ultrasonic wave homogeneous 10min, and vacuum outgas obtains the water mixed solution;
4) acetate-sodium acetate buffer of preparation pH4.0, and with glucose oxidase (GOD) solution of damping fluid preparation 1.0%;
5) the water mixed solution that step 3) is obtained slowly joins 70mL step 2 under high-speed stirring (900rpm)) in the resulting oil phase mixed liquor I, add 50mL oil phase mixed liquor I I behind the stirring and emulsifying 2min, continue to stir 20min, add 100mL distilled water, 400rpm stirs breakdown of emulsion 20min;
6) 4 ℃ of following frozen centrifugations (5 of the oil-water mixture that step 5) is obtained, 000rpm) 15min, reject upper strata fluid, the centrifugal removal fluid of Tween 80 washings with 1% 2 times, distilled water wash is removed washings 2 times, obtains alginate calcium bead CaAlg (dry matter content 8.7mg/mL);
7) get GOD solution that the 2mL step 4) makes and mix with the alginate calcium bead CaAlg that step 6) with volume obtains, 4 ℃ of absorption 1.5h down obtain adsorbing the alginate calcium bead (CaAlg-GOD) of glucose oxidase;
8) CaAlg-GOD that the 2mL step 7) is made slowly joins among the chitosan solution I that step 1) makes, 10min vibrates at a high speed, add chitosan solution II and III that the 2mL step 1) makes again successively, 10min vibrates at a high speed respectively, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule (CH-CaAlg-GOD) that obtain, spraying drying finally makes the glucose oxidase microcapsule of size distribution at 20-80nm.
9). compound concentration is 10% and 2% oligomeric maltodextrin (DE value 15-20) and beta-cyclodextrin respectively, and 60 ℃ of mixing in following 1: 1 obtain helping dried solution;
10). the CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, volume ratio is 1: 10, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 20,000rpm, feed rate are 15ml/min, 200 ℃ of inlet temperatures, 65 ℃ of temperature outs, finally make the glucose oxidase microcapsule of size distribution at 10-100nm.
Embodiment 4: the preparation of chitin-alginic acid calcium-glucose oxidase microcapsule
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass percent that forms transparent and homogeneous is 1.5% colloidal sol; With chitosan (molecular weight 450,000,200,000,100,000 and 5,000, deacetylation is respectively 91%, 87%, 91% and 85%) add respectively and contain in the aqueous solution of 1% Glacial acetic acid, make concentration and be respectively 0.5%, 1.0%, 2%, 1.5% transparent chitosan solution I, II, III and IV, pH is respectively 3.5,4.0,4.2 and 3.8;
2) preparation contains the vegetable oil solution of 1.0%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds 1.4% Glacial acetic acid in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) get the sodium alginate colloidal sol 30mL that step 1) obtains, add 0.1g crystallite CaCO
3, high-speed stirring is mixed, ultrasonic wave homogeneous 10min, and vacuum outgas obtains the water mixed solution;
4) acetate-sodium acetate buffer of preparation pH4.0, and with glucose oxidase (GOD) solution of damping fluid preparation 1.0%;
5) the water mixed solution that step 3) is obtained is in high-speed stirring (1, slowly join 100mL step 2 300rpm)) in the resulting oil phase mixed liquor I, add 20mL oil phase mixed liquor I I behind the stirring and emulsifying 2min, continue to stir 20min, add 100mL distilled water, 400rpm stirs breakdown of emulsion 20min;
6) 4 ℃ of following frozen centrifugations (5 of the oil-water mixture that step 5) is obtained, 000rpm) 15min, reject upper strata fluid, the centrifugal removal fluid of Tween 80 washings with 1% 2 times, distilled water wash is removed washings 2 times, obtains alginate calcium bead CaAlg (dry matter content 9.4mg/mL);
7) get GOD solution that the 2mL step 4) makes and mix with the alginate calcium bead CaAlg that step 6) with volume obtains, 4 ℃ of absorption 1.0h down obtain adsorbing the alginate calcium bead (CaAlg-GOD) of glucose oxidase;
8) CaAlg-GOD that the 2mL step 7) is made slowly joins among the chitosan solution I that step 1) makes, 10min vibrates at a high speed, add chitosan solution II, III and IV that the 2mL step 1) makes again successively, 10min vibrates at a high speed respectively, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule (CH-CaAlg-GOD) that obtain, spraying drying finally makes the glucose oxidase microcapsule of size distribution at 20-100nm.
9) compound concentration is 10% and 2% oligomeric maltodextrin (DE value 15-20) and beta-cyclodextrin respectively, and 60 ℃ are mixed down and obtain helping dried solution at 1: 1;
10) CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, volume ratio is 1: 12, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 40,000rpm, feed rate are 30ml/min, 210 ℃ of inlet temperatures, 70 ℃ of temperature outs, finally make the glucose oxidase microcapsule of size distribution at 10-100nm.
Claims (2)
1. the preparation method of a micro-encapsulation glucose oxidase is characterized in that it comprises the steps:
1) with sodium alginate at room temperature mechanical stirring it is dissolved in the water, the mass concentration that forms transparent and homogeneous is the colloidal sol of 1%-5%; With molecular weight 5,000-450,000, deacetylation is that the chitosan of 70%-98% adds in the glacial acetic acid aqueous solution of mass concentration 0.1%-2%, generate pH3.0-5.0, concentration is the transparent chitosan solution of 0.5-2%;
2) preparation contains the vegetable oil solution of 1.0%-3.0%span 80, and high-speed stirring and vacuum outgas make the oil phase mixed liquor I, adds Glacial acetic acid and make the mass concentration of Glacial acetic acid reach 0.5%-1.5% in the oil phase mixed liquor I, and thorough mixing gets oil phase mixed liquor I I;
3) in the sodium alginate colloidal sol that step 1) obtains, add CaCO
3, add-on is 0.1-1.0g/100mL, high-speed stirring is mixed, and ultrasonic wave homogeneous 5-10min, vacuum outgas obtains the water mixed solution;
4) prepare acetate-sodium acetate buffer of pH3.0-5.0, and prepare the glucose oxidase GOD solution of mass concentration 0.5%-2% with this damping fluid;
5) the water mixed solution that step 3) is obtained is at 800-1, slowly join step 2 under the 500rpm high-speed stirring) in the resulting oil phase mixed liquor I, the part by weight of water mixed solution and oil phase mixed liquor I is 1/2-1/4, add oil phase mixed liquor I I behind the stirring and emulsifying 1-2min, make final water mixed solution and total oil phase mixed solution part by weight at 1/3-1/6, continue to stir 10-20min, adding distil water is breakdown of emulsion 20min under the 200-400rpm stirring at low speed;
6) the oil-water mixture frozen centrifugation behind the breakdown of emulsion that step 5) is obtained, 2,000-5,000rpm, 10-30min, reject upper strata fluid, lower floor's material washs centrifugal further removal fluid with the Tween 80 of mass concentration 1%-3%, and washings is removed in the redistilled water washing, obtain alginate calcium bead CaAlg, dry matter content 6-10mg/mL;
7) the alginate calcium bead CaAlg equal-volume that GOD solution and the step 6) that step 4) is made obtains mixes, and 4 ℃ adsorb 0.5-3h down, obtain adsorbing the alginate calcium bead CaAlg-GOD of glucose oxidase;
8) CaAlg-GOD that step 7) is made slowly joins in the chitosan solution that step 1) makes, and 800-1,300rpm stir 10-20min fast, the centrifugal chitin-alginic acid calcium-glucose oxidase microcapsule CH-CaAlg-GOD that obtains;
9) prepare respectively that mass concentration is 10%, the oligomeric maltodextrin of DE value 15-20 and mass concentration are 2% beta-cyclodextrin, 60 ℃ are mixed down and obtain helping dried solution at 1: 1;
10). the CH-CaAlg-GOD that step 8) is made joins helping in the dried solution that step 9) makes, CH-CaAlg-GOD is 1 with helping the volume ratio of dried solution: 8-1: 12, mix the back spraying drying, the revolution of centrifugal sprinkling dish is 10,000-40,000rpm, feed rate are 10-30mL/min, inlet temperature 170-220 ℃, temperature out 40-70 ℃, make the chitin-alginic acid calcium-glucose oxidase microcapsule of size distribution at 10-100nm.
2. preparation method according to claim 1, it is characterized in that chitosan is selected from: four kinds of chitosan molecule amounts are respectively 450,000,200,000,100,000,5,000, corresponding deacetylation is respectively 91%, 87%, 91%, 85%, four kinds of chitosans are added respectively contain in 1% the glacial acetic acid aqueous solution, making concentration is the transparent chitosan solution of 0.5%-2%.
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