CN102943069B - Co-immobilization glucose oxidase/catalase microspheres and application thereof in production of gluconic acid or gluconic salt - Google Patents
Co-immobilization glucose oxidase/catalase microspheres and application thereof in production of gluconic acid or gluconic salt Download PDFInfo
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Abstract
The invention discloses co-immobilization glucose oxidase/catalase microspheres and application thereof in preparation of gluconic acid (salt) for catalytic oxidation of glucose. The co-immobilization glucose oxidase/the catalase microspheres can effectively promote covalent linkage of carriers and enzyme molecules, immobilization efficiency is improved greatly, service life of immobilization glucose oxidase (GOD)/catalase (CAT) is prolonged remarkably, the co-immobilization glucose oxidase/the catalase microspheres can be used repeatedly, are low in production cost, and facilitate sustainable development. Gluconic acid (salt) prepared by the co-immobilization glucose oxidase/the catalase microspheres is high in yield, reaction condition is temperate, a device is simple and easy to obtain, and the production method is environment-friendly.
Description
Technical field
The present invention relates to the production method of gluconic acid or its salt, particularly mixed immobilized glucose oxidase/catalase microspheres and produce the application in gluconic acid or its salt at catalytic oxidation of glucose.
Background technology
Gluconic acid is glucose oxidase product, after derivative, can obtain serial gluconic acid product, it comprises calglucon, the products such as Potassium Gluconate, Sunmorl N 60S, Menesia, Zinc Gluconate, Ferrous Gluconate, manganese gluconate, copper gluconate, gluconic acid barium and Gluconolactone.Gluconate series product and Gluconolactone, at home and abroad demand reaches ten thousand tons of 70-80, and output constantly expands.
The production method of calglucon has electrolytic oxidation, bromination method, metal catalytic synthesis method, fermentation method etc.In industrialized production, mainly adopt at present metal catalytic method and fermentation method.
Electrolytic oxidation preparation of gluconic acid is existing suitability for industrialized production abroad, and it is still under test at home, the people such as Shandong in 2000 war light are plated on titanium ruthenium as working electrode, and current efficiency (generating the theoretical consumes power/generation of mole gluconic acid unit of unit mole gluconic acid actual consumption electric weight) can reach 76.50%.
The people such as Wang Zumo and Gao Shutong, with eletrooxidation method, adopts respectively hydrogen peroxide, clorox as oxygenant, and productive rate is respectively 70% and 90%, has realized industrialization pilot scale.But eletrooxidation method needs the strict content of controlling catalyzer effective constituent in reaction solution, and temperature, pH are had to dependency, and intermediate steps is many, and by product is many, and product is difficult to separation, and be difficult to regeneration as the salt of catalyzer, productive rate is lower.Reaction times is longer, and environment is had to larger pollution.
China adopts fermentative Production calglucon more at present, then with the ion-exchange of calglucon process, evaporation concentration, the acid of crystallization synthesis of glucose.Biological fermentation process need to be cultivated many processes such as bacterial classification, strain screening and sterilizing, and comparatively strict to temperature requirement, by product is many, cycle is longer, and in the gluconic acid process of producing because adding the impurity such as thalline, the product purity of affecting glucose acid, thereby its development is badly in need of solving a lot of technical problems.
In recent years, Enzymes Industry is accompanied by the fast development of downstream industry and develops, and in kind and quality, is all greatly improved.Enzyme method technique utilizes zymin directly conversion of glucose to be become to gluconic acid exactly, then passes through the neutralizing effect of alkali, converts it into gluconate series product.Compare with fermentation method, the outstanding feature of enzyme process is exactly not need seed culture, has removed the interference of raw and auxiliary material to reaction system such as microorganism and substratum from, has improved the purity of reaction product, to purifying, brings convenience.There is no seed culture, thereby reaction is more prone to control, produces and also can become more steady.For the producer, enzyme method technique is easy, and equipment is simple, easy to operate, there is no the danger of microbiological contamination, and product is single, and purity is high, is easy to separated and refines, and quality product and yield significantly improve, and product hierarchy reaches food grade and injection stage standard completely.For newly founding the factory, can save seeding tank and extracting section equipment, reduced facility investment.Compare with metal catalytic method, enzyme process also has safe feature.
By being fixed of enzyme, can significantly improve the stability of enzyme.Be not subject to the impact of external environment and inactivation.Immobilized enzyme is easy to and product separation, owing to only having substrate in reaction system, and product,, by immobilized enzyme Separation and Recovery from reaction system, only there is product after reaction finishes in immobilized enzyme, is easy to carry out the refining of product in system, and product purity is high.Immobilized enzyme can repeat to recycle, and greatly reduces the cost of enzyme.
The existing technique of utilizing immobilized enzyme method to prepare gluconate, for example < < chemistry with civilian disclosed method of biotechnology > > the 2nd phase in 2008 " research of Immobilization of Glucose Oxidase by Calcium Alginate Gel Capsules " one is: utilize calcium alginate gel particle fixing glucose oxidase (GOD), the method is easy to operation, reaction conditions is gentle, but this method has only been fixed an enzyme, the H producing in catalytic process
2o
2cannot decompose in time, accelerate the inactivation of GOD and CAT; In addition, immobilized enzyme less stable, reuses number of times few.And for example civilian disclosed method is < < food and pharmaceutical > > the 9th phase in 2011 " immobilized enzyme method is prepared the research of calglucon " one: glucose oxidase and catalase covalency are fixed on the pretreated chitosan of glutaraldehyde, are calglucon by gained immobilized enzyme catalysis conversion of glucose.For another example " a kind of method of preparing mixed immobilized glucose oxidase/catalase microspheres " patent that on September 28th, 2011, disclosed publication number was CN102199592, disclosed method is: the chitosan-arginine negatively charged ion microballoon of take is carrier, add GOD and CAT mixing solutions and glutaraldehyde, through crosslinking reaction, filtration, washing, lyophilize, prepare co-immobilization GOD and CAT microballoon.The advantage of the method is to take full advantage of arginine as " flexible arm ", has realized glucose oxidase and catalatic co-immobilization on " flexible arm ", has reduced the space steric effect of immobilized enzyme.Yet still there are some shortcomings in these methods, such as carrier microballoons surface-area is limited, cannot the too much zymoprotein of load, make enzymatic activity recovery lower.In addition, due to the impact of resistance to mass transfer, the H generating in reaction process
2o
2can not decompose rapidly, thereby cause GOD and CAT inactivation, immobilized enzyme just loses very soon most enzymic activity after using continuously.It is fixation support that the present invention adopts polymethacrylic acid macroporous resin, cost is low, aperture, particle diameter and epoxy group(ing) density are all controlled, research shows, the parameters such as the aperture of resin, particle diameter are very large on the charge capacity impact of enzyme, so polymethacrylic acid macroporous resin can be applicable to greatest extent according to real needs the immobilization of GOD/CAT.The present invention adopts zymoprotein is assembled and is cross-linked, and makes the limited more zymoprotein of carrier surface area energy load.Because carrier has suitable aperture, can overcome the impact of resistance to mass transfer in addition, immobilized enzyme diffusion mass transfer resistance is very little, and speed of reaction is close to using resolvase.
Summary of the invention
Its main purpose of the present invention is to provide and a kind ofly overcomes that immobilization cost was high in the past, complex process, the shortcomings such as the enzyme rate of recovery alive is low provide a kind of mixed immobilized glucose oxidase/catalase (GOD/CAT) microballoon, it is higher that microballoon of the present invention has enzymatic activity recovery, the advantages such as immobilized enzyme is stable, reusable.The gluconic acid that a kind of production process is simple, pollutent is few, product purity is high, energy consumption is low (salt) production method is provided in addition, and immobilized enzyme method is manufactured gluconic acid (salt).Use immobilized enzyme catalysis prepared by the present invention to produce gluconic acid (salt), immobilized enzyme internal divergence resistance to mass transfer is very little, and speed of reaction is close to being used resolvase, and transformation efficiency can reach 100%, and no coupling product generation, sees accompanying drawing 1.
In order to realize object of the present invention, intend adopting following technical scheme:
One aspect of the present invention relates to a kind of mixed immobilized glucose oxidase/catalase microspheres, and it is by prepared the forming of method comprising the steps:
(1) take that to have macroporous structure PGMA microballoon be carrier, the mean pore size of described microballoon is between 100-200nm, take 1-5%(v/v) ethylenediamine solution is amination reagent, by carrier quality: amination reagent volume 1:40 is scattered in carrier in amination reagent, at pH7-9, oscillatory reaction 1-6h at 40-80 ℃, collects amination carrier;
(2) according to glucose oxidase: catalatic vigor, than being the ratio of 1:1-6, is dissolved in GOD and CAT in the phosphoric acid buffer of pH5-8; In enzyme lysate, add carbodiimide and N-hydroxy-succinamide, react 15-60min at 0-8 ℃, then amination carrier is scattered in enzyme liquid to stirring reaction 1-10h at 25-35 ℃;
(3) in step (2) reaction solution, add acetone and the 0.1-2.0%(v/v of 1-5 times of reaction solution volume) linking agent, continue reaction 0.5-1.5h, above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, with phosphoric acid buffer or water washing 3-5 time, the dry mixed immobilized glucose oxidase/catalase microspheres that obtains.
In a preferred embodiment of the present invention, described linking agent is selected from the twain-aldehyde compound linking agents such as glutaraldehyde, Vanillin.
In a preferred embodiment of the present invention, described macroporous structure PGMA microballoon is to obtain by the following method, described method comprises the steps: that the resin for immobilized enzyme is macropore GMA resin, by suspension polymerization, can obtain, the size of controlling microballoon aperture by controlling the consumption of pore-creating agent and linking agent, the size of resin microsphere particle diameter regulates by rotating speed.Preferably, function monomer glycidyl methacrylate (GMA), cross-linking monomer Ethylene glycol dimethacrylate (EDMA), triallyl isocyanide uraturia ester (TAIC), pore-creating agent toluene are mixed, disperse phase is for being suspension stabilizer polyvinyl alcohol (PVA) aqueous solution, then use Diisopropyl azodicarboxylate (AIBN) to carry out polyreaction as initiator, the microballoon generating is through vacuumizing filtration, and with deionized water and ethanol, embathe vacuum-drying.
The present invention also relates to the application of above-mentioned mixed immobilized glucose oxidase/catalase microspheres on the other hand, and gluconic acid or its salt are produced in described being applied as for immobilized enzyme catalysis oxidizing glucose.
In a preferred embodiment of the present invention, described catalytic oxidation process comprises the steps: glucose solution, salt-forming reagent and mixed immobilized glucose oxidase/catalase microspheres add in the reaction vessel with whipping device, temperature of reaction 32-40 ℃, pH 5.5～7.5, ventilation 1:0.14～0.15(v/v/h) (whose volume in whose volume ratio); Oxidization time 12～20 hours, residual sugar termination reaction lower than 0.3% time.
In a preferred embodiment of the present invention, described step is also included in oxidizing reaction and finishes rear recovery mixed immobilized glucose oxidase/catalase microspheres, reuse, preferably, mixed immobilized glucose oxidase/catalase microspheres reuses number of times between 15-20 time.
In a preferred embodiment of the present invention, described salt-forming reagent is selected from calcium carbonate, magnesiumcarbonate, zinc hydroxide.
Mixed immobilized glucose oxidase/catalase microspheres of the present invention can effectively promote the covalently bound of carrier and enzyme molecule, greatly improved immobilization efficiency, and significant prolongation the work-ing life of co-immobilization GOD/CAT, can Reusability time, production cost is low, is conducive to Sustainable development, adopt mixed immobilized glucose oxidase/catalase microspheres of the present invention to prepare gluconate yield high, reaction conditions is gentle, and equipment is simple and easy to get, is a kind of production method of environmental protection.
Accompanying drawing explanation
Fig. 1: enzyme catalysis method is produced calglucon reaction process.
Embodiment
Resin for immobilized enzyme is macropore GMA resin, by suspension polymerization, can obtain, and suspension polymerization is carried out in 250ml there-necked flask, and the size of resin microsphere particle diameter regulates by rotating speed, is rotating speed 500rpm herein.External phase is by function monomer GMA(0.05mol) cross-linking monomer EDMA(0.0125mol), TAIC(0.0125mol), pore-creating agent toluene (0.084 mol) forms.AIBN is as initiator, and its quality is 2% of three kinds of monomer masses.Disperse phase is 1% suspension stabilizer polyvinyl alcohol (PVA).Temperature is controlled at 75 ℃ of 2h, 85 ℃ of 2h.After reaction stops, microballoon is through vacuumizing filtration, and embathes with deionized water and ethanol, vacuum-drying and get final product.The size of controlling microballoon aperture by controlling the consumption of pore-creating agent and linking agent, the size of resin microsphere particle diameter regulates by rotating speed.After reaction stops, microballoon process vacuumizing filtration, and with deionized water and ethanol, embathe vacuum-drying.This resin surface contains abundant epoxide group, and PGMA surface itself has the epoxy group(ing) of high reaction activity, need not just can directly react with amino, hydroxyl, sulfydryl through overactivation.Maximum benefit is to select the suitable carrier of applicable immobilized enzyme by controlling the size of aperture, particle diameter.
Embodiment 1:
A method of preparing mixed immobilized glucose oxidase/catalase microspheres, concrete grammar step is as follows:
Take 2%(v/v) bovine serum albumen solution is amination reagent, carrier mean pore size is 100-200nm, by carrier quality: amination reagent volume 1:40 is scattered in 2%(v/v by carrier) in amination reagent, at pH8.0,200rpm oscillatory reaction 2h at 60 ℃, collects amination carrier.According to the vigor of GOD:CAT, than being the ratio of 1:2, GOD and CAT are dissolved in the phosphoric acid buffer of 0.5mol/l of pH6.Amination carrier is scattered in enzyme liquid, and 30 ℃ are reacted 2h under mixing speed 200rpm, add acetone and the 0.5%(v/v of 2 times of reaction solution volumes in reaction solution) glutaraldehyde, continues reaction 1h.Above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, for the solid substance of collecting, with the 0.01mol/l phosphoric acid buffer of pH7.0, wash 3 times, 4 ℃ dry, just prepares epoxy resin co-immobilization GOD/CAT microballoon, and GOD enzyme activity is 106.8U/g.
Embodiment 2:
Take 2%(v/v) ethylenediamine solution is amination reagent, carrier mean pore size is 100-200nm, by carrier quality: amination reagent volume 1:40 is scattered in 2%(v/v by carrier) in amination reagent, at pH8.0,200rpm oscillatory reaction 2h at 60 ℃, collects amination carrier.According to the vigor of GOD:CAT, than being the ratio of 1:2, GOD and CAT are dissolved in the phosphoric acid buffer of 0.1mol/l of pH6.Amination carrier is scattered in enzyme liquid, and 30 ℃ are reacted 2h under mixing speed 200rpm, add acetone and the 0.5%(v/v of 2 times of reaction solution volumes in reaction solution) glutaraldehyde, continues reaction 1h.Above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, for the solid substance of collecting, with the 0.01mol/l phosphoric acid buffer of pH7.0, wash 3 times, 4 ℃ dry, just prepares epoxy resin co-immobilization GOD/CAT microballoon, and enzyme activity is 111.9U/g.
Embodiment 3:
Take 2%(v/v) ethylenediamine solution is amination reagent, carrier mean pore size is 400-500nm, by carrier quality: amination reagent volume 1:40 is scattered in 2%(v/v by carrier) in amination reagent, at pH8.0,200rpm oscillatory reaction 2h at 60 ℃, collects amination carrier.According to the vigor of GOD:CAT, than being the ratio of 1:2, GOD and CAT are dissolved in the phosphoric acid buffer of 0.1mol/l of pH6.Amination carrier is scattered in enzyme liquid, and 30 ℃ are reacted 2h under mixing speed 200rpm, add acetone and the 0.5%(v/v of 2 times of reaction solution volumes in reaction solution) glutaraldehyde, continues reaction 1h.Above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, for the solid substance of collecting, with the 0.01mol/l phosphoric acid buffer of pH7.0, wash 3 times, 4 ℃ dry, just prepares epoxy resin co-immobilization GOD/CAT microballoon, and enzyme activity is 35.9U/g.
Embodiment 4:
Take 2%(v/v) ethylenediamine solution is amination reagent, carrier mean pore size is 100-200nm, by carrier quality: amination reagent volume 1:40 is scattered in 2%(v/v by carrier) in amination reagent, at pH8.0,200rpm oscillatory reaction 2h at 60 ℃, collects amination carrier.According to the vigor of GOD:CAT, than being the ratio of 1:2, GOD and CAT are dissolved in the phosphoric acid buffer of 0.8mol/l of pH6.In enzyme lysate, add 0.2%(w/v) carbodiimide and 0.2%(w/v) N-hydroxy-succinamide, 4 ℃ reaction 30min.Amination carrier is scattered in enzyme liquid, and 30 ℃ are reacted 2h under mixing speed 200rpm, add acetone and the 1.5%(v/v of 2 times of reaction solution volumes in reaction solution) glutaraldehyde, continues reaction 1h.Above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, for the solid substance of collecting, with the 0.01mol/l phosphoric acid buffer of pH7.0, wash 3 times, 4 ℃ dry, just prepares epoxy resin co-immobilization GOD/CAT microballoon, and enzyme activity is 263U/g.
Embodiment 5:
According to mass percent 20% preparation glucose solution, in the there-necked flask of stirring rake is housed, add glucose solution, calcium carbonate, enzyme amount adds immobilized glucose oxidase and catalase according to 0.6%～1% of glucose amount, be oxidized and carry out in gluconic acid and calcium carbonate and the reaction that produces calglucon simultaneously, oxidizing temperature 32-40 ℃.Ventilation 1:0.14～0.15(v/v/h); Oxidization time 12～20 hours, pH 5.5～7.5, and residual sugar finishes lower than 0.3% time oxidation, after oxidation finishes, reclaims immobilized enzyme, reuses 20 batches of enzyme activities and still has more than 80%.
Immobilized enzyme is compared with resolvase, due to external diffusion in existing, initial reaction rate is slightly low, along with reaction is carried out, owing to using immobilized enzyme reaction to be subject to the impact of product or other condition elements less, speed of reaction, a little more than using resolvase, finally all finishes (as shown in Figure 1) after reaction proceeds to 15h.
The above, be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation of expecting without creative work or replacement, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.
Claims (6)
1. a mixed immobilized glucose oxidase/catalase microspheres, it is by prepared the forming of method comprising the steps:
(1) take that to have macroporous structure PGMA microballoon be carrier, the mean pore size of described microballoon is between 100-200nm, take 1-5%(v/v) ethylenediamine solution is amination reagent, by carrier quality: amination reagent volume 1:40 is scattered in carrier in amination reagent, at pH7-9, oscillatory reaction 1-6h at 40-80 ℃, collects amination carrier;
(2) according to glucose oxidase: catalatic vigor, than being the ratio of 1:1-6, is dissolved in GOD and CAT in the phosphoric acid buffer of pH5-8; In enzyme lysate, add carbodiimide and N-hydroxy-succinamide, react 15-60min at 0-8 ℃, then amination carrier is scattered in enzyme liquid, stirring reaction 1-10h at 25-35 ℃;
(3) in step (2) reaction solution, add acetone and the 0.1-2.0%(v/v of 1-5 times of reaction solution volume) linking agent, continue reaction 0.5-1.5h, above-mentioned reaction solution is filtered, abandon filtrate, collect solid substance, with phosphoric acid buffer or water washing 3-5 time, the dry mixed immobilized glucose oxidase/catalase microspheres that obtains.
2. mixed immobilized glucose oxidase/catalase microspheres according to claim 1, described linking agent is selected from glutaraldehyde, Vanillin.
3. mixed immobilized glucose oxidase/catalase microspheres according to claim 1, described macroporous structure PGMA microballoon is to obtain by the following method, described method comprises the steps: that the resin for immobilized enzyme is macropore GMA resin, by suspension polymerization, can obtain, the size of controlling microballoon aperture by controlling the consumption of pore-creating agent and linking agent, the size of resin microsphere particle diameter regulates by rotating speed.
4. the application of the mixed immobilized glucose oxidase/catalase microspheres described in claim 1-3 any one, gluconic acid or its salt are produced in described being applied as for immobilized enzyme catalysis oxidizing glucose.
5. application according to claim 4, described step is also included in oxidizing reaction and finishes rear recovery mixed immobilized glucose oxidase/catalase microspheres, reuses.
6. application according to claim 5, described step is also included in oxidizing reaction and finishes rear recovery mixed immobilized glucose oxidase/catalase microspheres, reuse, mixed immobilized glucose oxidase/catalase microspheres reuse number of times more than 15 times.
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| CN111419825A (en) * | 2020-06-03 | 2020-07-17 | 烟台大学 | Intelligent response type sustained and controlled release microsphere and preparation method thereof |
| CN114452821A (en) * | 2022-01-26 | 2022-05-10 | 中国科学技术大学 | Bipolar membrane electrodialysis device, method for preparing regenerated alkali by using bipolar membrane electrodialysis device and application of bipolar membrane electrodialysis device |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199592A (en) * | 2011-04-02 | 2011-09-28 | 重庆大学 | Method for preparing mixed immobilized glucose oxidase/catalase microspheres |
-
2012
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199592A (en) * | 2011-04-02 | 2011-09-28 | 重庆大学 | Method for preparing mixed immobilized glucose oxidase/catalase microspheres |
Non-Patent Citations (5)
| Title |
|---|
| 固定化酶法制备葡萄糖酸钙的研究;杜婵娟等;《食品与药品》;20111231;第13卷(第9期);325-327 * |
| 孙志浩主编.交联法.《生物催化工艺学》.2005,308-313. * |
| 杜婵娟等.固定化酶法制备葡萄糖酸钙的研究.《食品与药品》.2011,第13卷(第9期),325-327. * |
| 聚丙烯酸甲酯类大孔树脂对猪胰脂肪酶的固定化研究;谢志东等;《离子交换与吸附》;19951231;第11卷(第1期);24-29 * |
| 谢志东等.聚丙烯酸甲酯类大孔树脂对猪胰脂肪酶的固定化研究.《离子交换与吸附》.1995,第11卷(第1期),24-29. * |
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