CN106754856A - A kind of method that co-immobilization glucose oxidase and catalase prepare ferrous gluconate - Google Patents
A kind of method that co-immobilization glucose oxidase and catalase prepare ferrous gluconate Download PDFInfo
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- CN106754856A CN106754856A CN201611083020.5A CN201611083020A CN106754856A CN 106754856 A CN106754856 A CN 106754856A CN 201611083020 A CN201611083020 A CN 201611083020A CN 106754856 A CN106754856 A CN 106754856A
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- catalase
- glucose oxidase
- immobilization
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- ferrous
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/58—Aldonic, ketoaldonic or saccharic acids
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- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
- C12Y101/03004—Glucose oxidase (1.1.3.4)
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- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01006—Catalase (1.11.1.6)
Abstract
The invention discloses a kind of method that co-immobilization glucose oxidase and catalase prepare ferrous gluconate.The present invention is with glucose as substrate, ferrous gluconate is prepared using glucolactone, shitosan and calcium alginate co-immobilization glucose oxidase and catalase catalytic oxidation of glucose technology, the technology significantly improves enzyme activity and reduces step of converting, sugared conversion ratio is set to be largely increased, energy consumption has been saved, production cost has been reduced.
Description
Technical field
The present invention relates to a kind of preparation method of ferrous gluconate, more particularly to a kind of co-immobilization glucose oxidase
With the method that catalase prepares ferrous gluconate.
Background technology
Ferrous gluconate is a kind of good iron supplementary, can be used with medicine or in the form of food additives, to scarce
Iron person brings healthy good news.Human body iron deficiency can make decreased hemoglobin, and alimentary anemia occurs.Human body is to the absorption of iron by many
Factor influences, wherein, it is soluble low molecule ferrous iron that iron must be digested（It is ferrous）Compound can just accelerate absorb, and
Organic acid（Such as gluconic acid）Play a part of to promote body to absorb iron.
Ferrous gluconate is that China copys a kind of external new drug developed at the end of the eighties.China's food service industry also uses it
To make new food additives.FAO (Food and Agriculture Organization of the United Nation)/the World Health Organization (F A O/WH O) food additives expert
Committee member will be considered that ferrous gluconate is a kind of iron supplementary with " serviceability, security, necessity ".With clinically institute at present
Ferrous sulfate compares, and it has, and stability is high, in good taste, toxicity is low, Small side effects, being easily absorbed by the body utilizes etc. significantly
Advantage.
The production method of current ferrous gluconate mainly has the side such as legal, oxidation exchange process, acidolysis-exchange process in oxidation
Method, four kinds of methods cut both ways, and with biological and enzyme the reach of science, Production by Enzymes is subject to vast manufacturer and consumer
Concern.
The content of the invention
The object of the invention is exactly to make up the defect of prior art, there is provided a kind of co-immobilization glucose oxidase and mistake
The method that hydrogen oxide enzyme prepares ferrous gluconate.The present invention is using glucolactone, calcium alginate and shitosan to grape
Carbohydrate oxidase and being fixed of catalase, prepare immobilised enzymes, glucose are aoxidized by immobilised enzymes, in
With obtain ferrous gluconate.The invented technology is simple, is capable of achieving the continuous prodution of ferrous gluconate, and greatly facilitate
The techniques such as separation, purifying, the drying of follow-up ferrous gluconate.
The present invention is achieved by the following technical solutions：
（1）The co-immobilization of glucose oxidase and catalase：Take glucose oxidase/catalase（U/U）=1:1,
Under conditions of pH7.0, glucolactone, calcium alginate and shitosan is added to be crosslinked and esterification, course of reaction
Middle stirring, reaction is prepared into the microballoon of a diameter of 0.1-0.12cm after terminating；
（2）Microballoon is fixed in enzymatic vessel, with glucose as substrate, hydroxide is used in being fixed enzyme reaction in course of reaction
Sodium adjust pH=6.5, it is neutralized after obtain sodium gluconate；
（3）After pH is 6.5 stabilizations, sodium gluconate solution obtains ferrous gluconate by ferrous ion exchange column, then through from
Son is exchanged, crystallized, being dried to obtain ferrous gluconate.
The method that a kind of described co-immobilization glucose oxidase and catalase prepare ferrous gluconate, it is special
Levying is, the step（1）In, the condition of crosslinking and esterification is：Reaction time common 8h, wherein 0-4h, 35-40 DEG C of temperature,
4-8h, 4-8 DEG C of temperature.
The method that a kind of described co-immobilization glucose oxidase and catalase prepare ferrous gluconate, it is special
Levying is, the step（1）In, the mass fraction of glucolactone, calcium alginate and shitosan is respectively：Glucolactone
1 part, calcium alginate 8-15 parts, shitosan 15-20 parts.
The method that a kind of described co-immobilization glucose oxidase and catalase prepare ferrous gluconate, it is special
Levying is, the step（2）In, the mass concentration of glucose is 10-25%.
The method that a kind of described co-immobilization glucose oxidase and catalase prepare ferrous gluconate, it is special
Levying is, the step（2）In, the condition of immobilised enzymes reaction is 40 DEG C of temperature, and rotating speed 150rpm, dissolved oxygen reaches 70%-90%.
The present invention has advantages below：
（1）Compared with traditional ferrous gluconate preparation method, with energy-conservation, the advantage of production cost is reduced, and substantially not
Three-waste pollution is produced, meets green production process requirement；
（2）Ferrous gluconate purity prepared by the technique is high, reaches 99.9%, and content of beary metal is not detected, and product quality is higher than
USP standard.
Specific embodiment
The present embodiment is further illustrating rather than the limitation to this patent to this patent.
Embodiment 1
（1）The co-immobilization of glucose oxidase and catalase：Take glucose oxidase/catalase（U/U）=1:1,
Under conditions of pH7.0, glucolactone, calcium alginate and shitosan is added to be crosslinked and esterification, course of reaction
Middle stirring, reaction is prepared into the microballoon of a diameter of 0.1-0.12cm after terminating；
（2）Microballoon is fixed in enzymatic vessel, with glucose as substrate, hydroxide is used in being fixed enzyme reaction in course of reaction
Sodium adjust pH=6.5, it is neutralized after obtain sodium gluconate；
（3）After pH is 6.5 stabilizations, sodium gluconate solution obtains ferrous gluconate by ferrous ion exchange column, then through from
Son is exchanged, crystallized, being dried to obtain ferrous gluconate.
The step（1）In, the condition of crosslinking and esterification is：Reaction time common 8h, wherein 0-4h, 35 DEG C of temperature,
4-8h, 4 DEG C of temperature；The mass fraction of glucolactone, calcium alginate and shitosan is respectively：1 part of glucolactone, sea
8 parts of calcium alginate, 15 parts of shitosan.
The step（2）In, the mass concentration of glucose is 10%；The condition of immobilised enzymes reaction is 40 DEG C of temperature, rotating speed
150rpm, dissolved oxygen reaches 70%%.
Embodiment 2
（1）The co-immobilization of glucose oxidase and catalase：Take glucose oxidase/catalase（U/U）=1:1,
Under conditions of pH7.0, glucolactone, calcium alginate and shitosan is added to be crosslinked and esterification, course of reaction
Middle stirring, reaction is prepared into the microballoon of a diameter of 0.1-0.12cm after terminating；
（2）Microballoon is fixed in enzymatic vessel, with glucose as substrate, hydroxide is used in being fixed enzyme reaction in course of reaction
Sodium adjust pH=6.5, it is neutralized after obtain sodium gluconate；
（3）After pH is 6.5 stabilizations, sodium gluconate solution obtains ferrous gluconate by ferrous ion exchange column, then through from
Son is exchanged, crystallized, being dried to obtain ferrous gluconate.
The step（1）In, the condition of crosslinking and esterification is：Reaction time common 8h, wherein 0-4h, 38 DEG C of temperature,
4-8h, 6 DEG C of temperature；The mass fraction of glucolactone, calcium alginate and shitosan is respectively：1 part of glucolactone, sea
12 parts of calcium alginate, 18 parts of shitosan.
The step（2）In, the mass concentration of glucose is 15%；The condition of immobilised enzymes reaction is 40 DEG C of temperature, rotating speed
150rpm, dissolved oxygen reaches 80%.
Embodiment 3
（1）The co-immobilization of glucose oxidase and catalase：Take glucose oxidase/catalase（U/U）=1:1,
Under conditions of pH7.0, glucolactone, calcium alginate and shitosan is added to be crosslinked and esterification, course of reaction
Middle stirring, reaction is prepared into the microballoon of a diameter of 0.1-0.12cm after terminating；
（2）Microballoon is fixed in enzymatic vessel, with glucose as substrate, hydroxide is used in being fixed enzyme reaction in course of reaction
Sodium adjust pH=6.5, it is neutralized after obtain sodium gluconate；
（3）After pH is 6.5 stabilizations, sodium gluconate solution obtains ferrous gluconate by ferrous ion exchange column, then through from
Son is exchanged, crystallized, being dried to obtain ferrous gluconate.
The step（1）In, the condition of crosslinking and esterification is：Reaction time common 8h, wherein 0-4h, 40 DEG C of temperature,
4-8h, 8 DEG C of temperature；The mass fraction of glucolactone, calcium alginate and shitosan is respectively：1 part of glucolactone, sea
15 parts of calcium alginate, 20 parts of shitosan.
The step（2）In, the mass concentration of glucose is 25%；The condition of immobilised enzymes reaction is 40 DEG C of temperature, rotating speed
150rpm, dissolved oxygen reaches 90%.
Claims (5)
1. a kind of method that co-immobilization glucose oxidase and catalase prepare ferrous gluconate, it is characterised in that press
Following step is obtained：
（1）The co-immobilization of glucose oxidase and catalase：Take glucose oxidase/catalase（U/U）=1:1,
Under conditions of pH7.0, glucolactone, calcium alginate and shitosan is added to be crosslinked and esterification, course of reaction
Middle stirring, reaction is prepared into the microballoon of a diameter of 0.1-0.12cm after terminating；
（2）Microballoon is fixed in enzymatic vessel, with glucose as substrate, hydroxide is used in being fixed enzyme reaction in course of reaction
Sodium adjust pH=6.5, it is neutralized after obtain sodium gluconate；
（3）After pH is 6.5 stabilizations, sodium gluconate solution obtains ferrous gluconate by ferrous ion exchange column, then through from
Son is exchanged, crystallized, being dried to obtain ferrous gluconate.
2. a kind of co-immobilization glucose oxidase and catalase according to claim 1 prepare gluconic acid
Ferrous method, it is characterized in that, the step（1）In, the condition of crosslinking and esterification is：Reaction time common 8h, wherein 0-
4h, 35-40 DEG C of temperature, 4-8h, 4-8 DEG C of temperature.
3. a kind of co-immobilization glucose oxidase and catalase according to claim 1 prepare gluconic acid
Ferrous method, it is characterized in that, the step（1）In, the mass fraction difference of glucolactone, calcium alginate and shitosan
For：1 part, calcium alginate 8-15 parts, shitosan 15-20 parts of glucolactone.
4. a kind of co-immobilization glucose oxidase and catalase according to claim 1 prepare gluconic acid
Ferrous method, it is characterized in that, the step（2）In, the mass concentration of glucose is 10-25%.
5. a kind of co-immobilization glucose oxidase and catalase according to claim 1 prepare gluconic acid
Ferrous method, it is characterized in that, the step（2）In, the condition of immobilised enzymes reaction is 40 DEG C of temperature, and rotating speed 150rpm is molten
Oxygen reaches 70%-90%.
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| CN201611083020.5A CN106754856A (en) | 2016-11-30 | 2016-11-30 | A kind of method that co-immobilization glucose oxidase and catalase prepare ferrous gluconate |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102943069A (en) * | 2012-11-27 | 2013-02-27 | 北京化工大学 | Co-immobilization glucose oxidase/catalase microspheres and application thereof in production of gluconic acid or gluconic salt |
| CN103602710A (en) * | 2013-09-30 | 2014-02-26 | 山东西王糖业有限公司 | Method of preparing calcium gluconate by composite immobilized enzyme |
| CN104263715A (en) * | 2014-09-26 | 2015-01-07 | 东北农业大学 | Method for immobilizing phospholipase A2 by calcium alginate-chitosan |
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2016
- 2016-11-30 CN CN201611083020.5A patent/CN106754856A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102943069A (en) * | 2012-11-27 | 2013-02-27 | 北京化工大学 | Co-immobilization glucose oxidase/catalase microspheres and application thereof in production of gluconic acid or gluconic salt |
| CN103602710A (en) * | 2013-09-30 | 2014-02-26 | 山东西王糖业有限公司 | Method of preparing calcium gluconate by composite immobilized enzyme |
| CN104263715A (en) * | 2014-09-26 | 2015-01-07 | 东北农业大学 | Method for immobilizing phospholipase A2 by calcium alginate-chitosan |
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