CN1885024A - Method for fixing enzyme in glucose sensor - Google Patents
Method for fixing enzyme in glucose sensor Download PDFInfo
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- CN1885024A CN1885024A CN 200610028189 CN200610028189A CN1885024A CN 1885024 A CN1885024 A CN 1885024A CN 200610028189 CN200610028189 CN 200610028189 CN 200610028189 A CN200610028189 A CN 200610028189A CN 1885024 A CN1885024 A CN 1885024A
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- konjaku
- enzyme
- glucose sensor
- deacetylate
- sol
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for fixing the enzyme of glucose sensor, which is characterized in that: first preparing sol-gel without acetyl konjak glucans; then using said sol-gel to fix the glucose oxidase on the surface of electrode to prepare the glucose sensor. The inventive film has better waterproof property, strong adhesive, and better biological compatibility; then the desacetyl konjak glucans is used to fix the glucose oxydase at the surface of electrode, to prepare relative sensor. Said sensor has high stability and long service life. The invention can improve the inactivation and leakage of enzyme, to improve stability and service life.
Description
Technical field
The present invention relates to the fixing means of enzyme in a kind of biology sensor, more particularly relate to the fixing means of enzyme in a kind of glucose sensor.
Background technology
Enzyme fixedly is a committed step in the biology sensor preparation process.Normally used enzyme immobilization method has that absorption method, covalent bond are legal, cross-linking method, polymkeric substance investment, and wherein the polymkeric substance investment is the most general.Natural polymer has good bioaffinity, can make enzyme keep higher activity.But the natural polymer ubiquity of current use and the cohesiveness of electrode are not high, contact the shortcoming of easy water absorption and swelling with water, easily cause coming off and the leakage of enzyme of enzyme membrane.Konjaku glucomannan obtains the deacetylate konjaku glucomannan after the deacetylate modification is handled, its sol-gel has strong cohesive force, and the film that is become can water absorption and swelling, has good water resisting property.But at present the deacetylate konjaku glucomannan is applied to that the immobilization of enzyme still is not reported in the field of biosensors.
Summary of the invention
Technical matters to be solved by this invention is that the konjaku glucomannan through the deacetylate modification is applied to the fixing of enzyme in the biology sensor, and the fixing means of enzyme in a kind of new glucose sensor is provided.
The technical solution used in the present invention: the fixing means of enzyme in a kind of glucose sensor; it is characterized in that: the first sol-gel of preparation deacetylate konjaku glucomannan is fixed on electrode surface with the sol-gel of described deacetylate konjaku glucomannan with glucose oxidase then and prepares glucose sensor.
The fixing means of enzyme in described a kind of glucose sensor is characterized in that:
A. it is 0.1%～0.3% Na that the konjaku powder 5～15g that purifying is crossed joins 1L concentration
2CO
3In the solution, agitating solution dissolves fully until konjaku powder, room temperature ageing 24～48 hours, and it is standby to obtain deacetylate konjaku glucomannan sol-gel;
The b.Pt electrode is polished through abrasive paper for metallograph; 0.05 the alumina powder of μ m polishing; ultrasonic cleaning; airing; compound concentration is the glucose oxidase solution of 900～1100U/ml, and 50～100 μ l deacetylate konjaku glucomannan sol-gels that pipette 5～10 μ l glucose oxidase solutions and step a preparation with micropipettor mix, and pipettes mixed liquor 2～5 μ l at last and drips and be applied on the Pt electrode; drying at room temperature 24～48 hours promptly makes described glucose sensor.
The purifying of the described konjaku powder of step a is that konjaku powder is added in the beaker, adds 40% ethanol then, stirs 10min, suction filtration, filter residue re-treatment three times; Repeat to stir 10min, suction filtration, the step that the filter residue re-treatment is three times with 60%, 95% and 100% ethanol respectively again; The gained filter residue changes in the surface plate, and normal temperature is placed and promptly got the konjaku powder that purifying is crossed in 48 hours.
Beneficial effect of the present invention, acetyl group on the konjaku glucomannan side chain short-chain branch and the hydroxyl on the saccharide residue form ester bond, make konjaku glucomannan form the double-spiral structure with space, stop the konjaku glucomannan long-chain close mutually.Therefore, the easy water absorption and swelling of konjaku glucomannan.Under alkali condition, ester bond generation hydrolysis, promptly deacetylate generates the deacetylate konjaku glucomannan.Because deacetylate konjaku glucomannan sugar chain is naked shape, its long-chain becomes can be close mutually.So the intermolecular formation hydrogen bond of part and produce crystallization is tubercle point with the crystallization and then formed the reticulate texture body.Glucose oxidase is embedded in the reticulate texture body.The enzyme membrane of this method preparation can water absorption and swelling, has good water resisting property.The sensor of the present invention preparation is the glucose solution replicate determination 50 times of 5mmol/L to concentration, and RSD is 2.4%, has advantages of higher stability.In sensor three months after making (the time spent is not stored in the refrigerator), be the glucose solution replication 12 times (jede Woche is measured once) of 5mmol/L to concentration, its response current does not almost descend, and shows that this sensor has good serviceable life.Sensor good stable and long serviceable life illustrate that the present invention has improved the leakage of coming off of enzyme membrane and enzyme effectively.
Embodiment
Below by embodiment the present invention is described in further detail; the fixing means of enzyme in a kind of glucose sensor; the first sol-gel of preparation deacetylate konjaku glucomannan is fixed on electrode surface with the sol-gel of described deacetylate konjaku glucomannan with glucose oxidase then and prepares glucose sensor.More specifically, the fixing means of enzyme is in described a kind of glucose sensor: it is 0.1%～0.3% Na that konjaku powder 5～15g that a. crosses purifying joins 1L concentration
2CO
3In the solution, agitating solution dissolves fully until konjaku powder, room temperature ageing 24～48 hours, and it is standby to obtain deacetylate konjaku glucomannan sol-gel; The b.Pt electrode is polished through abrasive paper for metallograph; 0.05 the alumina powder of μ m polishing; ultrasonic cleaning; airing; compound concentration is the glucose oxidase solution of 900～1100U/ml, and 50～100 μ l deacetylate konjaku glucomannan sol-gels that pipette 5～10 μ l glucose oxidase solutions and step a preparation with micropipettor mix, and pipettes mixed liquor 2～5 μ l at last and drips and be applied on the Pt electrode; drying at room temperature 24～48 hours promptly makes described glucose sensor.The purifying of the described konjaku powder of step a is that konjaku powder is added in the beaker, adds 40% ethanol then, stirs 10min, suction filtration, filter residue re-treatment three times; Repeat to stir 10min, suction filtration, the step that the filter residue re-treatment is three times with 60%, 95% and 100% ethanol respectively again; The gained filter residue changes in the surface plate, and normal temperature is placed and promptly got the konjaku powder that purifying is crossed in 48 hours.
Embodiment 1:
The first step: the purifying of konjaku powder
Konjaku powder is added in the beaker, add 40% ethanol then, stir 10min, suction filtration, filter residue re-treatment three times.Repeat above-mentioned steps with 60%, 95% and 100% ethanol respectively again.The gained filter residue changes in the surface plate, and normal temperature is placed and promptly got the good konjaku powder of purifying in 48 hours.
Second step: the preparation of deacetylate konjaku glucomannan sol-gel
Taking by weighing konjaku powder 8g that purifying crosses, to join 1L concentration be 0.2% Na
2CO
3In the solution, agitating solution dissolves fully until konjaku powder.Room temperature ageing 24 hours promptly gets deacetylate konjaku glucomannan sol-gel.
The 3rd step: enzyme is fixing in the glucose sensor
The Pt electrode is polished through abrasive paper for metallograph, the alumina powder polishing of 0.05 μ m, ultrasonic cleaning, airing.Compound concentration is glucose oxidase (GOD) solution of 1000U/ml.Pipetting 5 these solution of μ l with micropipettor mixes with 50 μ l deacetylate konjak portuguese gansu polyose sugar sols.Pipette above-mentioned mixed liquor 2 μ l again and drip and be applied on the Pt electrode, drying at room temperature 24 hours promptly makes sensor.
Above said content only is the basic explanation of the present invention under conceiving, and according to any equivalent transformation that technical scheme of the present invention is done, all should belong to protection scope of the present invention.
Claims (3)
1. the fixing means of enzyme in the glucose sensor; it is characterized in that: the first sol-gel of preparation deacetylate konjaku glucomannan is fixed on electrode surface with the sol-gel of described deacetylate konjaku glucomannan with glucose oxidase then and prepares glucose sensor.
2. according to the fixing means of enzyme in the described a kind of glucose sensor of claim 1, it is characterized in that:
A. it is 0.1%～0.3% Na that the konjaku powder 5～15g that purifying is crossed joins 1L concentration
2CO
3In the solution, agitating solution dissolves fully until konjaku powder, room temperature ageing 24～48 hours, and it is standby to obtain deacetylate konjaku glucomannan sol-gel;
The b.Pt electrode is polished through abrasive paper for metallograph; 0.05 the alumina powder of μ m polishing; ultrasonic cleaning; airing; compound concentration is the glucose oxidase solution of 900～1100U/ml, and 50～100 μ l deacetylate konjaku glucomannan sol-gels that pipette 5～10 μ l glucose oxidase solutions and step a preparation with micropipettor mix, and pipettes mixed liquor 2～5 μ l at last and drips and be applied on the Pt electrode; drying at room temperature 24～48 hours promptly makes described glucose sensor.
3. according to the fixing means of enzyme in the described a kind of glucose sensor of claim 2, it is characterized in that: the purifying of the described konjaku powder of step a is that konjaku powder is added in the beaker, adds 40% ethanol then, stirs 10min, suction filtration, filter residue re-treatment three times; Repeat to stir 10min, suction filtration, the step that the filter residue re-treatment is three times with 60%, 95% and 100% ethanol respectively again; The gained filter residue changes in the surface plate, and normal temperature is placed and promptly got the konjaku powder that purifying is crossed in 48 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610028189 CN1885024A (en) | 2006-06-27 | 2006-06-27 | Method for fixing enzyme in glucose sensor |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610028189 CN1885024A (en) | 2006-06-27 | 2006-06-27 | Method for fixing enzyme in glucose sensor |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100582228C (en) * | 2007-03-29 | 2010-01-20 | 中国农业科学院农产品加工研究所 | Immobilization lipase, and preparation method |
| CN109030601A (en) * | 2018-08-10 | 2018-12-18 | 上海应用技术大学 | A kind of electric potential type organophosphorus pesticide biosensor enzyme membrane and preparation method thereof |
| CN115780215A (en) * | 2022-06-06 | 2023-03-14 | 东莞市通科电子有限公司 | Method for protecting tin coating of pin of electronic component |
-
2006
- 2006-06-27 CN CN 200610028189 patent/CN1885024A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100582228C (en) * | 2007-03-29 | 2010-01-20 | 中国农业科学院农产品加工研究所 | Immobilization lipase, and preparation method |
| CN109030601A (en) * | 2018-08-10 | 2018-12-18 | 上海应用技术大学 | A kind of electric potential type organophosphorus pesticide biosensor enzyme membrane and preparation method thereof |
| CN115780215A (en) * | 2022-06-06 | 2023-03-14 | 东莞市通科电子有限公司 | Method for protecting tin coating of pin of electronic component |
| CN115780215B (en) * | 2022-06-06 | 2023-12-19 | 东莞市通科电子有限公司 | Method for protecting tin plating layer of pin of electronic component |
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