CN104651345A - Preparation method of immobilized lipase - Google Patents
Preparation method of immobilized lipase Download PDFInfo
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- CN104651345A CN104651345A CN201310581545.1A CN201310581545A CN104651345A CN 104651345 A CN104651345 A CN 104651345A CN 201310581545 A CN201310581545 A CN 201310581545A CN 104651345 A CN104651345 A CN 104651345A
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- lipase
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- immobilized lipase
- immobilized
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Abstract
The invention discloses a preparation method of immobilized lipase. The preparation method is characterized in that: immobilization Lipase OF is realized via sodium alginate embedding. The immobilized lipase possesses excellent thermal stability; enzyme activity is substantially unchanged after 7h of storage at 60 DEG C; and the immobilized lipase can be used for biodiesel preparation.
Description
Technical field
The present invention relates to a kind of preparation method of immobilized lipase.
Background technology
In lipase water soluble solution, but its natural substrate is as water insoluble in grease.Although adopt suitable organic solvent or emulsifying agent to contribute to the combination of enzyme-to-substrate, still there is technical problem in lipase in the actual utilization of heterogeneous reaction system.Such as, in reactive system remaining enzyme may have a negative impact to the quality of product.The work-ing life of resolvase is relatively short in addition, thus cost is increased.If by lipase immobilization, make it in reaction system, become independent phase, be just fixed in reactor by mechanical means, like this would not pollution products, work-ing life also extends greatly.
In production of biodiesel, the carrier of fixed fat enzyme has the rare phthalein polyimide resin of macropore third, silk, gel, polyurethane foam, biological particles etc., but there is the aperture of glycerine easy plugging particle shape immobilized enzyme, produces the problem of the immobilized enzyme lost of life.For the biofuel of preparation high-quality, low cost, answer lipase immobilization method and the carrier of development of new, wherein the film immobilization of lipase has the incomparable advantage of additive method and receives much concern.
Summary of the invention
The object of this invention is to provide a kind of preparation method of immobilized lipase, adopt cheap, active high free-fat enzyme Lipause OF as catalyzer, solve catalyzer and be subject to alcohol effect and the problem such as inactivation and catalyzer life-time service.
For achieving the above object, technical scheme of the present invention is as follows:
A certain amount of LipaSe OF and sodium alginate (concentration 1%-3%) are dissolved in phosphoric acid buffer (pH value 6.0-8.0) respectively, fully dissolve, merge two solution, fully stir, mix; Dropwise be added dropwise in calcium chloride solution (0.02-0.15mol/L) with microsyringe, under room temperature, fix 20-60min; Filter, at 35 DEG C, vacuum tightness is dry in the vacuum drying oven of 0.09MPa.
The immobilized enzyme that the present invention adopts sodium alginate to embed method obtained, have good thermostability, at 60 DEG C of temperature, keep 7h, its enzyme activity is substantially constant.
Embodiment
Embodiment 1
A certain amount of Lipase OF and sodium alginate (concentration 2%) are dissolved in phosphoric acid buffer (pH value 7.0) respectively, fully dissolve, merge two solution, fully stir, mix; Dropwise be added dropwise in calcium chloride solution (0.05mol/L) with microsyringe, under room temperature, fix 30min; Filter, at 35 DEG C, vacuum tightness is dry in the vacuum drying oven of 0.09MPa.Enzyme is lived and is reached 1810U/g.
Embodiment 2
A certain amount of Lipase OF and sodium alginate (concentration 2%) are dissolved in phosphoric acid buffer (pH value 7.5) respectively, fully dissolve, merge two solution, fully stir, mix; Dropwise be added dropwise in calcium chloride solution (0.05mol/L) with microsyringe, under room temperature, fix 40min; Filter, at 35 DEG C, vacuum tightness is dry in the vacuum drying oven of 0.09MPa.Enzyme is lived and is reached 2300U/g.
Above embodiment is only in order to illustrate technical scheme of the present invention, but not be limited, although with reference to above-described embodiment to invention has been detailed description, for the person of ordinary skill of the art, still can the technical scheme described in above-described embodiment be modified, or equivalent replacement is carried out to wherein portion of techniques feature, and these amendments or replace, do not make the essence of appropriate technical solution depart from the spirit and scope of technical scheme of the presently claimed invention.
Claims (1)
1. the preparation method of an immobilized lipase, it is characterized in that: a certain amount of LipaSe OF and sodium alginate (concentration 1%-3%) are dissolved in phosphoric acid buffer (pH value 6.0-8.0) respectively, fully dissolve, merge two solution, abundant stirring, mixes; Dropwise be added dropwise in calcium chloride solution (0.02-0.15mol/L) with microsyringe, under room temperature, fix 20-60min; Filter, at 35 DEG C, vacuum tightness is dry in the vacuum drying oven of 0.09MPa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310581545.1A CN104651345A (en) | 2013-11-19 | 2013-11-19 | Preparation method of immobilized lipase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310581545.1A CN104651345A (en) | 2013-11-19 | 2013-11-19 | Preparation method of immobilized lipase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104651345A true CN104651345A (en) | 2015-05-27 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN201310581545.1A Pending CN104651345A (en) | 2013-11-19 | 2013-11-19 | Preparation method of immobilized lipase |
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| CN (1) | CN104651345A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105274157A (en) * | 2015-12-01 | 2016-01-27 | 河北工业大学 | Method for producing biodiesel by utilizing immobilized lipase and adopting static emulsion method |
| CN108624579A (en) * | 2018-04-20 | 2018-10-09 | 中国科学院南海海洋研究所 | A method of eliminating immobilized lipase excess epoxy group |
| WO2022151024A1 (en) * | 2021-01-13 | 2022-07-21 | 安徽金禾实业股份有限公司 | Immobilization method of liquid lipase and preparation method of sucrose-6-acetate |
-
2013
- 2013-11-19 CN CN201310581545.1A patent/CN104651345A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105274157A (en) * | 2015-12-01 | 2016-01-27 | 河北工业大学 | Method for producing biodiesel by utilizing immobilized lipase and adopting static emulsion method |
| CN105274157B (en) * | 2015-12-01 | 2018-07-20 | 河北工业大学 | A kind of method of static state emulsion method immobilized lipase production biodiesel |
| CN108624579A (en) * | 2018-04-20 | 2018-10-09 | 中国科学院南海海洋研究所 | A method of eliminating immobilized lipase excess epoxy group |
| WO2022151024A1 (en) * | 2021-01-13 | 2022-07-21 | 安徽金禾实业股份有限公司 | Immobilization method of liquid lipase and preparation method of sucrose-6-acetate |
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Legal Events
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| PB01 | Publication | ||
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150527 |