CN104894094A - Enzyme immobilization method and enzyme preparation - Google Patents
Enzyme immobilization method and enzyme preparation Download PDFInfo
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- CN104894094A CN104894094A CN201510250417.8A CN201510250417A CN104894094A CN 104894094 A CN104894094 A CN 104894094A CN 201510250417 A CN201510250417 A CN 201510250417A CN 104894094 A CN104894094 A CN 104894094A
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- enzyme
- immobilization
- lipase
- aspergillus niger
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Abstract
The invention discloses an enzyme immobilization method and an enzyme preparation, wherein the enzyme immobilization method comprises the following steps: (1) dispersing ferroferric oxide nanoparticles in a Tris-HCL buffer solution, and preparing a substrate solution; (2) adding dopamine hydrochloride to the substrate solution obtained in the step (1), adjusting pH to 8.0-9.5, carrying out vibration for 1-90 min at room temperature, then repeatedly swashing by deionized water, and drying; (3) carrying out vibration immobilization of the material obtained in the step (2) with an enzyme in a phosphate buffer solution with the immobilization temperature of 0-4 DEG C and the immobilization time of 1-12 h; and (4) freezing and drying the immobilized enzyme obtained in the step (3), and thus obtaining the product. The enzyme immobilization method has the characteristics of high immobilization efficiency of immobilization of aspergillus niger lipase, high enzyme activity recovery rate, simple and convenient operation, mild conditions and the like.
Description
Technical field
The present invention relates to a kind of lipase from Aspergillus Niger process for fixation.Use nano ferriferrous oxide material to be base material, by biomimetic material Dopamine HCL autohemagglutination synthesis poly-Dopamine HCL, enzyme is bonded in substrate surface, thus the zymin of being fixed.
Background technology
In traditional sense, immobilization refers to that enzyme is spatially limited in around some water-insoluble carriers by the method for Bian physics or chemistry, limits its freely movable activity still not affecting biological catalyst.Traditional immobilization technology is mainly divided into four kinds: absorption method, entrapping method, covalent coupling method, crosslinking.The immobilization efficiency (being less than 40%) of usual absorption method and entrapping method and the enzyme rate of recovery alive lower (being less than 20%), crosslinking is owing to adding linking agent, and the loss that enzyme is lived is larger.Along with the progress of biotechnology and material science and technology, immobilization technology is faced with a very important transition.Increasing type material is used for enzyme immobilizatio, and especially nano material has more and more important application with the physicochemical property of its uniqueness in immobilization field.Traditional porous material cannot improve its charge capacity further, is because inevitably reduce the diameter of micropore while of increasing carrier specific surface area, thus reduces its carrying capacity.And nano material has very large specific surface area, therefore there is good carrying capacity.But the surface of these carriers often lacks the combination that active functional group is unfavorable for albumen.
Summary of the invention
The object of the invention is to the defect overcoming prior art, provide a kind of simply based on the process for fixation of bionical bonding, define the immobilized enzyme of nano ferriferrous oxide-poly-Dopamine HCL-lipase from Aspergillus Niger amine three-decker.Have that Immobilization of lipase from Aspergillus Niger immobilization efficiency is high, the enzyme rate of recovery of living is high, easy and simple to handle, the features such as mild condition.
Object of the present invention is achieved through the following technical solutions:
A kind of enzyme immobilization method, comprises the steps:
(1) ferriferrous oxide nano-particle is dispersed in the buffered soln of Tris-HCL, is configured to substrate solution;
(2) add dopamine hydrochloride by the substrate solution of gained in (1), adjust pH to be 8.0 to 9.5, at room temperature shake 1 ~ 90min, then repeatedly use deionized water rinsing, dry;
(3) material (2) obtained carries out concussion immobilization with enzyme in phosphoric acid buffer, and immobilization temperature is 0 ~ 4 DEG C, immobilization time 1 ~ 12h;
(4) by after the immobilized enzyme lyophilize of gained in (3).
Described in step (2), pH is 8.5, and in step (3), the pH of phosphoric acid buffer is 8.0.
Iron dichloride tetrahydrate and ferric chloride hexahydrate are obtained by ammoniacal liquor co-precipitation by the ferriferrous oxide nano-particle in described step (1).
The mass ratio of described ferriferrous oxide nano-particle and Dopamine HCL is 1:(0.5 ~ 5).
In described step (2), (3), concussion speed is 100 ~ 500rpm.
Enzyme in described step (2) is lipase or proteolytic enzyme.
Described lipase is lipase from Aspergillus Niger, the fermented liquid that lipase from Aspergillus Niger is lipase from Aspergillus Niger enzyme powder, born of the same parents contain lipase from Aspergillus Niger after producing lipase from Aspergillus Niger or cell wall breaking outward.
Zymin prepared by aforesaid method, its optimal temperature is 20 ~ 60 DEG C, and pH is 6.5 ~ 9.0.
Preferably, the optimal temperature of this zymin is 35 ~ 50 DEG C, and pH is 7.0 ~ 8.0.
The principle of the invention: can ferriferrous oxide nano-particle be obtained by Iron dichloride tetrahydrate and ferric chloride hexahydrate and ammoniacal liquor co-precipitation, Dopamine HCL is under weakly alkaline (pH=8.5) condition being similar to seawater simultaneously, auto-polymerization can form poly-Dopamine HCL, hydroxyl in catechol group in poly-dopamine molecule can form coordinate bond with iron atom, its reactive force is very large, poly-dopamine molecule is made can be tightly wrapped in nano ferriferrous oxide surface, catechol group simultaneously in poly-dopamine molecule is as easy as rolling off a log is in the basic conditions oxidized to adjacent benzene diquinone, thus can react with the amino in enzyme molecule or sulfydryl, by protein firm " bonding " at ferriferrous oxide nano-particle matrix surface, achieve enzyme immobilizatio.
The present invention has the following advantages relative to prior art:
(1) invention use simple bionical adhering technique, in immobilized enzyme process, do not use the bifunctional reagent such as glutaraldehyde, formaldehyde, cost is lower, reaction temperature and, can keep largely enzyme live (60% ~ 90%), improve enzyme live the rate of recovery.
(2) the present invention effectively improves the adhesive capacity of natural carrier to albumen, enzyme immobilizatio better effects if by the modification of poly-Dopamine HCL.
(3) immobilized enzyme method of the present invention, can be used in a series of enzyme containing enriching amino or mercapto groups such as lipase, proteolytic enzyme, and has the potential quality as pharmaceutical grade protein carrier.
Accompanying drawing explanation
Zymin relative enzyme differing temps under the alive curve of Fig. 1 for obtaining in embodiment 1.
Zymin relative enzyme under different pH the alive curve of Fig. 2 for obtaining in embodiment 1.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 37.5mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 12h under the condition of ice bath, immobilized enzyme efficiency is 74.1%, the enzyme rate of recovery alive is 56.3%, and the charge capacity of this carrier is 134mg enzyme/g carrier.
Embodiment 2
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 30mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 12h under the condition of ice bath, immobilized enzyme efficiency is 63.2%, the enzyme rate of recovery alive is 48.8%, and the charge capacity of this carrier is 114mg enzyme/g carrier.
Embodiment 3
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 12h under the condition of ice bath, immobilized enzyme efficiency is 64.1%, the enzyme rate of recovery alive is 46.2%, and the charge capacity of this carrier is 115mg enzyme/g carrier.
Embodiment 4
40mg nano ferriferrous oxide is dispersed in the Tris-HCL buffered soln of 15mL pH 8.5, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 30min under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 12h under the condition of ice bath, immobilized enzyme efficiency is 68.8%, the enzyme rate of recovery alive is 52.7%, and the charge capacity of this carrier is 124mg enzyme/g carrier.
Embodiment 5
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1.5h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 12h under the condition of ice bath, immobilized enzyme efficiency is 69.7%, the enzyme rate of recovery alive is 54.5%, and the charge capacity of this carrier is 126mg enzyme/g carrier.
Embodiment 6
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1.5h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, and then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 9h under the condition of ice bath, immobilized enzyme efficiency is 58.0%, and the enzyme rate of recovery alive is 30.3%, and the charge capacity of this carrier is 105mg enzyme/g carrier.
Embodiment 7
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1.5h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 10.5h under the condition of ice bath, immobilized enzyme efficiency is 47.4%, the enzyme rate of recovery alive is 65.8%, and the charge capacity of this carrier is 119mg enzyme/g carrier.
Embodiment 8
Be dispersed in the Tris-HCL buffered soln of 15mL pH 8.5 by 40mg nano ferriferrous oxide, add 45mg dopamine hydrochloride, regulate pH to 8.5 with 100mM sodium hydroxide, at room temperature, shake 1.5h under speed 200rpm, after washing, drying obtains solid support material.Getting solid support material 8.3mg joins in the phosphoric acid buffer of pH=8, and then add lipase from Aspergillus Niger enzyme liquid 1ml, fix 18h under the condition of ice bath, immobilized enzyme efficiency is 65.3%, and the enzyme rate of recovery alive is 39.2%, and the charge capacity of this carrier is 71mg enzyme/g carrier.
Claims (10)
1. an enzyme immobilization method, is characterized in that, comprises the steps:
(1) ferriferrous oxide nano-particle is dispersed in the buffered soln of Tris-HCL, is configured to substrate solution;
(2) add dopamine hydrochloride by the substrate solution of gained in (1), adjust pH to be 8.0 to 9.5, at room temperature shake 1 ~ 90min, then repeatedly use deionized water rinsing, dry;
(3) material (2) obtained carries out concussion immobilization with enzyme in phosphoric acid buffer, and immobilization temperature is 0 ~ 4 DEG C, immobilization time 1 ~ 12h;
(4) by after the immobilized enzyme lyophilize of gained in (3).
2. method according to claim 1, is characterized in that, described in step (2), pH is 8.5, and in step (3), the pH of phosphoric acid buffer is 8.0.
3. method according to claim 1, is characterized in that, Iron dichloride tetrahydrate and ferric chloride hexahydrate are obtained by ammoniacal liquor co-precipitation by the ferriferrous oxide nano-particle in described step (1).
4. the method according to claim 1 or 2 or 3, is characterized in that, the mass ratio of described ferriferrous oxide nano-particle and Dopamine HCL is 1:(0.5 ~ 5).
5. the method according to claim 1 or 2 or 3, is characterized in that, in described step (2), (3), concussion speed is 100 ~ 500rpm.
6. the method according to claim 1 or 2 or 3, is characterized in that, the enzyme in described step (2) is lipase or proteolytic enzyme.
7. method according to claim 5, is characterized in that, described lipase is lipase from Aspergillus Niger, the fermented liquid that lipase from Aspergillus Niger is lipase from Aspergillus Niger enzyme powder, born of the same parents contain lipase from Aspergillus Niger after producing lipase from Aspergillus Niger or cell wall breaking outward.
8. according to zymin prepared by claim 1 ~ 6 any one method.
9. zymin according to claim 6, is characterized in that, the optimal temperature of this zymin is 20 ~ 60 DEG C, and pH is 6.5 ~ 9.0.
10. zymin according to claim 9, is characterized in that, the optimal temperature of this zymin is 35 ~ 50 DEG C, and pH is 7.0 ~ 8.0.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105551704A (en) * | 2015-12-09 | 2016-05-04 | 江苏大学 | Preparation and application of dopamine functional magnetic nano-carrier |
| CN106282152A (en) * | 2016-08-08 | 2017-01-04 | 华南理工大学 | A kind of immobilized enzyme and the method being acylated by its catalysis dihydromyricetin |
| CN107973919A (en) * | 2017-11-02 | 2018-05-01 | 天津大学 | A kind of preparation method of pickering emulsion amine stabilized based on DOPA and its immobilised enzymes application |
| CN108273059A (en) * | 2018-02-27 | 2018-07-13 | 山东大学 | A kind of preparation method and application of photo-thermal and the composite nano materials of active oxygen combination therapy tumour |
| CN110004138A (en) * | 2019-04-29 | 2019-07-12 | 中国检验检疫科学研究院 | Immobilised enzymes and preparation method thereof |
| CN110055242A (en) * | 2019-04-29 | 2019-07-26 | 中国检验检疫科学研究院 | Biodiesel and preparation method thereof |
| CN110272895A (en) * | 2019-07-08 | 2019-09-24 | 中国科学院过程工程研究所 | A kind of medium immobilised enzymes and its preparation method and application |
| CN110373408A (en) * | 2019-08-12 | 2019-10-25 | 山东星光首创生物科技有限公司 | One kind is with poly-dopamine-magnetic Fe3O4The method of nanoparticle immobilization D-Psicose 3- epimerase |
| CN111330463A (en) * | 2020-03-04 | 2020-06-26 | 中国科学院过程工程研究所 | Catalytic membrane and preparation method and application thereof |
| CN111876406A (en) * | 2020-06-18 | 2020-11-03 | 南京师范大学 | Magnetic nanoparticle-lipase-metal organic framework composite catalytic material and preparation method and application thereof |
| CN112029758A (en) * | 2020-08-12 | 2020-12-04 | 华南理工大学 | Multi-enzyme immobilization material and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101177678A (en) * | 2006-11-09 | 2008-05-14 | 中国科学院大连化学物理研究所 | Magnetic nano particle enzyme immobilization as well as preparation method and uses thereof |
| CN103232991A (en) * | 2013-04-11 | 2013-08-07 | 复旦大学 | A synthesis method for a dopamine-modified magnetic mesoporous silica material and applications thereof |
| CN103887030A (en) * | 2014-04-04 | 2014-06-25 | 华东理工大学 | Magnetic affinity nanometer particle used for purifying and immobilizing histidine tag protein, and preparation method and application thereof |
-
2015
- 2015-05-15 CN CN201510250417.8A patent/CN104894094A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101177678A (en) * | 2006-11-09 | 2008-05-14 | 中国科学院大连化学物理研究所 | Magnetic nano particle enzyme immobilization as well as preparation method and uses thereof |
| CN103232991A (en) * | 2013-04-11 | 2013-08-07 | 复旦大学 | A synthesis method for a dopamine-modified magnetic mesoporous silica material and applications thereof |
| CN103887030A (en) * | 2014-04-04 | 2014-06-25 | 华东理工大学 | Magnetic affinity nanometer particle used for purifying and immobilizing histidine tag protein, and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 张玉玮等: "基于仿生聚多巴胺膜和纳米金的酶固定化平台的构建", 《化学学报》 * |
| 谢槟等: "基于多巴胺自聚合及肝素固定改善钛的血液相容性", 《高等学校化学学报》 * |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105551704A (en) * | 2015-12-09 | 2016-05-04 | 江苏大学 | Preparation and application of dopamine functional magnetic nano-carrier |
| CN106282152A (en) * | 2016-08-08 | 2017-01-04 | 华南理工大学 | A kind of immobilized enzyme and the method being acylated by its catalysis dihydromyricetin |
| CN107973919A (en) * | 2017-11-02 | 2018-05-01 | 天津大学 | A kind of preparation method of pickering emulsion amine stabilized based on DOPA and its immobilised enzymes application |
| CN108273059B (en) * | 2018-02-27 | 2021-02-19 | 山东大学 | Preparation method and application of composite nano material for treating tumors by combining photo-thermal treatment and active oxygen treatment |
| CN108273059A (en) * | 2018-02-27 | 2018-07-13 | 山东大学 | A kind of preparation method and application of photo-thermal and the composite nano materials of active oxygen combination therapy tumour |
| CN110004138A (en) * | 2019-04-29 | 2019-07-12 | 中国检验检疫科学研究院 | Immobilised enzymes and preparation method thereof |
| CN110055242A (en) * | 2019-04-29 | 2019-07-26 | 中国检验检疫科学研究院 | Biodiesel and preparation method thereof |
| CN110272895A (en) * | 2019-07-08 | 2019-09-24 | 中国科学院过程工程研究所 | A kind of medium immobilised enzymes and its preparation method and application |
| CN110373408A (en) * | 2019-08-12 | 2019-10-25 | 山东星光首创生物科技有限公司 | One kind is with poly-dopamine-magnetic Fe3O4The method of nanoparticle immobilization D-Psicose 3- epimerase |
| CN110373408B (en) * | 2019-08-12 | 2021-06-25 | 山东星光首创生物科技有限公司 | Method for immobilizing D-psicose 3-epimerase by polydopamine-magnetic Fe3O4 nanoparticles |
| CN111330463A (en) * | 2020-03-04 | 2020-06-26 | 中国科学院过程工程研究所 | Catalytic membrane and preparation method and application thereof |
| CN111330463B (en) * | 2020-03-04 | 2021-06-08 | 中国科学院过程工程研究所 | Catalytic membrane and preparation method and application thereof |
| CN111876406A (en) * | 2020-06-18 | 2020-11-03 | 南京师范大学 | Magnetic nanoparticle-lipase-metal organic framework composite catalytic material and preparation method and application thereof |
| CN111876406B (en) * | 2020-06-18 | 2022-05-03 | 南京师范大学 | Magnetic nanoparticle-lipase-metal organic framework composite catalytic material and preparation method and application thereof |
| CN112029758A (en) * | 2020-08-12 | 2020-12-04 | 华南理工大学 | Multi-enzyme immobilization material and preparation method and application thereof |
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Application publication date: 20150909 |