CN113801875A - Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material - Google Patents
Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material Download PDFInfo
- Publication number
- CN113801875A CN113801875A CN202110536746.4A CN202110536746A CN113801875A CN 113801875 A CN113801875 A CN 113801875A CN 202110536746 A CN202110536746 A CN 202110536746A CN 113801875 A CN113801875 A CN 113801875A
- Authority
- CN
- China
- Prior art keywords
- enzyme
- carbon material
- immobilized
- functionalized
- functionalized carbon
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 39
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 39
- 239000003575 carbonaceous material Substances 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 19
- 230000003100 immobilizing effect Effects 0.000 title claims description 6
- 108010093096 Immobilized Enzymes Proteins 0.000 claims abstract description 18
- 238000001179 sorption measurement Methods 0.000 claims abstract description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 10
- 239000002041 carbon nanotube Substances 0.000 claims description 7
- 229910021393 carbon nanotube Inorganic materials 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 2
- 229920001308 poly(aminoacid) Polymers 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000000346 sugar Nutrition 0.000 claims description 2
- 150000008163 sugars Chemical class 0.000 claims description 2
- 229920000049 Carbon (fiber) Polymers 0.000 claims 1
- 239000004917 carbon fiber Substances 0.000 claims 1
- 229910021389 graphene Inorganic materials 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 9
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 230000002708 enhancing effect Effects 0.000 abstract description 4
- 230000000379 polymerizing effect Effects 0.000 abstract description 3
- 230000007547 defect Effects 0.000 abstract description 2
- 229940088598 enzyme Drugs 0.000 description 25
- 102000016943 Muramidase Human genes 0.000 description 10
- 108010014251 Muramidase Proteins 0.000 description 10
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 10
- 229960000274 lysozyme Drugs 0.000 description 10
- 239000004325 lysozyme Substances 0.000 description 10
- 235000010335 lysozyme Nutrition 0.000 description 10
- 108010039918 Polylysine Proteins 0.000 description 7
- 229920000656 polylysine Polymers 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000006555 catalytic reaction Methods 0.000 description 3
- 238000007306 functionalization reaction Methods 0.000 description 3
- 108020005199 Dehydrogenases Proteins 0.000 description 2
- 108010015776 Glucose oxidase Proteins 0.000 description 2
- 239000004366 Glucose oxidase Substances 0.000 description 2
- 102000004157 Hydrolases Human genes 0.000 description 2
- 108090000604 Hydrolases Proteins 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000012876 carrier material Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 229940116332 glucose oxidase Drugs 0.000 description 2
- 235000019420 glucose oxidase Nutrition 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 239000002198 insoluble material Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000003278 mimic effect Effects 0.000 description 2
- 229940072417 peroxidase Drugs 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000011942 biocatalyst Substances 0.000 description 1
- 238000010364 biochemical engineering Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- -1 lywallzyme Proteins 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
Landscapes
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The invention discloses an immobilized enzyme method based on a polymerized biomolecule functionalized carbon material. In order to overcome the defects in the prior immobilized enzyme technology, a carbon material is functionalized by polymerizing biological molecules, and the immobilized enzyme is obtained by taking the functionalized carbon material as a carrier of the immobilized enzyme in an adsorption mode. The invention not only retains the strong adsorption force of the carbon material, but also protects the protein structure of the enzyme, and plays a role in maintaining and enhancing the enzyme activity. The method has the advantages of simple operation, simple procedure, high specific activity and the like.
Description
Technical Field
The invention relates to the technical field of immobilized enzymes, in particular to an immobilized enzyme method based on a polymeric biomolecule functionalized carbon material.
Background
The enzyme is a biological macromolecular protein, can participate in various catalytic reactions under the conditions of normal temperature and normal pressure, and is a high-efficiency catalyst. The biocatalyst has the characteristics of low requirement on catalysis conditions, specificity, fast catalysis process and the like. Therefore, enzymes have been widely used in various industries and fields, such as environmental monitoring, clinical diagnosis, food processing, and biochemical engineering. However, the stability of the enzyme in solution is poor and it is difficult to use it repeatedly and for a long period of time. The immobilized enzyme technology is characterized in that some materials with excellent physicochemical properties are selected, the materials need to have larger pore size or have strong adsorption force, and the like, and finally, the enzyme is tightly bound in the insoluble materials or on the surface of the insoluble materials to realize immobilization. With the development of the immobilized enzyme technology, the advantages of the immobilized enzyme are reflected. Compared with solution enzyme, the stability and the tolerance to environmental change of the enzyme are improved, and the application range of the enzyme is wider.
The traditional methods for immobilizing enzymes include adsorption, covalent bonding, embedding and the like. Carbon materials are often studied as carrier materials for immobilized enzymes due to their strong adsorption force, but the carbon materials often cause changes in the protein structure of the enzymes, resulting in reduction or even inactivation of the enzyme activity due to their too strong adsorption force. The carbon material is functionalized by innovatively using different polymeric biomolecules based on an immobilized enzyme technology, and the carbon material is used as a carrier material of the immobilized enzyme, so that the strong adsorption capacity of the carbon material is reserved, the protein structure of the enzyme can be protected, and the effect of maintaining and enhancing the activity of the enzyme is achieved.
Disclosure of Invention
In order to overcome the defects in the prior immobilized enzyme technology, a carbon material is functionalized by polymerizing biological molecules, and the immobilized enzyme is obtained by taking the functionalized carbon material as a carrier of the immobilized enzyme in an adsorption mode. The invention not only retains the strong adsorption force of the carbon material, but also protects the protein structure of the enzyme, and plays a role in maintaining and enhancing the enzyme activity. The method has the advantages of simple operation, simple procedure, high specific activity and the like.
Further, the polymeric biomolecules include, but are not limited to, polyamino acids, DNA, RNA, sugars, starch, biolipids.
Further, the functionalization method refers to polymerizing biological molecules as a functionalization agent, and functionalizing the surface of the carbon material in an adsorption or covalent linkage manner, wherein the surface of the carbon material is covered or partially covered by the functionalization agent;
further, the enzyme includes biological enzymes or artificial mimic enzymes, including but not limited to various oxidoreductases, hydrolases, dehydrogenases, etc., including but not limited to lysozyme, lywallzyme, glucose oxidase, peroxidase. List of partial enzymes.
Further, the immobilized enzyme includes immobilized biological enzyme or artificial mimic enzyme, including but not limited to various oxidoreductases, hydrolases, dehydrogenases, etc., including but not limited to lysozyme, glucose oxidase, peroxidase. List of partial enzymes.
Further, the method of immobilizing the enzyme means that the enzyme is immobilized on the functionalized carbon material in an adsorption manner.
Drawings
FIG. 1 is a graph showing the results of lysozyme immobilized by polylysine-functionalized carbon nanotubes according to the present invention.
Detailed Description
Specific activity of lysozyme immobilized on polylysine functionalized carbon material was compared, wherein polylysine was epsilon-polylysine, working buffer solution: pH 7.4,10 mmol/l, 0.9% sodium chloride. Specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
When the lysozyme is directly immobilized by using the carbon nano tube, the secondary structure of the lysozyme is damaged, and the inactivation of the lysozyme is caused. The carbon nano tube is functionalized by using polylysine with bactericidal activity, and the lysozyme is immobilized by the polylysine functionalized carbon nano tube in an adsorption mode. Polylysine not only retains the strong adsorption force of the carbon nano tube, but also changes the microstructure of the surface of the carbon nano tube and protects the protein structure of enzyme, and the polylysine not only shows bactericidal activity, but also plays a role in synergy and enhancing the bactericidal effect of lysozyme, so that the activity of the lysozyme is obviously enhanced.
Claims (6)
1. A method for immobilizing an enzyme based on a polymeric biomolecule functionalized carbon material is characterized by comprising the following specific steps:
(1) the method for immobilizing the enzyme based on the polymeric biomolecule functionalized carbon material comprises the steps of functionalizing the surface of the carbon material by adsorption or covalent linkage by taking the polymeric biomolecule as a functionalizing agent, wherein the surface of the carbon material is covered or partially covered by the functionalizing agent;
(2) the enzyme is immobilized on the functionalized carbon material described in (1) by adsorption.
2. The method of claim 1, wherein the carbon material includes, but is not limited to, carbon nanotubes, graphene, porous carbon, activated carbon, carbon fibers.
3. The method of claim 1, wherein said polymeric biomolecules include, but are not limited to, polyamino acids, DNA, RNA, sugars, starch, biolipids.
4. The method of claim 1, wherein the enzyme comprises a biological enzyme or an artificial mimetic enzyme.
5. The method of claim 1, wherein the immobilized enzyme comprises an immobilized biological enzyme or an artificial mimetic enzyme.
6. The method of claim 1, wherein the immobilized enzyme is a single enzyme or a combination of enzymes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110536746.4A CN113801875A (en) | 2021-05-18 | 2021-05-18 | Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110536746.4A CN113801875A (en) | 2021-05-18 | 2021-05-18 | Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113801875A true CN113801875A (en) | 2021-12-17 |
Family
ID=78942404
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110536746.4A Pending CN113801875A (en) | 2021-05-18 | 2021-05-18 | Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113801875A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421762A (en) * | 2012-05-22 | 2013-12-04 | 北京化工大学 | Immobilized enzyme and preparation method thereof |
| CN104132982A (en) * | 2014-07-25 | 2014-11-05 | 山东省科学院生物研究所 | Dehydrogenase electrode and preparation method thereof |
-
2021
- 2021-05-18 CN CN202110536746.4A patent/CN113801875A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421762A (en) * | 2012-05-22 | 2013-12-04 | 北京化工大学 | Immobilized enzyme and preparation method thereof |
| CN104132982A (en) * | 2014-07-25 | 2014-11-05 | 山东省科学院生物研究所 | Dehydrogenase electrode and preparation method thereof |
Non-Patent Citations (7)
| Title |
|---|
| AVIRU KUMAR BASU等: "Poly-L-lysine functionalised MWCNT-rGO nanosheets based 3-dhybrid structure for femtomolar level cholesterol detection using cantilever based sensing platform", SCIENTIFIC REPORTS * |
| DANIEL W.HORN等: "Lysozyme dispersed single-walled carbon nanotubes: interaction and activity", J.PHYS CHEM C * |
| DANIELE MERLI 等: "Increasing the antibacterial effect of lysozyme by immobilization on multi-walled carbon nanotubes", JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY * |
| LENIHAN J.S.等: "Protein immobilization on carbon nanotubes through a molecular adapter", J. NANOSCI.NANOTECH * |
| YUANJIAN ZHANG 等: "Poly-L-lysine functionalization of single-walled carbon nanotubes", J. PHYS. CHEM.B * |
| YUANXING CAI等: "Preparation of chitosan modified magnetic carbon nanotubes and application in immobilized enzymes", COMPOSITE INTERFACE * |
| 王超: "氨基酸调节聚天冬氨酸功能化碳纳米管及对溶菌酶吸附", 中国优秀硕士学位论文全文数据库 工程科技I辑 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Min et al. | Recent progress in nanobiocatalysis for enzyme immobilization and its application | |
| Sirisha et al. | Enzyme immobilization: an overview on methods, support material, and applications of immobilized enzymes | |
| Thangaraj et al. | Immobilization of lipases–a review. Part II: carrier materials | |
| Kumar et al. | Immobilization of soybean (Glycine max) urease on alginate and chitosan beads showing improved stability: Analytical applications | |
| US4102746A (en) | Immobilized proteins | |
| Kujawa et al. | Highly effective enzymes immobilization on ceramics: Requirements for supports and enzymes | |
| El‐Zahab et al. | Enabling multienzyme biocatalysis using nanoporous materials | |
| US4169014A (en) | Method of immobilizing proteinaceous substances | |
| Elnashar et al. | Lactose hydrolysis by β-galactosidase covalently immobilized to thermally stable biopolymers | |
| Sneha et al. | Enzyme immobilization methods and applications in the food industry | |
| Nwagu et al. | Immobilization of a saccharifying raw starch hydrolyzing enzyme on functionalized and non-functionalized sepa beads | |
| Wang et al. | Improving pullulanase catalysis via reversible immobilization on modified Fe 3 O 4@ polydopamine nanoparticles | |
| Ayhan et al. | Highly biocompatible enzyme aggregates crosslinked by L-lysine | |
| CN109266639B (en) | Dual immobilized enzyme and preparation method and application thereof | |
| Carneiro-da-Cunha et al. | Lipase immobilisation on to polymeric membranes | |
| Anwar et al. | Smart chemistry and applied perceptions of enzyme-coupled nano-engineered assemblies to meet future biocatalytic challenges | |
| Bautista et al. | Properties of a glucose oxidase covalently immobilized on amorphous AlPO4 support | |
| Spasojević et al. | The enzyme immobilization: carriers and immobilization methods | |
| KR102276595B1 (en) | Enzyme-carrier composite | |
| CN113801875A (en) | Method for immobilizing enzyme based on polymerized biomolecule functionalized carbon material | |
| Wang et al. | Influence of silica-derived nano-supporters on cellobiase after immobilization | |
| Vasileva et al. | Study on the behaviour of glucose oxidase immobilized on microfiltration polyamide membrane | |
| CN113481192A (en) | Method for immobilizing enzyme based on metal oxide and metal hydroxide functionalized carbon material | |
| CN114045327B (en) | Biological sensing film and preparation method and application thereof | |
| CN113430191B (en) | Immobilized enzyme method based on organic polymer functionalized carbon material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211217 |