Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

PVA membrane immobilized enzyme and preparation method thereof

A technology for immobilizing enzymes and porous membranes, applied in the directions of immobilizing enzymes, biochemical equipment and methods, enzymes, etc., can solve the problems of complex process of immobilizing enzymes in porous membranes, achieve good immobilization effect, simple process, and improve load amount of effect

Inactive Publication Date: 2021-09-03
ASYMCHEM LIFE SCI TIANJIN
View PDF11 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The main purpose of the present invention is to provide a kind of PVA membrane immobilized enzyme and preparation method thereof, to solve the complex problem of adopting porous membrane immobilized enzyme process in the prior art

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • PVA membrane immobilized enzyme and preparation method thereof
  • PVA membrane immobilized enzyme and preparation method thereof
  • PVA membrane immobilized enzyme and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0070] In another typical embodiment of the present application, a method for preparing any of the above PVA membrane-immobilized enzymes is provided, the preparation method comprising: step S1, mixing the raw materials containing the enzyme and the PVA solution for a predetermined time to obtain a mixed system ; Step S2, adding the mixed system to the mold and drying the mixed system to obtain membrane-embedded enzymes, the mold is a three-dimensional structured mold to form a three-dimensional structured PVA porous membrane; and step S3, using phosphate buffer to After the membrane-embedded enzyme is soaked and washed, the PVA membrane-immobilized enzyme is obtained, and the pH value of the mixed system is preferably 6.0-6.5.

[0071] The preparation method of the present application adopts mixing, drying and post-treatment processes to form the PVA membrane immobilized enzyme, the process is simple, easy to operate, and does not need to use glutaraldehyde, amino, carboxyl, e...

Embodiment 1

[0108] Immobilization of transaminase TA-CV CLEA (cross-linked enzyme) by PVA membrane embedding:

[0109] Preparation of PVA I solution: 50mL 10% (w / v) PVA (200KDa) was mixed with 30mL 10% (w / v) acetic acid, 50% (v / v) methanol, 10% (w / v) sulfuric acid.

[0110] PVA membrane encapsulation: adjust the pH of the PVA I solution to 6.0, and take out 20mL of the solution, mix it with the suspension of TA-CV cross-linking enzyme aggregates (CLEA) (the suspension of TA-CV cross-linking enzyme aggregates CLEA 0.5 g of TA-CV cross-linking enzyme in 2 ml of 0.1 M phosphate buffer (PB) pH 7.0, each ml containing 2 mg of PLP (pyrrolealdehyde phosphate)) was stirred for 20 min to form a mixed system. Pour it into a 3D porous silica gel template. The bullet holes in each template are square, with a volume of about 0.1 to 0.2 cubic centimeters and a bullet hole surface area of ​​about 2 to 5 square centimeters. Dry at 37°C to obtain membrane-embedded enzyme. Soak the membrane-embedded enzy...

Embodiment 2

[0129] Immobilization of transaminase TA-CV CLEA by PVA membrane embedding

[0130] Preparation of PVA II solution: 12%-15% (w / v) PVA aqueous solution.

[0131] PVA film embedding: Take 30mL PVA solution, add 3 grams of CLEA wet particles, and 50mg of PLP, stir well to form a mixed system, pour it into a 3D porous silica gel template, and the bullet holes in each template are circular, with a volume of about 0.15~ 0.2 cubic centimeter, the bullet hole surface area is about 3-5 square centimeters, and dried at 37°C to obtain membrane-embedded enzyme. Soak the membrane-embedded enzyme in 0.1MPB 7.0 buffer overnight, remove the membrane-embedded enzyme from the buffer, and then wash the membrane-embedded enzyme twice with 0.1M PB7.0 buffer to obtain the example 2 PVA membrane immobilized enzymes.

[0132] Before adding CLEA, a certain amount of PEG 400 ~ PEG6000 were dissolved in PVA II solution for parallel experiments.

[0133] Carry out conversion and stability test accordi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a PVA membrane immobilized enzyme and a preparation method thereof. The PVAfilm immobilized enzyme comprises a PVA porous membrane and an enzyme embedded on the PVA porous membrane, the PVA porous membrane is a three-dimensional structured PVA porous membrane, and the enzyme is selected from any one of transaminase, D-lactic dehydrogenase, cyclohexanone monooxygenase, ketoreductase, enoyl reductase, nitrilase, ammonia lyase, amino acid dehydrogenase, imine reductase, alcohol dehydrogenase, ammonium formate dehydrogenase, glucose 1-dehydrogenase and mutants thereof. The three-dimensional structured PVA porous membrane is adopted as a carrier to immobilize enzyme in an embedding manner, the embedding immobilization process is simple, the conditions are mild, the specific surface area is large, and the enzyme is relatively stable after being embedded and immobilized in the PVA porous membrane and is not easy to leach in the use process; and the porous structure of the PVA porous membrane can better transfer reactants and products, is suitable for being used in continuous flow biochemical catalysis, and has wide adaptability to enzymes.

Description

[0001] This application is a divisional application based on a patent application with an application date of July 16, 2020, an application number of 202010683476.5, and an invention titled "PVA membrane immobilized enzyme and its preparation method". technical field [0002] The invention relates to the technical field of enzyme immobilization, in particular to a PVA film immobilized enzyme and a preparation method thereof. Background technique [0003] Biocatalysis is becoming an integral part of molecular manufacturing programs for chemicals, intermediates, fine chemicals and ultimately pharmaceuticals. However, as process demands continue to expand, the efficiency and economy of enzyme applications becomes inevitable. Therefore, it is not only necessary to enhance enzyme activity, specificity and productivity, but also to improve shelf life and recyclability, especially to facilitate the economic viability of commercial-scale applications. [0004] The Enzyme Immobiliza...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/084C12N11/089
CPCC12N11/084C12N11/089C12N11/04
Inventor 洪浩詹姆斯·盖吉肖毅张娜罗杰斯卡·维亚撒·威廉姆斯崔瑜霞赵佳东高妍妍
Owner ASYMCHEM LIFE SCI TIANJIN
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com