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

Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles

A technology of fe3o4sio2-nh2 and nano microspheres, which is applied in the field of marine microorganisms, can solve the problems of limiting the utilization of marine lipase, difficulties in the separation and purification of lipase, and poor stability of free enzymes, and achieves low production costs, high mechanical strength, and enzyme production. The effect of temperature stabilization

Inactive Publication Date: 2013-10-02
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The emergence of immobilized enzymes has solved many limitations in the practical application of free enzymes, such as poor stability of free enzymes, difficulties in the separation and purification of lipases, etc., which limit the utilization of marine lipases

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
  • Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles
  • Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles
  • Method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] 10mg magnetic Fe 3 O 4 SiO 2 -NH 2 The nanospheres were placed in a 10ml vial, and then 2ml of lipase solution with an enzyme protein concentration of 0.125 mg / ml and an immobilization system of pH 8.0 was added to the vial, and the immobilization reaction temperature was 30°C on a shaker. Carry out the immobilization reaction for 10 hours, and then perform magnetic separation. Collect the supernatant and use it to determine the protein content. After washing the immobilized enzyme several times, until there is no protein content absorbed in the supernatant, the enzyme is used to determine the activity of the immobilized enzyme. The immobilized marine lipase Bohai Sea-9145 was obtained, the enzyme activity recovery rate was 87.7%, and the protein adsorption rate per unit area of ​​the carrier was 23.7 mg / g.

Embodiment 2

[0092] 10mg magnetic Fe 3 O 4 SiO 2 -NH 2 Place the nanospheres in a 10ml vial, and then add 2ml of lipase solution with an enzyme protein concentration of 0.150 mg / ml, immobilization system pH of 7.0, and an immobilization reaction temperature of 25°C in the vial. Carry out the immobilization reaction for 12 hours, and then perform magnetic separation. Collect the supernatant and use it to determine the protein content. After washing the immobilized enzyme several times, until no protein content is absorbed in the supernatant, the enzyme is used to determine the activity of the immobilized enzyme. The immobilized marine lipase Bohai Sea-9145 was obtained, the enzyme activity recovery rate was 86.5%, and the protein adsorption rate per unit area of ​​the carrier was 24.0 mg / g.

Embodiment 3

[0094] 10mg magnetic Fe 3 O 4 SiO 2 -NH 2 Nanospheres were placed in a 10ml vial, and 2ml of lipase solution with a concentration of 0.1mg / ml of enzyme protein and a pH of 7.0 for the immobilization system was added to the vial. The immobilization reaction temperature was 20℃, and the immobilization reaction was carried out on a shaker. After 14 hours, magnetic separation was performed, and the supernatant was collected and used to determine the protein content. After washing the immobilized enzyme several times, until no protein content was absorbed in the supernatant, the enzyme was used to determine the activity of the immobilized enzyme. Lipase Bohai Sea-9145 has an enzyme activity recovery rate of 81.0%, and a protein adsorption rate per unit area of ​​the carrier is 22.0 mg / g.

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 relates to a method for immobilizing marine lipase Bohai Sea-9145 with magnetic Fe3O4@SiO2-NH2 nanoparticles. The method is characterized by comprising the following steps of: putting the magnetic Fe3O4@SiO2-NH2 nanoparticles in a reactor, then adding a marine lipase Bohai Sea-9145 solution with enzyme protein concentration of 0.0375-0.225mg / ml and pH value of 4.0-10.0 to the reactor to implement an immobilization reaction for 4-20 hours at 20-50 DEG C, then carrying out magnetic separation, collecting the supernatant for determining the content of the protein, and washing immobilized marine lipase Bohai Sea-9145 until no protein content in the washing solution is absorbed, thereby obtaining immobilized marine lipase Bohai Sea-9145. The method has the characteristics that the enzyme activity stability promotion effect of the immobilized marine lipase Bohai Sea-9145 is obvious, the enzyme activity recovery rate is as high as 87.7%, the obtained immobilized enzyme has good reutilization property and high mechanical strength and can still keep higher lipase activity after being stored for a long time, and the method is an ideal enzyme immobilization method; and therefore the method is more widely applied to industries, medicines, biochemical analysis and the like.

Description

Technical field [0001] The present invention relates to the field of marine microorganisms, in particular to magnetic Fe 3 O 4 SiO 2 -NH 2 Method for immobilizing marine lipase Bohai Sea-9145 with nano-microspheres. Background technique [0002] Lipase (EC3.1.1.3) is a type of carboxyl hydrolase that mainly catalyzes the hydrolysis of glycerol and long-chain fatty acids inside and outside the cell, and ester synthesis. It is an interfacial enzyme that degrades the substrate to form free fatty acids and glycerol. It belongs to a class of hydrolases (EC3), acting on the ester bond (EC3.1.1) of carboxylic acid esters (EC3.1), and can hydrolyze triglycerides, diglycerides and monoglycerides, and is called triacyl Glycerol acyl hydrolase (EC3.1.1.3). Lipase has a wide range of catalytic reactions and can catalyze a variety of biochemical reactions including hydrolysis, transesterification, alcoholysis, acidolysis, esterification and aminolysis. Lipase can play a role in regioselecti...

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
IPC IPC(8): C12N11/14
Inventor 孙谧徐甲坤盛军郑鸿飞刘均忠琚彩霞
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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