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

Self-induction-based method for biomimetic titanification of immobilized protease

An immobilized protease self-induction technology, applied in the field of immobilized enzyme preparation, achieves the effects of mild conditions, simple immobilization process, and reduced energy consumption

Inactive Publication Date: 2017-09-15
GUILIN UNIVERSITY OF TECHNOLOGY
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the defects of traditional immobilization methods, and provide a method based on self-induced biomimetic titanation immobilized papain

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
  • Self-induction-based method for biomimetic titanification of immobilized protease
  • Self-induction-based method for biomimetic titanification of immobilized protease
  • Self-induction-based method for biomimetic titanification of immobilized protease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] At room temperature, add 7.5mL 0.25mol / L Ti-BALDH solution to the immobilization reactor, and then add 22.5mL to the immobilization reactor to prepare the concentration of 18mg with pH 7.5 and 0.06mol / L phosphate buffer / mL free papain solution, so that the volume ratio of Ti-BALDH solution and papain solution is 1:3; gently stir for 5min, 5000rpm refrigerated centrifugation for 10min, and collect the precipitate. The precipitate was washed with distilled water for 3 times, the washing liquid was collected, the supernatant was combined, and then the precipitate was freeze-dried for 24 hours. The recovery rate of the obtained immobilized enzyme activity was 51.76%, and the encapsulation rate of papain was 78.67%.

Embodiment 2

[0022] Take 20 mg of the immobilized papain and free protease obtained in Example 1 and incubate in a 55°C water bath, take a sample every 1h and add phosphate buffer at an optimal pH, then shake and react with casein in a 37°C water bath for 10 minutes, The residual enzyme activity was measured, and the enzyme activity of the sample without heat preservation treatment was taken as 100%, and the others were converted into relative enzyme activities to investigate the temperature stability of the immobilized papain. attached figure 1 is the relative enzyme activity of free and immobilized papain incubated at pH 7.5 with 55°C water bath for 5h. It can be seen from the figure that the free enzyme can maintain more than 80% of the relative enzyme activity after incubation in a 55°C water bath for 1 hour, but as the incubation time prolongs, the relative activity of free papain decreases rapidly, and only maintains the initial enzyme activity after 5 hours of incubation 39 percent...

Embodiment 3

[0024] For the immobilized papain obtained in Example 1, after the sample is ground into fine pieces, potassium bromide is pressed into tablets, and its infrared spectrum is measured by a Fourier transform infrared spectrometer, and TiO is catalyzed with papain and ammonia water. 2 Infrared spectra for comparison, attached figure 2 shown. It can be seen from the figure that, compared with spectral line B, spectral line A has multiple absorption peaks of characteristic groups. Among them, 1372cm -1 It is the Ti-O-Ti antisymmetric stretching vibration absorption peak; 1116cm -1 It is the Ti-O-H stretching vibration absorption peak; 668cm -1 It is the Ti-O-Ti symmetrical stretching vibration absorption peak; 537cm -1 It is the bending vibration absorption peak of Ti-O-Ti, which further proves that TiO 2 generation. In addition, 1652 and 1516cm -1 The absorption peak at 2872cm is attributed to the vibrational absorption of the amide group (amide I band and II band) in the ...

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 self-induction-based method for biomimetic titanification of immobilized protease. Immobilization of the protease is achieved in the induced biomimetic titanification process by using the inductive effect of papain on biomimetic titanification of a titanium precursor dihydroxy bis(ammonium lactate) titanium to obtain the immobilized protease employing titanium oxide as a carrier. The defects that a traditional immobilization method is complicated in immobilization process, the immobilized protease is poor in biocompatibility of an enzyme and the carrier, relatively weak in bonding action with the carrier, poor in stability and short in service life, and the immobilization reaction is relatively violent and easily causes enzyme inactivation and activity reduction are overcome, and the method has the advantages of being simple in immobilization process, mild in condition, high in efficiency, good in enzyme recovery rate of the immobilized protease and good in embedding rate.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzyme preparation, in particular to a method based on self-induced biomimetic titanation immobilized protease. Background technique [0002] The chemical essence of enzyme is protein, and its catalysis has the characteristics of high selectivity, high catalytic activity, mild reaction conditions, environmental protection and no pollution. However, the enzyme in the free state has poor stability to heat, strong acid, strong alkali, high ionic strength, organic solvents, etc., and is easily inactivated, and after the reaction, it is mixed with catalytic products and other substances, making it difficult to purify and cannot be reused. In order to overcome these problems, enzyme immobilization technology came into being. [0003] Traditional immobilization methods include: adsorption method, cross-linking method, covalent bonding method, embedding method, etc., adsorption method, cross-linking ...

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): C12N11/02
CPCC12N9/63C12N11/02C12Y304/22002
Inventor 李海云韩文余艳葵阮贵华李子院
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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