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

Preparation method for partially inactivated chymosin

A technology of rennet and preparation method, which is applied in the field of protein renaturation, can solve the problems of inactivity, etc., and achieve the effect of simple operation, obvious renaturation effect and good repeatability

Active Publication Date: 2017-12-15
SHANDONG POLYTECHNIC
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the expression system itself or the treatment of Guo Chen in the experiment, these proteins mostly exist in an inactive form and need to be refolded

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
  • Preparation method for partially inactivated chymosin
  • Preparation method for partially inactivated chymosin
  • Preparation method for partially inactivated chymosin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The chymosin solution containing 0, 0.50, 0.75, 1.00, 1.25 and 1.5mmol / L ectoine was placed at 69°C for 0, 1min, 2min, 3min, 4min, 5min, 6min, 7min, 8min, respectively. 9min, 10min, obtain partially inactivated chymosin; figure 1 It is the time-course curve of the change of chymosin activity under different concentrations of ectoine. Depend on figure 1 It can be seen that after adding ectoine, the thermal stability of chymosin in each group is reduced to varying degrees. In each group containing ectoine, when ectoine is 0.75mmol / L, the heat treatment time for enzyme activity can be measured At the longest time, 9% of the relative activity remained after 7 minutes of treatment; for other concentrations, the relative activity dropped to about 10% at 4-6 minutes.

Embodiment 2

[0040] The chymosin solution containing 0.75mmol / L ectoine was placed at temperatures of 61, 63, 65, 67, 68, 69 and 70°C for heat inactivation treatment for different times to obtain partially inactivated chymosin; said figure 2 It is the time-course curve of the change of chymosin activity under the same ectoine concentration.

[0041] Refolding experiment

[0042] 1. Effect of heat treatment time on refolding of chymosin

Embodiment 3

[0044] The partially inactivated rennet in Example 1 was rapidly cooled to 4° C. and kept in a refrigerator for static storage. Samples were taken every 6 hours from 0 to 42 hours, and the water bath was preheated to 35°C to measure the change of the residual activity of chymosin. image 3 ,Depend on image 3It can be seen that with the prolongation of renaturation time, the activity of chymosin increased. The residual enzyme activity of the non-renaturation control group was 61.5% after incubation for 1 minute; after 42 hours of renaturation, the residual enzyme activity rose to 83.3%. In addition, the heat treatment time at which rennet activity could be detected was also significantly prolonged. In the control group, the enzyme activity could not be detected after 6 minutes of heat treatment. After 12 minutes of heat treatment, 8.8% of the remaining enzyme activity could be detected in the experimental group after renaturation for 36 hours and 42 hours.

[0045] 2. Effect...

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 belongs to the technical field of protein renaturation and especially relates to a preparation method for partially inactivated chymosin. The preparation method comprises the following steps: (1) placing a chymosin solution containing ectoine under the temperature at 61-70 DEG C and treating for 1-20min, thereby acquiring the partially inactivated chymosin, and (2) quickly cooling the partially inactivated chymosin prepared in the step (1) to 4 DEG C, standing by and refrigerating for 0-42h, thereby acquiring the chymosin after renaturation. The method is different from the treating method for high concentration urea, and the like; a heat inactivation method is adopted for preparing the partially inactivated chymosin; the operation is simple and easy, the repeatability is high and the state of the partially inactivated chymosin is more approximate to the heat inactivation state of enzyme; the inactivated chymosin prepared according to the method has the characteristics of obvious renaturation effect, high enzyme activity after renaturation, and the like.

Description

technical field [0001] The invention belongs to the technical field of protein refolding, in particular to a preparation method of partially inactivated chymosin. Background technique [0002] Rennet is an essential enzyme preparation for making cheese and casein, and it can hydrolyze Phe of κ-casein in milk 105 -Met 106 Peptide bonds make the milk coagulate, and after a series of processing, it is made into cheese. In addition, rennet also has a clinical effect in treating dyspepsia in children. Like other enzyme preparations, rennet may be denatured and inactivated by high heat, freeze-thaw, radiation, organic solvents, etc. during production and application, resulting in loss of enzyme activity. Cheese processing has strict requirements on the amount of rennet added, and as a pharmaceutical preparation, it is also required that rennet can maintain high stability. Methods such as immobilization, chemical modification, and protein engineering can all improve the thermal...

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): C12N9/50
CPCC12N9/50C12Y304/23004
Inventor 武彬
Owner SHANDONG POLYTECHNIC
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