Method for conducting catalytic degradation on isoflavones by using immobilized laccases
A technology for immobilizing laccase and catalyzing degradation, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, immobilized on or in inorganic carriers, and can solve the problems of limited industrial applications, low utilization of free enzymes, and non-recyclability and other problems, to achieve the effect of improving operational stability and cycle times, overcoming easy inactivation and denaturation, and good application potential
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0025] Example 1 Preparation of immobilized laccase
[0026] (1) Preparation of magnetic source particles
[0027] Weigh 10.125 g FeCl respectively 3 ·6H 2 O and 27 g CH 3 COONa was added to 300 mL of ethylene glycol solution, sonicated for 5 min to dissolve, then 7.5 mL of PEG40000 saturated solution was added, magnetically stirred for 15 min, the reaction mixture was transferred to a three-necked flask, and condensed using an electric heating mantle (180˚C) The reaction was refluxed for 8 h, and after cooling to room temperature, the magnetic source microspheres (Fe 3 O 4 ), washed several times with anhydrous ethanol, and dried at 50˚C for later use.
[0028] (2) Preparation of magnetic silica microspheres
[0029] Weigh 1 g of the prepared Fe 3 O 4 , added to 400 mL of 80% ethanol solution, sonicated for 1 h, added 12 mL of concentrated ammonia water to the reaction solution, stirred at room temperature for 15 min, and then added 4 mL of TEOS and continued to stir ...
Embodiment 2
[0035] Example 2 Study on the enzymatic properties of immobilized laccase
[0036] Determination of optimum reaction pH: free enzyme and immobilized enzyme use ABTS as substrate, respectively, react at different pH (2.5, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0) and 40 °C, and the highest enzyme activity is 100%, the residual relative enzyme activity was determined to determine the optimum reaction pH.
[0037] Determination of the optimal reaction temperature: add equal amounts of free enzyme and immobilized enzyme to B-R buffer (pH3.0), and react at different temperatures (20, 30, 40, 50, 60, 70 °C), Taking the highest enzyme activity as 100%, the residual relative enzyme activity was determined to determine the optimum reaction temperature.
[0038] Determination of pH stability: equal amounts of free enzyme and immobilized enzyme were added to an equal volume of B-R buffer with pH 3.0-10.0, respectively, and incubated at 30 °C for 12 h. The untreated enzyme solution was used as a con...
Embodiment 3
[0046] Example 3 Application of immobilized laccase in degradation of soybean isoflavones
[0047] Use free enzyme and immobilized enzyme to degrade soybean isoflavones, respectively, add equal amounts of free enzyme, immobilized laccase and soybean isoflavones into the container, adjust the pH of the solution to 2.0-6.0, and then place it on a water bath shaker at 20- The reaction of 0-60min at 60°C completes the degradation step of soybean isoflavones.
[0048] Comparison of free enzyme and immobilized enzyme on the degradation of soybean isoflavones. Weigh an appropriate amount of free enzyme and immobilized enzyme, add it to the soybean isoflavone B-R buffer system with a substrate concentration of 20 mg / L, at different pH (2.0, 3.0, 4.0, 5.0, 6.0), temperature (20, 30, 40, 50, 60°C), the amount of enzyme added (1, 2, 4, 6, 8, 10, 12, 14 U / mL), and the reaction time (5, 10, 20, 30, 40 min) to degrade soybean For isoflavones, the reaction system without enzyme solution wa...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More