Method for regenerating coenzyme-reduced glutathione based on electrochemistry and enzyme electrode

An enzyme electrode and electrode technology, applied in the field of enzyme electrodes, can solve the problems of low regeneration efficiency

Inactive Publication Date: 2019-04-19
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The photochemical regeneration method has not yet achieved ideal results, and its regeneration efficiency is very low, but it has broad potential application value

Method used

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  • Method for regenerating coenzyme-reduced glutathione based on electrochemistry and enzyme electrode
  • Method for regenerating coenzyme-reduced glutathione based on electrochemistry and enzyme electrode
  • Method for regenerating coenzyme-reduced glutathione based on electrochemistry and enzyme electrode

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, for the preparation method of the enzyme electrode that is used to process DCM-containing wastewater, the following steps are carried out successively:

[0033] 1) Using graphite as the conductive material, the graphite is polished on the suede with alumina powders with diameters of 1.0, 0.3, 0.05 and 0.01 μm in sequence (smooth polishing is enough), and then ultrasonically cleaned in ultrapure water for 5 minutes to obtain A graphite substrate with a clean surface (about 1.5 cm in diameter); take it out and dry it naturally until there is no visible moisture on the surface to obtain an electrode substrate.

[0034] 2), using recombinant engineering bacteria to prepare DCM dehalogenase pure enzyme solution:

[0035] ① First, the recombinant engineering bacteria were prepared by using the DCM-degrading bacteria Methylobacterium H13 (CCTCC No: M 2010121), as follows:

[0036] The DCM dehalogenase gene was obtained by PCR cloning from the whole genome of the...

Embodiment 2

[0062] Embodiment 2, a kind of enzyme electric coupling dechlorination system device for processing DCM, such as figure 1 As shown, it consists of three parts: temperature control system, electrolytic reactor and pH adjustment system.

[0063] The electrolytic reactor includes a power source 1 and a reaction tank 15 , the power source 1 provides current for the electro-biological enzyme system, and the current is monitored in real time by the ammeter 2 . The reaction tank 15 is placed on the constant temperature magnetic stirrer 8, and the anion exchange membrane 7 is provided in the reaction tank 15, thereby the reaction tank 15 is separated to form a cathode electrolyzer and an anode electrolyzer, and a constant temperature magnetic force is provided in the cathode electrolyzer. The magnetic rotor 10 connected to the stirrer 8, under the drive of the constant temperature magnetic stirrer 8, the magnetic rotor 10 uniformly stirs the liquid in the reaction tank 15; Graphite e...

Embodiment 3

[0067] Embodiment 3, utilize the system device as described in embodiment 2, inject the 0.01M phosphate buffer solution of 100mL in the anode electrolyzer, inject the 0.01M phosphate buffer solution of 200mL in the cathode electrolyzer and add 3mg as coenzyme Reduced glutathione (GSH), 1.5 μ L DCM, adjust the pH to be 7 (regulate with a dilute sulfuric acid solution with a concentration of 20%), adjust the temperature of the water bath to be 35 ° C, and use a power supply control circuit with a current of 15 mA for constant current electrolysis 2h.

[0068] At this time, the dosage of DCM dechlorination enzyme is 28 mg; in the cathodic electrolyzer, the ratio of GSH to waste water containing DCM is 3 mg / 200 mL=15 mg / L.

[0069] Such as figure 2 As shown, the liquid in the cathode electrolyzer is detected, and the DCM concentration and degradation rate at different times are as follows in Table 1:

[0070] Table 1

[0071] Degradation time (min)

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Abstract

The invention discloses an enzyme electrode, which is prepared by the following steps of: preparing recombinant engineering bacteria by using a DCM degrading bacterium Methylobacterium H13, and obtaining DCM dehalogenase pure enzyme solution through induction culture expression and purification; mixing the DCM dehalogenase pure enzyme solution with sodium alginate to obtain an embedding material;spreading the embedding material on the surface of an electrode matrix, and fixing the embedding material by Ca (NO3)2 and the like to obtain the enzyme electrode. The invention also provides a methodfor regenerating coenzyme-reduced glutathione based on electrochemistry by using the enzyme electrode, which comprises the following steps of: injecting DCM-containing wastewater into a cathode electrolytic tank, and performing constant current electrolysis on the reduced glutathione GSH serving as coenzyme. The method utilizes electrochemical technology and a medium of the enzyme electrode to provide sufficient electrons for regenerating coenzyme GSH and promote GSSG to regenerate GSH.

Description

technical field [0001] The invention relates to an electrochemical-based coenzyme-reduced glutathione regeneration method and an enzyme electrode. Background technique [0002] Glutathione widely exists in animals, plants and microorganisms, and is the most abundant small molecule thiol compound in cells. Reduced glutathione (GSH) is the main component of non-protein sulfhydryl groups in cells, which is formed by condensation of glutamic acid (Glu), cysteine ​​(Cys) and glycine (Gly) through peptide bonds Tripeptide has a relative molecular mass of 307.32 and an isoelectric point of 5.93. It is a white crystal at room temperature and is easily soluble in water, low-concentration ethanol aqueous solution, liquid ammonia and dimethylformamide. It exists in large quantities in living organisms and plays a major role. [0003] The existing regeneration methods include enzymatic method, electrochemical method to regenerate coenzyme, photochemical method and so on. The advantage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10C12N11/04C12N15/70C12N9/14C12P21/02
CPCC12N9/14C12N11/04C12N11/10C12N11/14C12N15/70C12P21/02C12Y308/01005C07K5/0215
Inventor 於建明汤育炜吴朦王佳哲宋永泉胡俊成卓韦
Owner ZHEJIANG UNIV OF TECH
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