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A method for preparing a glutamate sensing electrode with a wide detection linear range

A linear range, sensing electrode technology, applied in measurement devices, material electrochemical variables, material analysis by electromagnetic means, etc., can solve the problems of narrow detection range, long detection time and low detection efficiency of glutamate biosensors , to achieve the effect of wide linear detection range, excellent storage stability and high test accuracy

Active Publication Date: 2022-03-08
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, glutamic acid biosensors have problems such as long detection time, narrow detection range, low detection efficiency and weak anti-interference ability.
The content of glutamic acid in the glutamic acid fermentation liquid in industrial production is relatively high, and the existing glutamic acid biosensors on the market still cannot realize real-time detection of it, and cannot be used in actual production

Method used

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  • A method for preparing a glutamate sensing electrode with a wide detection linear range
  • A method for preparing a glutamate sensing electrode with a wide detection linear range
  • A method for preparing a glutamate sensing electrode with a wide detection linear range

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preparation example Construction

[0055] In one embodiment, the preparation method of glutamate oxidase in the step (7) comprises the following steps:

[0056] (1) The glutamic acid oxidase gene sequence is synthesized and constructed on a plasmid to obtain a recombinant plasmid;

[0057] (2) Mix the plasmid with the competent large intestine, put it on ice for 30 minutes, heat shock at 42°C for 90 seconds, put it on ice again for 2 minutes, then add it to LB liquid medium for culture, and then take an appropriate amount of cultured bacteria liquid to spread Cultivate overnight on LB plates to obtain glutamic acid oxidase-producing bacteria;

[0058] (3) Inoculate the glutamic acid oxidase-producing bacteria in LB liquid medium and cultivate overnight; transfer the culture solution into LB liquid medium for fermentation and culture, then add lactose or isopropyl-β-D-thiopyril Galactopyranoside induced expression, and the cells were collected by centrifugation;

[0059] (4) Wash the bacterial cells obtained i...

Embodiment 1

[0075] Embodiment 1 of the present invention provides a method for preparing a glutamate sensing electrode with a wide detection linear range, and the method for preparing a glutamate sensing electrode with a wide detection linear range includes the following steps:

[0076] (1) 1.8mmol FeCl 3 , 1.8 mmol K 3 [Fe(CN) 6 ] was dissolved in 50mL of a solution containing 0.08mol HCl and 0.08mol KCl;

[0077] (2) The solution obtained in step (1) is passed through argon for 8 minutes;

[0078] (3) The bare glassy carbon electrode with a diameter of 2 mm is sequentially polished and polished by alumina powder with a diameter of 0.3 μm and 0.05 μm to obtain a glassy carbon electrode with a mirror surface;

[0079] (4) Under the condition of +400mV, place the electrode obtained in step (3) in the solution obtained in step (2) for 35s;

[0080] (5) Using a solution containing 0.08mol / L HCl and 0.08mol / L KCl as the electrolyte, cycle the electrode obtained in step (4) for 18 times at...

Embodiment 2

[0090] Embodiment 2 of the present invention provides a method for preparing a glutamate sensing electrode with a wide detection linear range, and the method for preparing a glutamate sensing electrode with a wide detection linear range includes the following steps:

[0091] (1) 2.3mmol FeCl 3 , 2.3 mmol K 3 [Fe(CN)6 ] was dissolved in 50 mL of a solution containing 0.12 mol HCl and 0.12 mol KCl;

[0092] (2) The solution obtained in step (1) is passed through argon for 12 minutes;

[0093] (3) The bare glassy carbon electrode with a diameter of 2 mm is polished and polished by alumina powder with a diameter of 0.3 μm and 0.05 μm in sequence to obtain a glassy carbon electrode with a mirror surface;

[0094] (4) Under the condition of +400mV, place the electrode obtained in step (3) in the solution obtained in step (2) for 45s;

[0095] (5) Using a solution containing 0.12mol / L HCl and 0.12mol / L KCl as the electrolyte, cycle the electrode obtained in step (4) for 23 times a...

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Abstract

The present invention relates to the field of preparation of glutamate sensing electrodes, more specifically, the present invention relates to a method for preparing a glutamate sensing electrode with a wide detection linear range, and the preparation of a glutamate sensing electrode with a wide detection linear range The method comprises: (1) FeCl 3 、K 3 [Fe(CN) 6 ] was dissolved in 50mL solution containing HCl and KCl; (2) the solution obtained in step (1) was passed through argon for 8-12min; (3) the bare glassy carbon electrode with a diameter of 2mm was polished by alumina powder to obtain a mirror surface (4) Under the condition of +400mV, the electrode obtained in step (3) is placed in the solution obtained in step (2); (5) with the solution containing HCl and KCl as the electrolyte, the step (4) ) to activate the electrode obtained; (6) drying the electrode obtained in step (5) at 100-110° C. for 55-65 min and cooling to room temperature; (7) mixing glutamic acid oxidase with glutaraldehyde solution to obtain a mixed solution , the mixed solution is titrated on the electrode obtained in step (6).

Description

technical field [0001] The invention relates to the field of preparation of glutamate sensing electrodes, and more specifically, the invention relates to a method for preparing glutamate sensing electrodes with a wide detection linear range. Background technique [0002] As an important industrial raw material, glutamic acid is widely used in the fields of daily chemical, medicine and food. As a natural plant ingredient, glutamic acid can be used in the production of daily cosmetics such as hair care, skin care cosmetics, etc. Glutamic acid has a good effect on the prevention and treatment of hepatic coma, liver protection and treatment of concussion or nerve damage. The main ingredient of common food fresheners in daily life is glutamic acid. Glutamic acid can also be used to synthesize polyglutamic acid as a raw material for the production of environmentally friendly plastics. At present, the main production method of glutamic acid is microbial fermentation, with an ann...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/30G01N27/327
CPCG01N27/308G01N27/3275
Inventor 周杰庄妮莎吕紫瑶董维亮薛瑞姜岷
Owner NANJING TECH UNIV