Method for detecting acetate content based on enzyme-linked color development method and application thereof

An enzyme-linked color development and acetate technology, which is used in measurement devices, analysis by chemical reaction of materials, and material analysis by observing the impact on chemical indicators, can solve the problem of high price of ion chromatograph and other equipment. Difficulty in large-scale promotion and use

Active Publication Date: 2021-09-07
SHANDONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the commonly used methods for the determination of acetate usually use ion chromatography for ion detection. Although it has the advantages of accurate measurement, the equipment such as ion chromatography is expensive and difficult to promote and use on a large scale.

Method used

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  • Method for detecting acetate content based on enzyme-linked color development method and application thereof
  • Method for detecting acetate content based on enzyme-linked color development method and application thereof
  • Method for detecting acetate content based on enzyme-linked color development method and application thereof

Examples

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Effect test

Embodiment 1

[0059] Preparation of succinate-CoA:acetate-CoA-transferase (succinyl-CoA:acetate CoA-transferase, aarC):

[0060] 1. Full-length cloning of aarC gene

[0061] The coding region sequence (Accession No. D13291) of the aarC gene of Acetobacter aceti was obtained from NCBI, and a pair of gene-specific primers (aarC F, aarC R) were designed. After the genomic DNA of Acetobacter is extracted, the full-length sequence of the gene is amplified by polymerase chain reaction (Polymerase Chain Reaction, PCR).

[0062] The PCR reaction system was as follows: primers (10 μmol L -1 ) each was 3 μL, 2x Besttaq Master Mix (Applied Biological Materials Inc.) was 25 μL, cDNA was 0.3 μL, ddH 2 O is 18.7 μL. The PCR program is: 94°C for 3min; 94°C for 10s, 56°C for 45s, 72°C for 1min, 10 cycles; 94°C for 10s, 53°C for 45s, 72°C for 1min, 10 cycles; 94°C for 10s, 49°C for 45s, 72°C 1min, 15 cycles; 10min at 72°C. The PCR products were detected by 1% agarose gel electrophoresis to determine wh...

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Abstract

The invention provides a method for detecting acetate content based on an enzyme-linked color development method and application thereof, and belongs to the technical field of compound detection. Researches find that succinyl coenzyme A: acetic acid CoA transferase can take succinyl coenzyme A and acetate as substrates to form acetyl coenzyme A and succinic acid, and subsequently, acetyl coenzyme A is oxidized under the action of acyl coenzyme A oxidase so as to generate enoyl coenzyme A and H2O2. According to the invention, the acetic acid content is indirectly determined by using a hydrogen peroxide (H2O2) content detection kit to determine H2O2, and the method has advantages of high detection sensitivity, low detection cost, and good practical application value.

Description

technical field [0001] The invention belongs to the technical field of compound detection, and in particular relates to a method for detecting acetate content based on an enzyme-linked chromogenic method and an application thereof. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Acetic acid is a bulk chemical product and one of the important organic acids. It is mainly used in the production of vinyl acetate, acetic anhydride, acetate and cellulose acetate. It is one of the important raw materials for pesticides, medicines, textile printing and dyeing, and synthetic fibers. For example, polyvinyl acetate can be used to prepare films and adhesives, and is also a raw materi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
CPCG01N21/78G01N2021/775
Inventor 孔令明张雷陈知雨徐瑜聪
Owner SHANDONG UNIV
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