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Detection apparatus for rapid calculation of generation amount of gamma-aminobutyric acid in biotransformation

A technology of aminobutyric acid and a detection device, which is applied in the field of chemical instruments, can solve problems such as high cost, complicated operation, and small scale, and achieve the effects of low cost, simple operation, and short time consumption

Inactive Publication Date: 2013-08-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The technical problem to be solved in the present invention is: aiming at the shortcomings of the current GABA determination method, such as time-consuming, high cost, complicated operation, and small scale, a detection device specially used for determining the content of GABA produced in biotransformation has been invented.

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  • Detection apparatus for rapid calculation of generation amount of gamma-aminobutyric acid in biotransformation

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Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1: operate according to the following steps: a. set the parameters of the controller: pH4.6, add 0.1M hydrochloric acid solution; b. add 20mmol of glutamic acid to the reactor, add deionized water to a reaction volume of 100mL; c . Adjust to pH 4.6; d. Add 10mL glutamic acid decarboxylase solution (enzyme activity 100U) into the reactor to start the reaction. After 10 minutes, record the accumulative volume of added hydrochloric acid as 79.5mL, use the formula W=(n×V× k ×103.1) The amount of GABA produced was calculated to be 409.8 mg, and the detection result of GABA by HPLC was 417 mg, with an error rate of 1.8%.

Embodiment 2

[0023] Embodiment 2: operate according to the following steps: a. set the parameters of the controller: pH4.8, add 0.5M hydrochloric acid solution; b. add 5 mmol of glutamic acid to the reactor, add deionized water to 200mL; c. adjust To pH4.8; d. Add 20g of wet cells (enzyme activity 500U) into the reactor to start the reaction. After 10 minutes, record the accumulative volume of added hydrochloric acid as 57.3mL, use the formula W=(n×V× k ×103.1) The amount of GABA produced was calculated to be 1477mg, and the result of GABA detection by HPLC was 1489.4mg, with an error rate of 0.9%.

Embodiment 3

[0024] Embodiment 3: operate according to the following steps: a. set the parameters of the controller: pH4.7, add 0.25M hydrochloric acid solution; b. add 100mmol of glutamic acid to the reactor, add deionized water to 500mL; c. adjust To pH4.7; d. Add 80g of wet cells (enzyme activity 2000U) into the reactor to start the reaction. After 10 minutes, record the accumulative volume of added hydrochloric acid as 331.3mL, use the formula W=(n×V× k ×103.1) The amount of GABA produced was calculated to be 4.2696g, and the detection result of GABA by HPLC method was 4.32g, with an error rate of 1.1%.

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Abstract

The invention provides a detection apparatus for rapid calculation of generation amount of gamma-aminobutyric acid (GABA) in biotransformation, which belongs to the technical field of chemical apparatuses. The main body of the apparatus comprises 1) a pH induction device, 2) a digital transformation device, 3) a liquid dropping flask, 4) a controller and 5) a reactor. The digital transformation device can record the volume of an added acid, the controller can set two parameters, i.e., the pH value of a reaction and the concentration of the added acid, and the generation amount of GABA can be calculated by a program based on the volume of the added acid and is directly displayed on the controller. Protons are irreversibly consumed in a glutamic acid decarboxylase catalytic reaction; and itis proportional relationship between the number of consumed protons and GABA that the apparatus is used. The apparatus provided in the invention overcomes the defects of a long detection period, highcost and complicated operation in conventional detection of GABA in biotransformation. When the apparatus provided in the invention is used for detection of GABA in biotransformation, the advantages of no need for treatment of samples, simple and practicable operation, a short detection period and low cost are obtained.

Description

technical field [0001] The invention discloses a detection device for rapidly calculating the production amount of gamma-aminobutyric acid in a biotransformation reaction, which belongs to the technical field of chemical instruments. Background technique [0002] Gamma-aminobutyric acid (GABA, Gamma-aminobutyric acid) is a naturally occurring non-protein amino acid. In mammals, GABA is an inhibitory neurotransmitter that mediates more than 40% of inhibitory nerve signals. GABA has the functions of improving brain vitality, lowering blood pressure, stabilizing the spirit, improving liver and kidney functions, etc., so it has broad application prospects in functional foods. [0003] Glutamate decarboxylase (GAD, Glutamate decarboxylase) is a kind of pyridoxal phosphate (PLP) enzyme, which is an important biological enzyme in the process of GABA biosynthesis. GAD widely exists in microorganisms, plant tissues and advanced mammals, but its existence in cells is very small. [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/36C12M1/34C12Q1/527
CPCC12M41/32C12M41/26
Inventor 江波缪铭张天萌张涛沐万孟
Owner JIANGNAN UNIV