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Reagent system and method for testing activity of phenylalnine ammonialyase

A technology for phenylalanine ammonia lyase and activity determination, which is applied in biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problems affecting the accuracy of determination results, interference, etc., and achieves extended and accurate fluorescence retention time. Active and sensitive effects

Inactive Publication Date: 2015-04-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this detection method is simple, many substances can seriously interfere with the UV spectrophotometry and affect the accuracy of the measurement results.

Method used

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  • Reagent system and method for testing activity of phenylalnine ammonialyase
  • Reagent system and method for testing activity of phenylalnine ammonialyase
  • Reagent system and method for testing activity of phenylalnine ammonialyase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1 is the mensuration of jujube phenylalanine ammonia-lyase activity, and the molar concentration of assay reagent composition is as follows:

[0075] Buffer: 100mmol / L, pH 8.8 tris-hydrochloric acid solution;

[0076] Reagent 1: L-phenylalanine 30mmol / L, prepared with the above buffer as a solvent;

[0077] Reagent 2: succinic acid 150mmol / L, prepared with the above buffer as a solvent;

[0078] Reagent 3: Glycyl-L-leucine 5mmol / L, prepared with the above buffer as a solvent;

[0079] Reagent 4: ninhydrin 50mmol / L, prepared with the above buffer as a solvent;

[0080] Stop solution 1: hydrochloric acid 6.0mol / L, prepared with the above buffer as a solvent;

[0081] Stop solution 2: Potassium sodium tartrate 10mmol / L, sodium carbonate 200mmol / L, copper sulfate 20mmol / L, prepared with the above buffer as a solvent;

[0082] Calibration solution: 20mmol / L L-phenylalanine standard solution, prepared with the above buffer as a solvent.

[0083] Embodiment 1 a...

Embodiment 2

[0102] Embodiment 2 is the mensuration of Malin jujube phenylalanine ammonia solution activity, and the molar concentration of assay reagent composition is as follows:

[0103] Buffer: 10mmol / L, pH 9.0 tris-hydrochloric acid solution;

[0104] Reagent 1: L-phenylalanine 10mmol / L, prepared with the above buffer as a solvent;

[0105] Reagent 2: succinic acid 200mmol / L, prepared with the above buffer as a solvent;

[0106]Reagent 3: Glycyl-L-leucine 50mmol / L, prepared with the above buffer as a solvent;

[0107] Reagent 4: ninhydrin 100mmol / L, prepared with the above buffer as a solvent;

[0108] Stop solution 1: hydrochloric acid 6.0mol / L, prepared with the above buffer as a solvent;

[0109] Stop solution 2: Potassium sodium tartrate 100mmol / L, sodium carbonate 500mmol / L, copper sulfate 100mmol / L, prepared with the above buffer as a solvent;

[0110] Calibration solution: 20mmol / L L-phenylalanine standard solution, prepared with the above buffer as a solvent.

[0111] Emb...

Embodiment 3

[0130] Embodiment 3 is the mensuration of plum fruit phenylalanine ammonia-lyase activity, and the molar concentration of assay reagent composition is as follows:

[0131] Buffer: 200mmol / L, pH 8.0 tris-hydrochloric acid solution;

[0132] Reagent 1: L-phenylalanine 200mmol / L, prepared with the above buffer as a solvent;

[0133] Reagent 2: succinic acid 200mmol / L, prepared with the above buffer as a solvent;

[0134] Reagent 3: Glycyl-L-leucine 10mmol / L, prepared with the above buffer as a solvent;

[0135] Reagent 4: ninhydrin 30mmol / L, prepared with the above buffer as a solvent;

[0136] Stop solution 1: hydrochloric acid 6.0mol / L, prepared with the above buffer as a solvent;

[0137] Stop solution 2 (basic copper solution): Potassium sodium tartrate 1.0mmol / L, sodium carbonate 100mmol / L, copper sulfate 1.0mmol / L, prepared with the above buffer as a solvent;

[0138] Calibration solution: 20mmol / L L-phenylalanine standard solution, prepared with the above buffer as a s...

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Abstract

The invention belongs to the field of enzymatic activity analysis and particularly relates to a reagent system and method for testing the activity of phenylalnine ammonialyase. The reagent system comprises the following reagents: a buffer solution, a reagent 1, a reagent 2, a reagent 3, a reagent 4, a stop solution 1, a stop solution 2 and a calibrating solution. The method comprises the following steps: enzyme solution extraction, enzymatic reaction, enzyme activity assay and phenylalnine ammonialyase activity calculation. Compared with the conventional method, the method provided by the invention can effectively eliminate interference, so that a testing result can more accurately reflect the activity of phenylalnine ammonialyase.

Description

technical field [0001] The invention belongs to the field of enzymatic activity analysis, in particular to a reagent system and a method for measuring the activity of plant phenylalanine ammonia-lyase. Background technique [0002] Phenylalanine ammonia-lyase (L-phenylalanine ammonia-lyase, PAL) widely exists in various plants, and is the key enzyme and rate-limiting enzyme in the metabolism of phenylpropanoids in plants. It also plays an important role in the control of browning of plant tissues such as fruits and vegetables, disease control, and storage and preservation. Phenylalanine ammonia lyase is also widely used in the fields of food bioengineering and medicine. [0003] At present, the detection method of plant phenylalanine ammonia-lyase activity is mainly ultraviolet spectrophotometer assay, that is, L-phenylalanine is used as a substrate, and the product trans-cinnamon is generated under the action of phenylalanine ammonia-lyase. acid, the product has an absorp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/527
Inventor 曹建康丁薪源王睿袁树枝王姣谢芳潘寒姁姜微波
Owner CHINA AGRI UNIV
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