Method for preparing fructose lysine enzyme and application of preparing fructose lysine enzyme

Abstract

The invention provides a method for preparing fructose lysine enzyme and a glycated albumin detection kit, wherein the glycated albumin detection kit is prepared by utilizing the fructose lysine enzyme that is prepared by adopting the method. The method comprises the operation steps as follows: strains are screened so as to obtain 11 to 82 aspergillus strains of the high-activity fructose lysine enzyme; the 11 to 82 aspergillus strains are cultivated, and mycelia are collected; the mycelia are suspended in buffer solution, and then are processed through cell disruption and centrifugation, and supernatant fluid is collected; the supernatant fluid is processed through fractional precipitation by adopting ammonium sulfate solution, and deposits are collected so as to obtain crude extracts; the crude extracts are processed through hydrophobic chromatography, and eluant is collected; in addition, the collected eluant is processed through affinity chromatography, and then eluant is collected, that is, fructose lysine enzyme solution is obtained. The method has the advantages as follows: the preparation steps are simple, and the obtained fructose lysine enzyme achieves high purity, high activity and high reaction specificity.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a preparation method and application of fructose lysinase used in medical testing. Background technique [0002] Diabetes is a type of metabolic disease that occurs in people with high blood sugar, which can lead to serious damage to most systems of the body, especially nerves and blood vessels. According to the estimates of the World Health Organization, the number of people with diabetes in China ranks second in the world. Diabetes will become the most serious public health problem in China in the next 50 years. [0003] Glycated albumin (Glycated albumin, GA) refers to the product formed by the non-enzymatic glycation reaction between glucose and the N-terminal of albumin in human serum, 90% of which react with the lysine ε-NH2 residue in the albumin chain , the reaction principle is that the two first form unstable glucosylamine (Glycosylamine) or Schiff base (Schif...

AI Technical Summary

Problems solved by technology

[0013] However, the existing imported kits for the determination of glycosylated albumin on the market have the disadvantages of poor stability (in the form of dry powder), a large amount of enzyme required, and low specificity. Therefore, it is required to develop a highly active fructoaminoacidase to solve the above drawbacks.

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