Recombinant glycerol kinase: solving the problem of detection interference and improving the accuracy of results

In clinical diagnosis and other fields, glycerol kinase is a key tool enzyme commonly used for glycerol concentration detection. However, glycerol kinases derived from traditional natural bacterial strains are subject to long-term interference by enzymes, leading to detection errors. Driven by genetic engineering technology, recombinant glycerol kinase provides a new solution for precise detection by reducing the content of interfering enzymes.

The 'congenital defect' of natural glycerol kinase: interference with enzymes causing detection bias

Natural glycerol kinases are mostly extracted from bacteria and yeast, which contain a large number of metabolic enzymes, with NADH oxidase and catalase being the main disruptors. In the detection of glycerol kinase, the signal is generated by the oxidation-reduction reaction of coenzyme NADH. The mixed NADH oxidase will decompose NADH, resulting in low detection results; If the reaction involves hydrogen peroxide, catalase will decompose it, disrupting signal stability and causing significant data fluctuations.

Glycerokinase

More importantly, the purification of natural glycerol kinase is difficult. Due to the small difference in molecular weight and charge between it and interfering enzymes, traditional purification methods cannot completely separate them. Even after multiple rounds of purification, residual interfering enzymes may still exceed the standard, posing a hidden danger to detection.

Precision improvement of recombinant technology: reducing interfering enzymes from the source

The breakthrough of recombinant glycerol kinase stems from the precise control of enzyme preparation by genetic engineering, with the core being to enable engineered bacteria to "specifically" express the target enzyme and cut off the pathway of interfering enzyme production.

Researchers have cloned genes and introduced only the gene encoding glycerol kinase into engineering bacteria such as Escherichia coli and Pichia pastoris. The modified engineering bacterial genome only contains glycerol kinase expression sequences and does not carry interfering enzyme coding genes, fundamentally avoiding interference with enzyme synthesis.

Meanwhile, the expression and purification of recombinant glycerol kinase are more controllable. Engineering bacteria efficiently express glycerol kinase in optimized environments, and the expression level is much higher than that of natural strains. During purification, high-purity separation can be achieved by combining affinity chromatography and other techniques due to the low amount of impurity proteins. The data shows that the purity of high-quality recombinant glycerol kinase exceeds 95%, and the residual interference enzyme is reduced below the detection limit, solving the problem of natural enzyme purification.

The application value of recombinant enzymes: empowering precise detection in multiple fields

Recombinant glycerol kinase has demonstrated value in multiple fields due to its low interference and high stability advantages. In clinical practice, the reagent kit made with it can reduce the error rate of serum glycerol detection and provide reliable data for early diagnosis of metabolic diseases; In addition, the batch stability of recombinant glycerol kinase is high. Natural enzymes are greatly affected by cultivation and processing, resulting in significant batch differences; The recombinant enzyme undergoes a standardized process with consistent performance parameters, reducing the difficulty of quality control for testing institutions.

From the interference problem of natural enzymes to the precise breakthrough of recombinant enzymes, the upgrade of glycerol kinase technology not only reflects the innovation of genetic engineering in enzyme engineering, but also promotes the development of detection technology towards high precision and stability. In the future, with the optimization of recombinant technology, recombinant glycerol kinase is expected to make efforts in more fields, injecting vitality into the precision detection industry.

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Hubei Xindesheng Material Technology Co., Ltd. has always adhered to the production and operation philosophy of "quality first, technology leadership" and "innovation and pursuit of excellence", continuously researching and developing reagents needed in the diagnostic field. Since its establishment in 2005, it has a history of nearly 20 years and is deeply trusted and supported by users. The diagnostic enzyme reagents independently developed and produced by Desheng have been sold to various parts of the world. If you have any related procurement needs in the near future, please feel free to contact us!