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Recombinant horseradish peroxidase: production and analytical applications
V G Grigorenko1, I P Andreeva, M Yu Rubtsova
1Lomonosov Moscow State University, Chemical Faculty, Moscow, 119991, Russia. vitaly.grigorenko@gmail.com.
Biochemistry. Biokhimiia
|April 15, 2015
Summary
Recombinant horseradish peroxidase produced in E. coli matches native enzyme performance. This genetically engineered enzyme is valuable for immunoassays like ELISA and in biosensors.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Horseradish peroxidase (HRP) is crucial for bio- and immunochemical analyses.
- Traditional HRP is sourced from plants, posing production challenges.
Purpose of the Study:
- To review the production and applications of recombinant horseradish peroxidase (rHRP).
- To highlight rHRP's utility in ELISA and amperometric sensors.
Main Methods:
- Functional expression of the HRP gene in E. coli.
- Protein refolding to achieve active recombinant enzyme.
- Genetic engineering for creating HRP conjugates.
Main Results:
- Recombinant HRP exhibits spectral and catalytic properties similar to native HRP.
- rHRP can be conjugated with protein antigens and antibody fragments.
- rHRP is effective as a marker in ELISA and amperometric sensors.
Conclusions:
- Recombinant HRP offers a viable alternative to native HRP.
- Genetic engineering enables tailored HRP conjugates for specific applications.
- rHRP is a versatile tool in biochemical and biosensor technologies.
