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Biological macromolecules are organic compounds, predominantly composed of carbon atoms. The carbon atoms are covalently bonded with hydrogen, oxygen, nitrogen, and other minor elements. There are four major biological macromolecule classes: carbohydrates, lipids, proteins, and nucleic acids.
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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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Apollon: a deoxyribozyme that generates a yellow product.

Martin Volek1,2, Jaroslav Kurfürst1,3, Milan Kožíšek1

  • 1Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague 166 10, Czech Republic.

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Researchers developed Apollon, a novel deoxyribozyme for colorimetric assays. This self-phosphorylating enzyme converts a colorless substrate to a yellow product, enabling simple and cost-effective detection in various applications.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Colorimetric assays offer simple, inexpensive experimental monitoring.
  • Enzyme-based detection systems are crucial for biochemical analysis.

Purpose of the Study:

  • To identify and characterize a novel deoxyribozyme for colorimetric detection.
  • To develop a self-phosphorylating kinase deoxyribozyme for signal generation.

Main Methods:

  • In vitro selection and high-throughput sequencing to identify the deoxyribozyme.
  • Biochemical assays and NMR spectroscopy for characterization.
  • Engineering of deoxyribozyme variants for specific detection.

Main Results:

  • A self-phosphorylating deoxyribozyme, named Apollon, was identified.
  • Apollon converts pNPP to pNP, producing a colorimetric signal.
  • Engineered Apollon detected specific oligonucleotides and nucleases in homogenous assays.

Conclusions:

  • Apollon provides a versatile tool for colorimetric detection.
  • The deoxyribozyme enables cost-effective, high-throughput screening.
  • Apollon is suitable for applications lacking specialized equipment.