AlkB Demethylase: A Versatile Tool for RNA and DNA Demethylation

Introduction to AlkB RNA/DNA Demethylase:

AlkB, an α-ketoglutarate (α-KG) and Fe(II)-dependent dioxygenase, stands as a pivotal solution for the proficient repair of diverse alkylation damages within single-stranded or double-stranded DNA and RNA. Its capabilities extend to the removal of methylated modifications, including but not limited to m6A in RNA, and m1A, m3C, m6A, etc., in DNA. The resulting demethylated substrates are primed for downstream applications such as qPCR or high-throughput sequencing analysis. This product exemplifies a high-quality recombinant RNA/DNA demethylase, meticulously obtained through expression and purification in Escherichia coli. Notably, it is devoid of DNase and RNase contaminants, enhancing its suitability for demethylation modifications and subsequent processes like library construction and high-throughput sequencing.

Distinctive Feature:

• Impeccably free of DNase and RNase, ensuring the purity and integrity of the demethylase product.

Application Scope:

• Demethylation modification, where AlkB proves instrumental in the removal of methylated modifications.
• Subsequent library construction and high-throughput sequencing, leveraging the demethylated substrates prepared by AlkB for comprehensive genetic analyses.

Quality Assurance: 

This high-quality recombinant RNA/DNA demethylase undergoes a rigorous expression and purification process within Escherichia coli, guaranteeing its purity and effectiveness. The absence of DNase and RNase contaminants further fortifies its reliability for demanding applications in demethylation modification and advanced genetic analyses.

Utilizing AlkB in your research endeavors ensures not only precision in demethylation but also sets the stage for robust library construction and high-throughput sequencing, establishing it as an indispensable tool in molecular research.

AlkB RNA/DNA Demethylase: A Versatile Enzyme for RNA and DNA Demethylation

AlkB RNA/DNA demethylase is a bifunctional enzyme that catalyzes the removal of methyl groups from both RNA and DNA. This enzyme plays a crucial role in cellular repair mechanisms by reversing methylation-induced damage to nucleic acids. AlkB demethylase is particularly effective at removing methyl groups from 3-methyladenine (3mA) and 1-methylguanine (1meG) residues in RNA and DNA, respectively.

Mechanism of Action

The demethylation reaction catalyzed by AlkB demethylase involves a two-step process:

1. Oxidation: AlkB demethylase utilizes a conserved iron-sulfur cluster to activate molecular oxygen, forming an iron(IV)-oxo intermediate. This reactive intermediate then attacks the methyl group, converting it into an unstable aldehyde.
2. Elimination: The aldehyde moiety is subsequently eliminated through a β-elimination reaction, leaving behind an unmodified base. AlkB demethylase releases formaldehyde as a byproduct of this reaction.

Applications of AlkB Demethylase

AlkB demethylase has gained significant attention due to its wide range of potential applications:

1. Epigenetics Research: AlkB demethylase can be used to study the role of DNA methylation in gene expression and regulation. By selectively removing methyl groups from specific DNA regions, researchers can investigate the functional consequences of these modifications.
2. RNA Modification Analysis: AlkB demethylase can be employed to identify and characterize methylated RNA molecules. This enzyme can be used in conjunction with RNA sequencing techniques to generate comprehensive maps of RNA methylation patterns.
3. Drug Discovery and Development: AlkB demethylase serves as a valuable tool for screening potential epigenetic drugs. By assessing the ability of compounds to inhibit or enhance the demethylation activity of AlkB demethylase, researchers can identify promising drug candidates for various therapeutic applications.
4. Biotechnology and Synthetic Biology: AlkB demethylase can be engineered to modify methylation patterns in RNA and DNA for various biotechnology and synthetic biology applications. This enzyme can be used to generate novel RNA-based therapeutics, design synthetic gene circuits, and create organisms with altered methylation profiles.