Efficient Transferase Engineering for SAM Analog Synthesis from Iodoalkanes

Area of Science:

• Biocatalysis
• Enzyme Engineering
• Organic Chemistry

Background:

• S-Adenosyl-l-methionine (SAM) is crucial for methyl transfer reactions.
• Expanding SAM analog chemistry requires efficient regeneration methods.

Purpose of the Study:

• Develop an enzyme engineering strategy for synthesizing diverse SAM analogs.
• Enable SAM analog regeneration using accessible iodoalkanes.

Main Methods:

• Simultaneous mutation of hydrophobic and dynamic amino acids in SAM-dependent enzymes.
• Combinatorial mutagenesis guided by natural amino acid diversity.
• High-throughput chromatographic screening for SAM analog analysis.

Main Results:

• Generated a library of highly functional enzyme mutants with significantly increased activity.
• Optimized enzymes exhibit high catalytic efficiencies (up to 31 M⁻¹s⁻¹) and stereoselectivity (>99% de).
• Enzymes successfully converted various iodoalkanes, including cyclopropyl and aromatic moieties.

Conclusions:

• The developed enzyme engineering strategy efficiently produces SAM analogs.
• This approach advances selective biocatalytic alkylation chemistry.
• Enables SAM analog regeneration with readily available reagents.