Novel Old Yellow Enzyme Subclasses.

Area of Science:

• Biocatalysis and Enzyme Engineering
• Organic Chemistry
• Drug Discovery and Development

Background:

• Chiral drug candidate molecules necessitate efficient synthesis methods.
• Biocatalytic strategies are of high interest but limited by enzyme availability.
• Existing metal- and organocatalytic methods require complementary biocatalytic tools.

Purpose of the Study:

• To expand the enzyme toolbox by identifying novel ene reductases.
• To screen bacterial strains for double bond reduction activity.
• To characterize new ene reductases and propose novel enzyme subclasses.

Main Methods:

• Screening of 19 bacterial strains for ene reductase activity using cyclohexanone and carvone.
• Identification of 47 genes encoding putative ene reductases.
• Bioinformatic analysis and biochemical characterization of four novel ene reductases.

Main Results:

• Discovery of 47 genes coding for putative ene reductases.
• Proposal of two new Old Yellow Enzyme (OYE) subclasses: OYE class III and OYE class IV.
• Characterization revealed distinct sequence motifs and substrate scope resembling thermophilic-like OYE subclasses.

Conclusions:

• The study successfully expanded the biocatalytic toolbox with novel ene reductases.
• Two new OYE subclasses (III and IV) were identified, broadening enzyme diversity.
• These novel enzymes offer potential for efficient synthesis of chiral drug candidates.