Increased thermal stability and catalytic efficiency of 3-ketosteroid Δ-dehydrogenase5 from Arthrobacter simplex significantly reduces enzyme dosage in prednisone acetate biosynthesis.

The 3-ketosteroid-Δ-dehydrogenase5 (KsdD) from Arthrobacter simplex converts cortisone acetate to prednisone acetate, an important step in steroid catabolism. To achieve sustainable and efficient enzyme production, we employed computer-aided screening, structural analysis, and combinatorial experiments to identify engineered KsdD variants (M1 and M3) with dual advantages of stability and active sites. M1 had a 8.