Enhanced D-Lactate Dehydrogenase Activity Achieved Through Evolutionary Engineering: A Comprehensive Analysis of 45 Mutants

This research employed the CSFp strategy to evolve D-lactate dehydrogenase (D-Lac) activity, resulting in the generation of 45 distinct mutants. A noteworthy 69% of these mutants exhibited enhanced D-Lac activity, with 16 surpassing the baseline activity by an order of magnitude. Among these high-performing variants, mutant N emerged as the most successful, demonstrating a remarkable 56-fold increase in D-Lac activity. This comprehensive analysis underscores the effectiveness of the CSFp approach in generating a diverse pool of mutants with significantly improved D-Lac activity, highlighting its potential for biocatalytic applications.