Weizmann Institute of Science’s team of scientists, led by Prof. To do this, they had created an arsenal of high-efficiency Kemp elimination enzymes that have amazing potential. Led by Dina Listov and Zhuofan Shen, the research team employed advanced computational methods to create enzymes that achieve catalytic rates 10 to 70 times higher than their natural counterparts.
The collaborative project further developed the unique enzyme Des27. They engineered an alternative variant Des27.7, which exhibited impressive catalytic efficiency with a k cat/K M of 12,700 M −1 s −1 and a k cat of 2.85 s −1. The researchers specifically wanted to improve the catalytic performance of enzymes and maintain their stability and efficiency.
The researchers accomplished this using a novel computational workflow to create thousands of TIM-barrel backbones via combinatorial assembly and design. From this deep library, they selectively selected 73 designs for further experimental testing. From those, 66 were successfully expressed in a soluble form. Remarkably, 14 of these designs ensured cooperative thermal denaturation behavior, reflecting their application potentials to high-stability usages.
One particularly notable variant based on Des61, for which kcat/KM was calculated, reached an impressive kcat/KM of 3,600 M −1 s −1. Additionally, it had the highest turnover rate (0.85 s−1). The team really got it to pop with a brilliant innovation by incorporating a single-point mutation in one of the designs. This simple change increased its catalytic efficiency to an impressive 123,000 M−1 s−1 and increased the kcat to 30 s −1.
This study really highlights the incredible power of computational enzyme design. Further, it showcases the exciting promise to produce enzymes with comparable efficacy and robustness to nature’s best.
“Complete computational design of high-efficiency Kemp elimination enzymes.” – Dina Listov et al
“Highly efficient enzymes designed from scratch.” – Zhuofan Shen et al