Related Experiment Video
Updated: Oct 7, 2026

Structure-Guided Design and Development of Novel Cyclophilin A Inhibitors and Ganoderiol-F Derivatives: An In-Silico Approach
Published on: June 23, 2026
Engineering the α-Helix Dynamics of Hydroxysteroid Dehydrogenase by Computational Design to Balance Catalytic
Shu-Fang Li1,2,3,4, Ting Zou1,2,3,4, Sen-Yu Fu1,2,3,4
1State Key Laboratory of Green Chemical Synthesis and Conversion, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, China.
Abstract:
The stability-activity trade-off remains a major bottleneck in engineering robust biocatalysts for industrial applications. Here, we developed an integrated computational strategy to simultaneously enhance the catalytic activity and thermostability of 7α-hydroxysteroid dehydrogenase (7α-HSDH) by engineering highly flexible regions, with a particular focus on α-helices. Evolutionary conservation analysis combined with multi-temperature molecular dynamics (MD) simulations was first applied to identify weakly conserved and flexible α-helices. Subsequently, computational tools (PROSS and Pythia) were utilized to generate mutations within these flexible regions. Through modular assembly of synergistic mutations, the optimal mutant M9 (D12K/V15I/V50E/I56R/D66A/T76R/A126I/A132H/S158A/S161A/A203P/H208Q) was obtained. Compared with the WT, M9 exhibited a 10.5°C increase in melting temperature (Tm), a 2.3-fold prolongation of half-life (t1/2) at 40°C (reaching 75.3 h), a 3.14-fold improvement in catalytic efficiency (kcat/Km) to 29,032.73 mM-1 s-1, and enhanced substrate tolerance. Structural and molecular dynamics analyses indicated that the enhanced performance of M9 was associated with increased rigidity of key α-helix regions, redistribution of surface electrostatic potential, and remodeling of the substrate-binding pocket. This study thus offers a novel design framework for improving the catalytic activity, thermostability, and substrate tolerance, balancing the stability-activity trade-off.
More Related Videos
Related Concept Videos
Structure-Activity Relationships and Drug Design
SAR studies the intricate relationship between a drug's chemical structure and biological activity. It focuses on understanding how modifications to a drug's structure can influence its...
Reduction of Alkenes: Asymmetric Catalytic Hydrogenation
The metal catalyst used can be either heterogeneous or homogeneous. When hydrogenation of an alkene generates a chiral center, a pair of enantiomeric products is expected to form. However, an enantiomeric excess of one of the products can be facilitated using an enantioselective reaction or an...
Reduction of Alkenes: Catalytic Hydrogenation
Metals like palladium, platinum, and nickel are commonly used in their solid forms — fine powder on an inert surface. As these catalysts remain insoluble in the reaction mixture, they are referred to as heterogeneous catalysts.
The hydrogenation process takes place on the surface of...
Stability of Conjugated Dienes
A comparison of the enthalpies of hydrogenation of dienes reveals that conjugated dienes release less heat on hydrogenation, rendering them more stable than their nonconjugated analogs.
Oxidation of Alkenes: Syn Dihydroxylation with Osmium Tetraoxide
Introduction to Mechanisms of Enzyme Catalysis

