CryoEM analysis of small plant biocatalysts at sub-2 Å resolution

Abstract

Enzyme catalysis has emerged as a key technology for developing efficient, sustainable processes in the chemical, biotechnological and pharmaceutical industries. Plants provide large and diverse pools of biosynthetic enzymes that facilitate complex reactions, such as the formation of intricate terpene carbon skeletons, with exquisite specificity. High-resolution structural analysis of these enzymes is crucial in order to understand their mechanisms and modulate their properties by targeted engineering. Although cryo-electron microscopy (cryoEM) has revolutionized structural biology, its applicability to high-resolution structural analysis of comparatively small enzymes has so far been largely unexplored. Here, it is shown that cryoEM can reveal the structures of plant borneol dehydrogenases of ∼120 kDa at or below 2 Å resolution, paving the way for the rapid development of new biocatalysts that can provide access to bioactive terpenes and terpenoids.

Accurate model and ensemble refinement using cryo-electron microscopy maps and Bayesian inference

“…Error bars show the standard deviation. b-c) Example structures and cryo-EM density maps at medium ( b : PDB 7nnh [ 94 ], 4.0 Å) and high ( c : PDB 7o6q [ 95 ], 1.88 Å) resolution. From left to right: experimental density maps and density maps predicted by the atomistic and coarse-grained forward models.…”