4IO0

Crystal structure of F128A mutant of an epoxide hydrolase from Bacillus megaterium complexed with its product (R)-3-[1]naphthyloxy-propane-1,2-diol

Summary for 4IO0

• Entry DOI: 10.2210/pdb4io0/pdb
• Related: 4G00 4G02 4INZ
• Descriptor: Soluble epoxide hydrolase, SULFATE ION, (2R)-3-(naphthalen-1-yloxy)propane-1,2-diol, ... (4 entities in total)
• Functional Keywords: a/b hydrolase fold, epoxide hydrolase, hydrolase
• Biological source: Bacillus megaterium
• Total number of polymer chains: 2
• Total formula weight: 71049.77

Authors

Kong, X.D.,Zhou, J.H.,Xu, J.H. (deposition date: 2013-01-07, release date: 2014-02-12, Last modification date: 2023-11-08)

Primary citation

Kong, X.D.,Yuan, S.,Li, L.,Chen, S.,Xu, J.H.,Zhou, J.H.
Engineering of an epoxide hydrolase for efficient bioresolution of bulky pharmaco substrates.
Proc.Natl.Acad.Sci.USA, 111:15717-15722, 2014

PubMed Abstract: Optically pure epoxides are essential chiral precursors for the production of (S)-propranolol, (S)-alprenolol, and other β-adrenergic receptor blocking drugs. Although the enzymatic production of these bulky epoxides has proven difficult, here we report a method to effectively improve the activity of BmEH, an epoxide hydrolase from Bacillus megaterium ECU1001 toward α-naphthyl glycidyl ether, the precursor of (S)-propranolol, by eliminating the steric hindrance near the potential product-release site. Using X-ray crystallography, mass spectrum, and molecular dynamics calculations, we have identified an active tunnel for substrate access and product release of this enzyme. The crystal structures revealed that there is an independent product-release site in BmEH that was not included in other reported epoxide hydrolase structures. By alanine scanning, two mutants, F128A and M145A, targeted to expand the potential product-release site displayed 42 and 25 times higher activities toward α-naphthyl glycidyl ether than the wild-type enzyme, respectively. These results show great promise for structure-based rational design in improving the catalytic efficiency of industrial enzymes for bulky substrates.

PubMed: 25331869
DOI: 10.1073/pnas.1404915111

Experimental method

X-RAY DIFFRACTION (2.9 Å)