Discovery and characterization of thermophilic limonene-1,2-epoxide hydrolases from hot spring metagenomic libraries.
The epoxide hydrolases (EHs) represent an attractive option for the synthesis of chiral epoxides and 1,2-diols which are valuable building blocks for the synthesis of several pharmaceutical compounds. A metagenomic approach has been used to identify two new members of the atypical EH limonene-1,2-epoxide hydrolase (LEH) family of enzymes. These two LEHs (Tomsk-LEH and CH55-LEH) show EH activities towards different epoxide substrates, differing in most cases from those previously identified for Rhodococcus erythropolis (Re-LEH) in terms of stereoselectivity. Tomsk-LEH and CH55-LEH, both from thermophilic sources, have higher optimal temperatures and apparent melting temperatures than Re-LEH. The new LEH enzymes have been crystallized and their structures solved to high resolution in the native form and in complex with the inhibitor valpromide for Tomsk-LEH and poly(ethylene glycol) for CH55-LEH. The structural analysis has provided insights into the LEH mechanism, substrate specificity and stereoselectivity of these new LEH enzymes, which has been supported by mutagenesis studies.
['Ferrandi|Erica Elisa|EE|', 'Sayer|Christopher|C|', 'Isupov|Michail N|MN|', 'Annovazzi|Celeste|C|', 'Marchesi|Carlotta|C|', 'Iacobone|Gianluca|G|', 'Peng|Xu|X|', 'Bonch-Osmolovskaya|Elizaveta|E|', 'Wohlgemuth|Roland|R|', 'Littlechild|Jennifer A|JA|', 'Monti|Daniela|D|']
D000595:Amino Acid Sequence D001426:Bacterial Proteins D020134:Catalytic Domain D003001:Cloning, Molecular D004851:Epoxide Hydrolases D004926:Escherichia coli D006358:Hot Temperature D054892:Metagenome D008969:Molecular Sequence Data D017386:Sequence Homology, Amino Acid D014867:Water
limonene-1,2-epoxide hydrolases metagenomics stereoselectivity industrial biocatalysis protein structure