| dc.creator |
Ferrandi, EE |
|
| dc.creator |
Sayer, C |
|
| dc.creator |
De Rose, SA |
|
| dc.creator |
Guazzelli, E |
|
| dc.creator |
Marchesi, C |
|
| dc.creator |
Saneei, V |
|
| dc.creator |
Isupov, MN |
|
| dc.creator |
Littlechild, JA |
|
| dc.creator |
Monti, D |
|
| dc.date |
2018-11-30T11:21:01Z |
|
| dc.date |
2018-10-16 |
|
| dc.date |
2018-11-30T11:21:01Z |
|
| dc.date.accessioned |
2022-05-27T01:03:18Z |
|
| dc.date.available |
2022-05-27T01:03:18Z |
|
| dc.identifier |
Vol. 6, article 144 |
|
| dc.identifier |
10.3389/fbioe.2018.00144 |
|
| dc.identifier |
http://hdl.handle.net/10871/34937 |
|
| dc.identifier |
2296-4185 |
|
| dc.identifier |
Frontiers in Bioengineering and Biotechnology |
|
| dc.identifier.uri |
http://localhost:8080/xmlui/handle/CUHPOERS/241938 |
|
| dc.description |
This is the final version. Available from Frontiers Media via the DOI in this record. |
|
| dc.description |
Two novel epoxide hydrolases (EHs), Sibe-EH and CH65-EH, were identified in the metagenomes of samples collected in hot springs in Russia and China, respectively. The two α/β hydrolase superfamily fold enzymes were cloned, over-expressed in Escherichia coli, purified and characterized. The new EHs were active toward a broad range of substrates, and in particular, Sibe-EH was excellent in the desymmetrization of cis-2,3-epoxybutane producing the (2R,3R)-diol product with ee exceeding 99%. Interestingly these enzymes also hydrolyse (4R)-limonene-1,2-epoxide with Sibe-EH being specific for the trans isomer. The Sibe-EH is a monomer in solution whereas the CH65-EH is a dimer. Both enzymes showed high melting temperatures with the CH65-EH being the highest at 85°C retaining 80% of its initial activity after 3 h thermal treatment at 70°C making it the most thermal tolerant wild type epoxide hydrolase described. The Sibe-EH and CH65-EH have been crystallized and their structures determined to high resolution, 1.6 and 1.4 Å, respectively. The CH65-EH enzyme forms a dimer via its cap domains with different relative orientation of the monomers compared to previously described EHs. The entrance to the active site cavity is located in a different position in CH65-EH and Sibe-EH in relation to other known bacterial and mammalian EHs. |
|
| dc.language |
en |
|
| dc.publisher |
Frontiers Media |
|
| dc.relation |
https://www.ncbi.nlm.nih.gov/pubmed/30386778 |
|
| dc.rights |
© 2018 Ferrandi, Sayer, De Rose, Guazzelli, Marchesi, Saneei, Isupov, Littlechild and Monti. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY): https://creativecommons.org/licenses/by/4.0/. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
|
| dc.rights |
https://creativecommons.org/licenses/by/4.0/ |
|
| dc.subject |
epoxide hydrolase |
|
| dc.subject |
industrial biocatalysis |
|
| dc.subject |
metagenomics |
|
| dc.subject |
protein structure |
|
| dc.subject |
stereoselectivity |
|
| dc.title |
New Thermophilic α/β Class Epoxide Hydrolases Found in Metagenomes From Hot Environments |
|
| dc.type |
Article |
|