Functional Characterization and Synthetic Application of Is2-SDR, a Novel Thermostable and Promiscuous Ketoreductase from a Hot Spring Metagenome.
biocatalysis
enzyme discovery
functional characterization
ketoreductases
metagenomics
stereoselectivity
steroids
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
12 Oct 2022
12 Oct 2022
Historique:
received:
16
09
2022
revised:
05
10
2022
accepted:
07
10
2022
entrez:
27
10
2022
pubmed:
28
10
2022
medline:
29
10
2022
Statut:
epublish
Résumé
In a metagenome mining-based search of novel thermostable hydroxysteroid dehydrogenases (HSDHs), enzymes that are able to selectively oxidize/reduce steroidal compounds, a novel short-chain dehydrogenase/reductase (SDR), named Is2-SDR, was recently discovered. This enzyme, found in an Icelandic hot spring metagenome, shared a high sequence similarity with HSDHs, but, unexpectedly, showed no activity in the oxidation of the tested steroid substrates, e.g., cholic acid. Despite that, Is2-SDR proved to be a very active and versatile ketoreductase, being able to regio- and stereoselectively reduce a diversified panel of carbonylic substrates, including bulky ketones, α- and β-ketoesters, and α-diketones of pharmaceutical relevance. Further investigations showed that Is2-SDR was indeed active in the regio- and stereoselective reduction of oxidized steroid derivatives, and this outcome was rationalized by docking analysis in the active site model. Moreover, Is2-SDR showed remarkable thermostability, with an apparent melting temperature (T
Identifiants
pubmed: 36293010
pii: ijms232012153
doi: 10.3390/ijms232012153
pmc: PMC9603792
pii:
doi:
Substances chimiques
Short Chain Dehydrogenase-Reductases
EC 1.1.1.-
Hydroxysteroid Dehydrogenases
EC 1.1.-
Ketones
0
Steroids
0
Water
059QF0KO0R
Cholic Acid
G1JO7801AE
Pharmaceutical Preparations
0
Solvents
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Biodiversity Future Center
ID : CN00000033, PNRR MUR -M4C2 -D. D. n.1034, 17.06.2022
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