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Discovery and biocatalytic characterization of opine dehydrogenases by metagenome mining.
Telek, András; Molnár, Zsófia; Takács, Kristóf; Varga, Bálint; Grolmusz, Vince; Tasnádi, Gábor; Vértessy, Beáta G.
Afiliación
  • Telek A; Department of Applied Biotechnology, Budapest University of Technology and Economics, Budapest, Hungary.
  • Molnár Z; Servier Research Institute of Medicinal Chemistry, Budapest, Hungary.
  • Takács K; Institute of Molecular Life Sciences, Research Centre for Natural Sciences, HUN-REN, Budapest, Hungary.
  • Varga B; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Budapest, Hungary.
  • Grolmusz V; PIT Bioinformatics Group, Institute of Mathematics, Eötvös University, Budapest, Hungary.
  • Tasnádi G; PIT Bioinformatics Group, Institute of Mathematics, Eötvös University, Budapest, Hungary.
  • Vértessy BG; PIT Bioinformatics Group, Institute of Mathematics, Eötvös University, Budapest, Hungary.
Appl Microbiol Biotechnol ; 108(1): 101, 2024 Dec.
Article en En | MEDLINE | ID: mdl-38229296
ABSTRACT
Enzymatic processes play an increasing role in synthetic organic chemistry which requires the access to a broad and diverse set of enzymes. Metagenome mining is a valuable and efficient way to discover novel enzymes with unique properties for biotechnological applications. Here, we report the discovery and biocatalytic characterization of six novel metagenomic opine dehydrogenases from a hot spring environment (mODHs) (EC 1.5.1.X). These enzymes catalyze the asymmetric reductive amination between an amino acid and a keto acid resulting in opines which have defined biochemical roles and represent promising building blocks for pharmaceutical applications. The newly identified enzymes exhibit unique substrate specificity and higher thermostability compared to known examples. The feature that they preferably utilize negatively charged polar amino acids is so far unprecedented for opine dehydrogenases. We have identified two spatially correlated positions in their active sites that govern this substrate specificity and demonstrated a switch of substrate preference by site-directed mutagenesis. While they still suffer from a relatively narrow substrate scope, their enhanced thermostability and the orthogonality of their substrate preference make them a valuable addition to the toolbox of enzymes for reductive aminations. Importantly, enzymatic reductive aminations with highly polar amines are very rare in the literature. Thus, the preparative-scale enzymatic production, purification, and characterization of three highly functionalized chiral secondary amines lend a special significance to our work in filling this gap. KEY POINTS • Six new opine dehydrogenases have been discovered from a hot spring metagenome • The newly identified enzymes display a unique substrate scope • Substrate specificity is governed by two correlated active-site residues.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Metagenoma / Aminas Idioma: En Revista: Appl Microbiol Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Alemania
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Metagenoma / Aminas Idioma: En Revista: Appl Microbiol Biotechnol Año: 2024 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Alemania