Characterization and Application of an Aspartate Dehydrogenase from Achromobacter denitrificans.
Appl Biochem Biotechnol
; 2024 Feb 22.
Article
in En
| MEDLINE
| ID: mdl-38386141
ABSTRACT
A novel gene encoding aspartate dehydrogenase (ASPDH) has been discovered in Achromobacter denitrificans. The product of this gene has a strict dependence on NADH and demonstrated significant reductive activity towards not only oxaloacetate (OAA) but also 2-ketobutyric acid. Further enzymatic characterization revealed the kinetic parameters of ASPDH for OAA and 2-ketobutyric acid were as follows Km values of 4.25 mM and 0.89 mM, Vmax values of 10.67 U mg-1 and 2.10 U mg-1, and Kcat values of 3.70 s-1 and 0.72 s-1, respectively. The enzyme also showed a dependency on metal ions, with EDTA and Cu2+ exerting strong inhibitory effects, while Ca2+ and Fe2+ exhibited pronounced enhancing effects. By utilizing a whole-cell biocatalyst system comprising glucose dehydrogenase (GDH) and ASPDH as a coupled system to replenish cofactors by oxidizing glucose, enabling the effective conversion of 2-ketobutyric acid to L-2-aminobutyric acid (L-2-ABA) with 97.2% yield.
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01-internacional
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MEDLINE
Language:
En
Journal:
Appl Biochem Biotechnol
Year:
2024
Document type:
Article
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