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Literature summary for 1.4.1.1 extracted from

  • Han, S.; Shin, J.
    One-pot preparation of D-amino acids through biocatalytic deracemization using alanine dehydrogenase and omega-transaminase (2018), Catal. Lett., 148, 3678-3684 .
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
gene AlaDH, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
additional information biocatalytic deracemization of aliphatic amino acids into D-enantiomers by running cascade reactions: (1) stereoinversion of L-alanine to a D-form by L-alanine dehydrogenase and omega-transaminase and (2) regeneration of NAD+ by NADH oxidase. Under the cascade reaction conditions containing 100 mM isopropylamine and 1 mM NAD+, complete deracemization of 100 mM DL-alanine is achieved after 24 h with 95% reaction yield of D-alanine (over 99% enantiomeric excess, 52% isolation yield). AlaDH produces pyruvate from L-alanine with NAD+, NOX oxidizes NADH to NAD+, and reductive amination of the resulting pyruvate back to the amino acid in an enantiomerically opposite form by D-selective omega-transaminase (omega-TA) using isopropylamine as an amino donor cosubstrate. Method evaluation, overview Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-alanine + H2O + NAD+ Bacillus subtilis
-
pyruvate + NH3 + NADH + H+
-
?
L-alanine + H2O + NAD+ Bacillus subtilis 168
-
pyruvate + NH3 + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Bacillus subtilis Q08352
-
-
Bacillus subtilis 168 Q08352
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Bacillus subtilis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4
-
recombinant His-tagged enzyme, pH 7.0, 37°C Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-alanine + H2O + NAD+
-
Bacillus subtilis pyruvate + NH3 + NADH + H+
-
?
L-alanine + H2O + NAD+
-
Bacillus subtilis 168 pyruvate + NH3 + NADH + H+
-
?
additional information one-pot preparation of D-amino acids through biocatalytic deracemization using alanine dehydrogenase and omega-transaminase. AlaDH produces pyruvate from L-alanine with NAD+, NOX oxidizes NADH to NAD+, and reductive amination of the resulting pyruvate back to the amino acid in an enantiomerically opposite form by D-selective omega-transaminase (omega-TA) using isopropylamine as an amino donor cosubstrate Bacillus subtilis ?
-
-
additional information one-pot preparation of D-amino acids through biocatalytic deracemization using alanine dehydrogenase and omega-transaminase. AlaDH produces pyruvate from L-alanine with NAD+, NOX oxidizes NADH to NAD+, and reductive amination of the resulting pyruvate back to the amino acid in an enantiomerically opposite form by D-selective omega-transaminase (omega-TA) using isopropylamine as an amino donor cosubstrate Bacillus subtilis 168 ?
-
-

Subunits

Subunits Comment Organism
homohexamer 6 * 39700, recombinant His-tagged enzyme, SDS-PAGE Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Bacillus subtilis
NADH
-
Bacillus subtilis