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Applied Microbiology and Biotechnology

, Volume 21, Issue 1–2, pp 7–15 | Cite as

d-2-hydroxyisocaproate dehydrogenase from Lactobacillus casei

A new enzyme suitable for stereospecific reduction of 2-ketocarboxylic acids
  • Werner Hummel
  • Horst Schütte
  • Maria-Regina Kula
Biotechnology

Summary

The new enzyme d-2-hydroxyisocaproate dehydrogenase (NAD+-dependent) was detected in strains of the genus Lactobacillus and related genera. Straight and branched chain aliphatic as well as aromatic 2-ketocarboxylic acids are stereospecifically reduced to the corresponding d-2-hydroxycarboxylic acids according to the following equation:R-CO-COOH + NADH + H+ ⇌ R-CHOH-COOH + NAD+

The enzyme is called d-hydroxyisocaproate dehydrogenase by us because 2-ketoisocaproate is the substrate with the lowest KM-value. NAD(H) as a cofactor cannot be replaced by NADP(H). Because of its broad substrate specificity we chose the strain Lactobacillus casei ssp. pseudoplantarum (DSM 20 008) for enzyme production and characterization. d-2-hydroxyisocaproate dehydrogenase could be purified 180-fold starting with 500 g of wet cells.

The purification procedure involved liquid-liquid extraction with aqueous two-phase systems and ion-exchange chromatography. At this stage the enzyme has a specific activity of 25 U/mg and can be used for technical applications. Further purification up to a homogeneous protein with a specific activity of 110 U/mg can be achieved by chromatography on Amberlite CG 50 at pH 3.5. Properties important for technical application of the d-HicDH were investigated, especially the substrate specificity and the optimum pH- and temperature ranges for activity and stability of the catalist.

Keywords

Enzyme NADH Lactobacillus Substrate Specificity Enzyme Production 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Werner Hummel
    • 1
  • Horst Schütte
    • 1
  • Maria-Regina Kula
    • 1
  1. 1.Gesellschaft für Biotechnologische Forschung mbHBraunschweigFederal Republic of Germany

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