Biotechnology Letters

, Volume 40, Issue 7, pp 1135–1147 | Cite as

Discovery of an acidic, thermostable and highly NADP+ dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929

  • Saadet Alpdağtaş
  • Sevil Yücel
  • Handan Açelya Kapkaç
  • Siqing Liu
  • Barış Binay
Original Research Paper



To identify a robust NADP+ dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929 (LbFDH) with unique biochemical properties.


A new NADP+ dependent formate dehydrogenase gene (fdh) was cloned from genomic DNA of L. buchneri NRRL B-30929. The recombinant construct was expressed in Escherichia coli BL21(DE3) with 6 × histidine at the C-terminus and the purified protein obtained as a single band of approx. 44 kDa on SDS-PAGE and 90 kDa on native-PAGE. The LbFDH was highly active at acidic conditions (pH 4.8–6.2). Its optimum temperature was 60 °C and 50 °C with NADP+ and NAD+, respectively and its Tm value was 78 °C. Its activity did not decrease after incubation in a solution containing 20% of DMSO and acetonitrile for 6 h. The KM constants were 49.8, 0.12 and 1.68 mM for formate (with NADP+), NADP+ and NAD+, respectively.


An NADP+ dependent FDH from L. buchneri NRRL B-30929 was cloned, expressed and identified with its unusual characteristics. The LbFDH can be a promising candidate for NADPH regeneration through biocatalysis requiring acidic conditions and high temperatures.


Acidic formate dehydrogenase Biochemical and kinetic characterization Highly NADP+ dependent formate dehydrogenase Lactobacillus buchneri NRRL B-30929 Solvent stable Thermostability 



This work was supported by Research Fund of the Yildiz Technical University (Project Number: FDK-2018-3331) and special thanks to Dr. Siqing Liu from USDA ARS for providing chromosomal DNA of Lactobacillus buchneri NRRL B-30929.

Supporting information

Supplementary Fig. 1—MALDI-TOF MS analysis of His6-LbFDH

Supplementary Fig. 2—The Differential Scanning Calorimetry (DSC) result of LbFDH

Supplementary Fig. 3—Effect of metal ions on the activity of recombinant LbFDH.

Supplementary Fig. 4—Organic solvent stability of LbFDH in 20% DMSO and acetonitrile for 6 h.

Supplementary Fig. 5—Effect of different storage temperatures on the stability of LbFDH.

Supplementary Fig. 6 a, b, c, d—Michaelis–Menten curves for NAD+ and NADP+ dependent reaction of LbFDH.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10529_2018_2568_MOESM1_ESM.docx (955 kb)
Supplementary material 1 (DOCX 933 kb)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Saadet Alpdağtaş
    • 1
    • 2
  • Sevil Yücel
    • 2
  • Handan Açelya Kapkaç
    • 3
  • Siqing Liu
    • 4
  • Barış Binay
    • 5
  1. 1.Department of Biology, Faculty of ScienceVan Yuzuncu Yil UniversityVanTurkey
  2. 2.Department of Bioengineering, Faculty of Chemistry and MetallurgyYildiz Technical UniversityIstanbulTurkey
  3. 3.Department of BiologyAnadolu UniversityEskişehirTurkey
  4. 4.U.S. Department of Agriculture, Renewable Product Technology Research UnitNational Centre for Agricultural Utilization ResearchPeoriaUSA
  5. 5.Department of BioengineeringGebze Technical UniversityKocaeliTurkey

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