Applied Microbiology and Biotechnology

, Volume 65, Issue 1, pp 110–118 | Cite as

Molecular and physiological characterisation of a 3-phytase from soil bacterium Klebsiella sp. ASR1

  • A. Sajidan
  • A. Farouk
  • R. Greiner
  • P. Jungblut
  • E.-C. Müller
  • R. BorrissEmail author
Original Paper


Klebsiella sp. strain ASR1 isolated from an Indonesian rice field is able to hydrolyse myo-inositol hexakis phosphate (phytate). The phytase protein was purified and characterised as a 42 kDa protein accepting phytate, NADP and sugar phosphates as substrates. The corresponding gene (phyK) was cloned from chromosomal DNA using a combined approach of protein and genome analysis, and expressed in Escherichia coli. The recombinant enzyme was identified as a 3-phytase yielding myo-inositol monophosphate, Ins(2)P, as the final product of enzymatic phytate hydrolysis. Based on its amino acid sequence, PhyK appears to be a member of a hitherto unknown subfamily of histidine acid phytate-degrading enzymes with the active site RHGXRXP and HD sequence motifs, and is different from other general phosphatases and phytases. Due to its ability to degrade sodium phytate to the mono phosphate ester, the phyK gene product is an interesting candidate for industrial and agricultural applications to make phytate phosphorous available for plant and animal nutrition.


Phytase Activity Phytase Gene Recombinant Phytase Histidine Acid Phosphatase pelB Signal Peptide 
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.



Financial support for A. S. from DAAD is gratefully acknowledged. We are especially grateful to Monika Schmid for analysis of peptide masses and Thomas Leya for his help in using the PAUP package for construction of evolutionary trees. We thank Romy Scholz and Kristin Rosner for their support in DNA sequence analysis. Dr. Steffen Porwollik is thanked for critical reading of the manuscript. The technical assistance of Christiane Müller and Sybille Striegl is gratefully acknowledged.

Supplementary material

Supplementary Material

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • A. Sajidan
    • 1
  • A. Farouk
    • 1
  • R. Greiner
    • 2
  • P. Jungblut
    • 3
  • E.-C. Müller
    • 3
  • R. Borriss
    • 1
    Email author
  1. 1.Department of Bacterial Genetics, Institute of BiologyHumboldt UniversityBerlinGermany
  2. 2.Centre for Molecular BiologyFederal Research Centre for NutritionKarlsruheGermany
  3. 3.Max Delbrück Centre for Molecular MedicineBerlinGermany

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