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Crystal structures of Escherichia coli phytase and its complex with phytate

Nature Structural Biology volume 7, pages 108113 (2000) | Download Citation

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Abstract

Phytases catalyze the hydrolysis of phytate and are able to improve the nutritional quality of phytate-rich diets. Escherichia coli phytase, a member of the histidine acid phosphatase family has the highest specific activity of all phytases characterized. The crystal structure of E. coli phytase has been determined by a two-wavelength anomalous diffraction method using the exceptionally strong anomalous scattering of tungsten. Despite a lack of sequence similarity, the structure closely resembles the overall fold of other histidine acid phosphatases. The structure of E. coli phytase in complex with phytate, the preferred substrate, reveals the binding mode and substrate recognition. The binding is also accompanied by conformational changes which suggest that substrate binding enhances catalysis by increasing the acidity of the general acid.

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Acknowledgements

The authors would like to thank Y.-C. Liou of P. Davies' group at Queen's University for equipment and technical assistance in early purification attempts and for his support throughout the project. We thank A. Tocilj for his encouragement and assistance at various stages of the project. We thank A. Iyo and M. Cotrill for constructing and help with purifying the inactive mutant, respectively. We also thank E. Leinala for her advice on crystallization. G. Thatcher provided helpful discussion. We are grateful to L. Flaks and the technical staff at the National Synchrotron Light Source at Brookhaven National Laboratory for their support at X8-C. D.L. was an Ontario Graduate Scholarship recipient. This work was supported by a NSERC grant to Z.J. and contract funding from Ontario Pork to C.W.F.

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  1. Department of Biochemistry, Queen's University, Kingston, Ontario K7L 3N6, Canada.

    • Daniel Lim
    •  & Zongchao Jia
  2. Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.

    • Serguei Golovan
    •  & Cecil W. Forsberg

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Correspondence to Zongchao Jia.

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https://doi.org/10.1038/72371