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Replacement P212H Altered the pH–Temperature Profile of Phytase from Aspergillus niger NII 08121

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Abstract

Microbial phytase, a widely used animal feed enzyme, needs to be active and stable in the acidic milieu for better performance in the monogastric gut. Aspergillus niger phytases exhibit an activity dip in the pH range from 3.0 to 3.5. Replacement of amino acids, which changed the pKa of catalytic residues H82 and D362, resulted in alteration of the pH profile of a thermostable phytase from A. niger NII 08121. Substitution P212H in the protein loop at 14 Å distance to the active site amended the pH optimum from 2.5 to pH 3.2 nevertheless with a decrease in thermostability than the wild enzyme. This study described the utility of amino acid replacements based on pKa shifts of catalytic acid/base to modulate the pH profile of phytases.

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References

  1. Pandey, A., Szakacs, G., Soccol, C. R., Rodriguez-Leon, J. A., & Soccol, V. T. (2001). Bioresource Technology, 77, 203–214.

    Article  CAS  Google Scholar 

  2. Vats, P., & Banerjee, U. C. (2004). Enzyme and Microbial Technology, 35, 3–14.

    Article  CAS  Google Scholar 

  3. Yi, Z., & Kornegay, E. T. (1996). Animal Feed Science and Technology, 61, 361–368.

    Article  CAS  Google Scholar 

  4. Ullah, A. H. J., & Gibson, D. M. (1987). Preparative Biochemistry, 17, 63–91.

    Article  CAS  Google Scholar 

  5. Ushasree, M. V., Vidya, J., & Pandey, A. (2014). Process Biochemistry, 49, 1440–1447.

    Article  CAS  Google Scholar 

  6. Augspurger, N. R., & Baker, D. H. (2004). Journal of Animal Sciences, 82, 1100–1107.

    CAS  Google Scholar 

  7. Joshi, M. D., Sidhu, G., Nielsen, J. E., Brayer, G. D., Withers, S. G., & McIntosh, L. P. (2001). Biochemistry, 40, 10115–10139.

    Article  CAS  Google Scholar 

  8. Cockburn, D. W., & Clarke, A. J. (2011). Protein Engineering Design and Selection, 24, 429–437.

    Article  CAS  Google Scholar 

  9. Tomschy, A., Brugger, R., Lehmann, M., Svendsen, A., Vogel, K., Kostrewa, D., et al. (2002). Applied Environmental Microbiology, 68, 1907–1913.

    Article  CAS  Google Scholar 

  10. Kim, T., Mullaney, E. J., Porres, J. M., Roneker, K. R., Crowe, S., Rice, S., et al. (2006). Applied Environmental Microbiology, 72, 4397–4403.

    Article  CAS  Google Scholar 

  11. Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., et al. (2007). Bioinformatics, 23, 2947–2948.

    Article  CAS  Google Scholar 

  12. Petersen, B., Petersen, T. N., Andersen, P., Nielsen, M., & Lundegaard, C. (2009). BMC Structural Biology, 9, 6807–6809.

    Article  Google Scholar 

  13. Bas, D. C., Rogers, D. M., & Jensen, J. H. (2008). Proteins, 73, 765–783.

    Article  CAS  Google Scholar 

  14. Guex, N., & Peitsch, M. C. (1997). Electrophoresis, 18, 2714–2723.

    Article  CAS  Google Scholar 

  15. Lin-Cereghino, J., Wong, W. W., Xiong, S., Giang, W., Luong, L. T., Vu, J., et al. (2005). Biotechniques, 38, 44–48.

    Article  CAS  Google Scholar 

  16. Heinonen, J. K., & Lathi, R. J. (1981). Analytical Biochemistry, 113, 313–317.

    Article  CAS  Google Scholar 

  17. Cheng, J., Randall, A., & Baldi, P. (2006). Proteins, 62, 1125–1132.

    Article  CAS  Google Scholar 

  18. Capriotti, E., Fariselli, P., & Casadio, R. (2005). Nucleic Acid Research, 33, 306–310.

    Article  Google Scholar 

  19. Yin, S., Ding, F., & Dokholyan, N. V. (2007). Nature Methods, 4, 466–467.

    Article  CAS  Google Scholar 

  20. Ushasree, M. V., Vidya, J., & Pandey, A. (2014). Biotechnology Letters, 36, 85–91.

    Article  CAS  Google Scholar 

  21. Kostrewa, D., Grueninger-Leitch, F., D'Arcy, A., Broger, C., Mitchell, D., Adolphus, P. G. M., et al. (1997). Nature Structural Biology, 4, 185–190.

    Article  CAS  Google Scholar 

  22. Shimizu, M. (1993). Bioscience. Biotechnology and Biochemistry, 57, 1364–1365.

    Article  CAS  Google Scholar 

  23. Pasamontes, L., Haiker, M., Wyss, M., Tessier, M., & van Loon, A. P. (1997). Applied Environmental Microbiology, 63, 1696–1700.

    CAS  Google Scholar 

  24. Ullah, A. H. J., & Sethumadhavan, K. (2003). Biochemical and Biophysical Research Communication., 303, 463–468.

    Article  CAS  Google Scholar 

  25. Rodriguez, E., Han, Y., & Lei, X. G. (1999). Biochemical and Biophysical Research Communication, 257, 117–123.

    Article  CAS  Google Scholar 

  26. Mullaney, E. J., Daly, C. B., Kim, T., Porres, J. M., Lei, X. G., & Sethumadhavan, K. (2002). Biochemical and Biophysical Research Communication, 297, 1016–1020.

    Article  CAS  Google Scholar 

  27. Zhou, C., Xue, Y., & Ma, Y. (2010). Journal of Bioscience and Bioengineering, 110, 12–17.

    Article  CAS  Google Scholar 

  28. Thiltgen, G., & Goldstein, R. A. (2012). PLoS ONE. doi:10.1371/ 0046084.

    Google Scholar 

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Acknowledgments

One of the authors (MVU) thank the Indian Council of Medical Research, New Delhi for the award of a Senior Research Fellowship to her. Financial support from the Department of Biotechnology, New Delhi (Grant No. GAP 1410) is gratefully acknowledged.

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Authors and Affiliations

  1. Biotechnology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Trivandrum, 695 019, India

    Mrudula Vasudevan Ushasree, Jalaja Vidya & Ashok Pandey

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  1. Mrudula Vasudevan Ushasree
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  2. Jalaja Vidya
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  3. Ashok Pandey
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Correspondence to Ashok Pandey.

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Ushasree, M.V., Vidya, J. & Pandey, A. Replacement P212H Altered the pH–Temperature Profile of Phytase from Aspergillus niger NII 08121. Appl Biochem Biotechnol 175, 3084–3092 (2015). https://doi.org/10.1007/s12010-015-1485-6

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  • DOI: https://doi.org/10.1007/s12010-015-1485-6

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