Skip to main content
Springer Nature Link
Log in

Engineering Fructosyl Peptide Oxidase to Improve Activity Toward the Fructosyl Hexapeptide Standard for HbA1c Measurement

  • Research
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

The recently discovered fructosyl peptide oxidase from Phaeosphaeria nodorum (PnFPOX) was demonstrated to react with the glycated hexapeptide measurement standard of hemoglobin A1c, fVHLTPE. The highly reactive Coniochaeta FPOX (FPOX-C) showed no detectable activity with the hexapeptide. Two loop regions were identified as having important effects on the enzymatic properties of FPOX. The first loop has a strong influence on the ability to bind larger glycated peptides, while the second loop has a significant effect on catalytic activity. Loop-substitution mutants showed that the highest activity against fVHLTPE resulted from the combination of the first loop from PnFPOX and the second loop from FPOX-C. The most promising engineered FPOX created, which showed 17-fold greater dehydrogenase activity against fVHLTPE than wild-type PnFPOX, was the FPOX-C mutant with a PnFPOX-derived loop 1 region and an Asn56Ala substitution.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  1. The Diabetes Control and Complications Trial Research Group. (1993). The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. New England Journal of Medicine, 329, 977–986.

    Article  Google Scholar 

  2. The Diabetes Control and Complications Trial Research Group. (1996). Influence of intensive diabetes treatment on quality-of-life outcomes. Diabetes Care, 19, 195–203.

    Article  Google Scholar 

  3. American Diabetes Association. (2012). Position statement: Standards of medical care in diabetes. Diabetes Care, 35, S11–S63.

    Article  Google Scholar 

  4. Ferri, S., Kim, S., Tsugawa, W., & Sode, K. (2009). Review of fructosyl amino acid oxidase engineering research: A glimpse into the future of hemoglobin A1c biosensing. Journal of Diabetes Science and Technology, 3, 585–592.

    Google Scholar 

  5. Jeppsson, J. O., Kobold, U., Barr, J., Finke, A., Hoelzel, W., Hoshino, T., et al. (2002). Approved IFCC Reference Method for the measurement of HbA1c in human blood. Clinical Chemistry and Laboratory Medicine, 40, 78–89.

    Article  CAS  Google Scholar 

  6. Horiuchi, T., & Kurokawa, T. (1005). (1991) Purification and properties of fructosylamine oxidase from Aspergillus sp. Agricultural and Biological Chemistry, 55(2), 333–338.

    Article  Google Scholar 

  7. Yoshida, N., Sakai, Y., Serata, M., Tani, Y., & Kato, N. (1995). Distribution and properties of fructosyl amino acid oxidase in fungi. Applied and Environment Microbiology, 61(12), 4487–4489.

    CAS  Google Scholar 

  8. Sode, K., Ishimura, F., & Tsugawa, W. (2001). Screening and characterization of fructosyl-valine-utilizing marine microorganisms. Marine biotechnology (NY), 3(2), 126–132.

    Article  CAS  Google Scholar 

  9. Horiuchi, T., Kurokawa, T., & Saito, N. (1989). Purification and properties of fructosyl-amino acid oxidase from Corynebacterium sp. 2-4-1. Agricultural and Biological Chemistry, 53(1), 103–110.

    Article  CAS  Google Scholar 

  10. Ferri, S., Sakaguchi, A., Goto, H., Tsugawa, W., & Sode, K. (2005). Isolation and characterization of a fructosyl-amine oxidase from an Arthrobacter sp. Biotechnology Letters, 27, 27–32.

    Article  CAS  Google Scholar 

  11. Miura, S., Ferri, S., Tsugawa, W., Kim, S., & Sode, K. (2006). Active site analysis of fructosyl amine oxidase using homology modeling and site-directed mutagenesis. Biotechnology Letters, 28, 1895–1900.

    Article  CAS  Google Scholar 

  12. Fujiwara, M., Sumitani, J., Koga, S., Yoshioka, I., Kouzuma, T., et al. (2006). Alteration of substrate specificity of fructosyl-amino acid oxidase from Ulocladium sp. JS-103. Journal of Bioscience and Bioengineering, 102, 241–243.

    Article  CAS  Google Scholar 

  13. Fujiwara, M., Sumitani, J., Koga, S., Yoshioka, I., Kouzuma, T., et al. (2007). Alteration of substrate specificity of fructosyl-amino acid oxidase from Fusarium oxysporum. Applied Microbiology and Biotechnology, 74, 813–819.

    Article  CAS  Google Scholar 

  14. Miura, S., Ferri, S., Tsugawa, W., Kim, S., & Sode, K. (2008). Development of fructosyl amine oxidase specific to fructosyl valine by site-directed mutagenesis. Protein Engineering, Design & Selection, 21, 233–239.

    Article  CAS  Google Scholar 

  15. Hirokawa, K., Ichiyanagi, A., & Kajiyama, N. (2008). Enhancement of thermostability of fungal deglycating enzymes by directed evolution. Applied Microbiology and Biotechnology, 78, 775–781.

    Article  CAS  Google Scholar 

  16. Kim, S., Nibe, E., Ferri, S., Tsugawa, W., & Sode, K. (2010). Engineering of dye-mediated dehydrogenase property of fructosyl amino acid oxidases by site-directed mutagenesis studies of its putative proton relay system. Biotechnology Letters, 32(8), 1123–1129.

    Article  CAS  Google Scholar 

  17. Zheng, J., Guan, H., Xu, L., Yang, R., & Lin, Z. (2010). Engineered amadoriase II exhibiting expanded substrate range. Applied Microbiology and Biotechnology, 86, 607–613.

    Article  CAS  Google Scholar 

  18. Kim, S., Nibe, E., Tsugawa, W., Kojima, K., Ferri, S., & Sode, K. (2012). Construction of engineered fructosyl peptidyl oxidase for enzyme sensor applications under normal atmospheric conditions. Biotechnology Letters, 34(3), 491–497.

    Article  CAS  Google Scholar 

  19. Hirokawa, K., Gomi, K., & Kajiyama, N. (2003). Molecular cloning and expression of novel fructosyl peptide oxidases and their application for the measurement of glycated protein. Biochemical and Biophysical Research Communications, 311, 104–111.

    Article  CAS  Google Scholar 

  20. Kim, S., Ferri, S., Tsugawa, W., Mori, K., & Sode, K. (2010). Motif-based search for a novel fructosyl peptide oxidase from genome databases. Biotechnology and Bioengineering, 106, 358–366.

    Article  CAS  Google Scholar 

  21. Finot, P. A., Viani, R., Bricout, J., & Mauron, J. (1969). Detection and identification of pyridoxine, a second lysine derivative obtained upon acid hydrolysis of heated milk. Experientia, 25, 134–135.

    Article  CAS  Google Scholar 

  22. Baker, J. R., Zyzak, D. V., Thorpe, S. R., & Baynes, J. W. (1993). Mechanism of fructosamine assay: Evidence against role of superoxide as intermediate in nitroblue tetrazolium reduction. Clinical Chemistry, 39, 2460–2465.

    CAS  Google Scholar 

  23. Collard, F., Zhang, J., Nemet, I., Qanungo, K. R., Monnier, V. M., & Yee, V. C. (2008). Crystal structure of the deglycating enzyme fructosamine oxidase (Amadoriase II). Journal of Biological Chemistry, 283, 27007–27016.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei-shi, Tokyo, 184-8588, Japan

    Stefano Ferri, Yusuke Miyamoto, Wakako Tsugawa & Koji Sode

  2. School of Biotechnology and Bioscience, Tokyo University of Technology, Hachioji, Japan

    Akane Sakaguchi-Mikami

Authors
  1. Stefano Ferri
    View author publications

    Search author on:PubMed Google Scholar

  2. Yusuke Miyamoto
    View author publications

    Search author on:PubMed Google Scholar

  3. Akane Sakaguchi-Mikami
    View author publications

    Search author on:PubMed Google Scholar

  4. Wakako Tsugawa
    View author publications

    Search author on:PubMed Google Scholar

  5. Koji Sode
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Koji Sode.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ferri, S., Miyamoto, Y., Sakaguchi-Mikami, A. et al. Engineering Fructosyl Peptide Oxidase to Improve Activity Toward the Fructosyl Hexapeptide Standard for HbA1c Measurement. Mol Biotechnol 54, 939–943 (2013). https://doi.org/10.1007/s12033-012-9644-2

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12033-012-9644-2

Keywords

Profiles

  1. Koji Sode View author profile