Advertisement

Enzymatic Synthesis of Nucleoside Triphosphates and Deoxynucleoside Triphosphates by Surface-Displayed Kinases

  • Yi Ding
  • Ling OuEmail author
  • Qingbao DingEmail author
Article
First Online:
  • 57 Downloads

Abstract

Nucleoside triphosphates and deoxynucleoside triphosphates are important biochemical molecules. In this study, recombinant Escherichia coli that could display nucleotide kinases (INP-N-NMKases) and acetate kinase (INP-N-ACKase) on the cell surface were constructed by fusing an enzyme (NMKase/ACKase) to the N-terminus of ice nucleation protein (INP-N). By using intact recombinant bacteria cells as a catalyst coupled with an ACKase-catalyzed adenosine-5′-triphosphate (ATP) regeneration system, nucleoside triphosphates (NTPs) and deoxynucleoside triphosphates (dNTPs) could be synthesized efficiently. In a reaction system with 5 mmol/l substrate, the conversion rates of cytidine-5′-triphosphate (CTP) and deoxycytidine-5′-triphosphate (dCTP) were 96% and 93%, respectively, the conversion rate of ATP and deoxyadenosine-5′-triphosphate (dATP) was 96%, the conversion rate of deoxythymidine-5′-triphosphate (dTTP) was 91%, and the conversion rate of uridine-5′-triphosphate (UTP) was 80%. There was no obvious degradation. At 37 °C, the stability of the surface-displayed fusion protein, especially in the presence of the substrate, was significantly improved. Each whole cell could be reused more than 8 times.

Keywords

Nucleoside triphosphates Deoxynucleoside triphosphates Nucleotide kinase Acetate kinase Ice nucleation protein Surface display ATP regeneration 
This is a preview of subscription content, log in to check access.

Notes

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

12010_2019_3138_MOESM1_ESM.docx (444 kb)
ESM 1 (DOCX 444 kb)

References

  1. 1.
    Li, Y., Li, X. M., Zhang, H., Xu, R. F., & Tao, G. C. (2008). Effect of cytidine triphosphate and sodium ferulate on nerve conduction velocity in painful diabetic neuropathy. Journal of the Fourth Military Medical University, 22, 2075–2077.Google Scholar
  2. 2.
    Fang, L., Wang, J., & Zhao, Z. H. (2007). Curative effect of cytidine triphosphate via intramuscular injection in 30 patients with diabetic peripheral neuropathy. Journal of the Fourth Military Medical University, 28, 2275–2277.Google Scholar
  3. 3.
    Li, X. M., Li, Y., Zhao, K. Y., & Sun, L. H. (2005). Effect of cytidine triphosphate on nerve conduction velocity in patients with diabetic peripheral neuropathy. Chinese Journal of Clinical Rehabilitation, 9, 152–153.Google Scholar
  4. 4.
    Agteresch, H. J., Dagnelie, P. C., van den Berg, J. W., & Wilson, J. H. (1999). Adenosine triphosphate: Established and potential clinical applications. Drugs., 58(2), 211–232.CrossRefGoogle Scholar
  5. 5.
    Bennett, W. D., Zeman, K. L., Foy, C., Shaffer, C. L., Johnson, F. L., Regnis, J. A., Sannuti, A., & Johnson, J. (2001). Effect of aerosolized uridine 5′-triphosphate on mucociliary clearance in mild chronic bronchitis. American Journal of Respiratory and Critical Care Medicine, 164(2), 302–306.CrossRefGoogle Scholar
  6. 6.
    Sudo, E., Lee, M. M., Boyd, W. A., & King, M. (2000). Effects of methacholine and uridine 5′-triphosphate on tracheal mucus rheology in mice. American Journal of Respiratory Cell and Molecular Biology, 22(3), 373–379.CrossRefGoogle Scholar
  7. 7.
    Wihlborg, A. K., Balogh, J., Wang, L., Borna, C., Dou, Y., Joshi, B. V., Lazarowski, E., Jacobson, K. A., Arner, A., & Erlinge, D. (2006). Positive inotropic effects by uridine triphosphate (UTP) and uridine diphosphate (UDP) via P2Y2 and P2Y6 receptors on cardiomyocytes and release of UTP in man during myocardial infarction. Circulation Research, 98(7), 970–976.CrossRefGoogle Scholar
  8. 8.
    Erlinge, D., Harnek, J., van Heusden, C., Olivecrona, G., Jern, S., & Lazarowski, E. (2005). Uridine triphosphate (UTP) is released during cardiac ischemia. International Journal of Cardiology, 100(3), 427–433.CrossRefGoogle Scholar
  9. 9.
    Antoun, A., Pavlov, M. Y., Andersson, K., Tenson, T., & Ehrenberg, M. (2003). The roles of initiation factor 2 and guanosine triphosphate in initiation of protein synthesis. The EMBO Journal, 22(20), 5593–5601.CrossRefGoogle Scholar
  10. 10.
    Menkes, D. B., Rasenick, M. M., Wheeler, M. A., & Bitensky, M. W. (1983). Guanosine triphosphate activation of brain adenylate cyclase: Enhancement by long-term antidepressant treatment. Science., 219(4580), 65–67.CrossRefGoogle Scholar
  11. 11.
    Hocek, M. (2019). Enzymatic synthesis of base-functionalized nucleic acids for sensing, cross-linking, and modulation of protein-DNA binding and transcription. Accounts of Chemical Research, 52, 1730–1737.CrossRefGoogle Scholar
  12. 12.
    Marie, F., Luke, K. M., Ivo, S., & Marcel, H. (2019). Chemical methods for the modification of RNA. Methods., 161, 64–82.CrossRefGoogle Scholar
  13. 13.
    Fernandez-Lucas, J. (2015). Multienzymatic synthesis of nucleic acid derivatives: A general perspective. Applied Microbiology and Biotechnology, 99(11), 4615–4627.CrossRefGoogle Scholar
  14. 14.
    Yano, T., Yasohara, Y., Kashihara, M., Tachiki, T., Hidehiko, K., & Tatsurokuro, T. (1989). Production of deoxyribonucleoside triphosphates through coupled fermentation with energy transfer. Agricultural and Biological Chemistry, 53, 1935–1940.Google Scholar
  15. 15.
    You, Y., Ding, Q. B., & Ou, L. (2007). Biosynthesis of nucleosides triphosphate by immobilized beer yeast cells. Industrial Microbiology., 37, 31–35.Google Scholar
  16. 16.
    Wu, W., Bergstrom, D. E., & Davisson, V. J. (2003). A combination chemical and enzymatic approach for the preparation of azole carboxamide nucleoside triphosphate. The Journal of Organic Chemistry, 68(10), 3860–3865.CrossRefGoogle Scholar
  17. 17.
    Bochkov, D. V., Khomov, V. V., & Tolstikova, T. G. (2006). Hydrolytic approach for production of deoxyribonucleoside-and ribonucleoside-5′-monophosphates and enzymatic synthesis of their polyphosphates. Biochemistry (Moscow), 71(1), 79–83.CrossRefGoogle Scholar
  18. 18.
    Scott, L. G., Geierstanger, B. H., Williamson, J. R., & Hennig, M. (2004). Enzymatic synthesis and 19F NMR studies of 2-fluoroadenine-substituted RNA. Journal of the American Chemical Society, 126(38), 11776–11777.CrossRefGoogle Scholar
  19. 19.
    Lee, J. H., Chung, S. W., Lee, H. J., Jang, K. S., Lee, S. G., & Kim, B. G. (2010). Optimization of the enzymatic one pot reaction for the synthesis of uridine 5′-diphosphogalactose. Bioprocess and Biosystems Engineering, 33(1), 71–78.CrossRefGoogle Scholar
  20. 20.
    Becker, S., Hobenreich, H., Vogel, A., Knorr, J., Wilhelm, S., Rosenau, F., Jaeger, K. E., Reetz, M. T., & Kolmar, H. (2008). Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes. Angewandte Chemie (International Ed. in English), 47(27), 5085–5088.CrossRefGoogle Scholar
  21. 21.
    Becker, S., Michalczyk, A., Wilhelm, S., Jaeger, K. E., & Kolmar, H. (2007). Ultrahigh-throughput screening to identify Escherichia coli cells expressing functionally active enzymes on their surface. Chembiochem., 8(8), 943–949.CrossRefGoogle Scholar
  22. 22.
    Mazor, Y., van Blarcom, T., Iverson, B. L., & Georgiou, G. (2008). E-clonal antibodies: Selection of full-length IgG antibodies using bacterial periplasmic display. Nature Protocols, 3(11), 1766–1777.CrossRefGoogle Scholar
  23. 23.
    Jeiranikhameneh, M., Razavi, M. R., Irani, S., Siadat, S. D., & Oloomi, M. (2017). Designing novel construction for cell surface display of protein E on Escherichia coli using non-classical pathway based on Lpp-OmpA. AMB Express, 7(1), 53.CrossRefGoogle Scholar
  24. 24.
    Maruthamuthu, M. K., Selvamani, V., Nadarajan, S. P., Yun, H., Oh, Y. K., Eom, G. T., & Hong, S. H. (2018). Manganese and cobalt recovery by surface display of metal binding peptide on various loops of OmpC in Escherichia coli. Journal of Industrial Microbiology & Biotechnology, 45(1), 31–41.CrossRefGoogle Scholar
  25. 25.
    van Bloois, E., Winter, R. T., Kolmar, H., & Fraaije, M. W. (2011). Decorating microbes: Surface display of proteins on Escherichia coli. Trends in Biotechnology, 29(2), 79–86.CrossRefGoogle Scholar
  26. 26.
    van Bloois, E., Winter, R. T., Janssen, D. B., & Fraaije, M. W. (2009). Export of functional streptomyces coelicolor alditol oxidase to the periplasm or cell surface of Escherichia coli and its application in whole-cell biocatalysis. Applied Microbiology and Biotechnology, 83(4), 679–687.CrossRefGoogle Scholar
  27. 27.
    Cho, C. M., Mulchandani, A., & Chen, W. (2002). Bacterial cell surface display of organophosphorus hydrolase for selective screening of improved hydrolysis of organophosphate nerve agents. Applied and Environmental Microbiology, 68(4), 2026–2030.CrossRefGoogle Scholar
  28. 28.
    Bao, S., Yu, S., Guo, X., Zhang, F., Sun, Y., Tan, L., Duan, Y., Lu, F., Qiu, X., & Ding, C. (2015). Construction of a cell-surface display system based on the N-terminal domain of ice nucleation protein and its application in identification of mycoplasma adhesion proteins. Journal of Applied Microbiology, 119(1), 236–244.CrossRefGoogle Scholar
  29. 29.
    Kang, D. G., Li, L., Ha, J. H., Choi, S. S., & Cha, H. J. (2008). Efficient cell surface display of organophosphorous hydrolase using N-terminal domain of ice nucleation protein in Escherichia coli. Korean Journal of Chemical Engineering, 25(4), 804–807.CrossRefGoogle Scholar
  30. 30.
    Li, L., Kang, D. G., & Cha, H. J. (2004). Functional display of foreign protein on surface of Escherichia coli using N-terminal domain of ice nucleation protein. Biotechnology and Bioengineering, 85(2), 214–221.CrossRefGoogle Scholar
  31. 31.
    Yao, Y., Ding, Q., & Ou, L. (2019). Biosynthesis of (deoxy)guanosine-5′-triphosphate by GMP kinase and acetate kinase fixed on the surface of Escherichia coli. Enzyme and Microbial Technology, 122, 82–89.CrossRefGoogle Scholar
  32. 32.
    Devedjiev, I. T. (2006). A novel method for the synthesis of acetyl phosphate. Phosphorus, Sulfur and Silicon and the Related Elements, 181(8), 1785–1787.CrossRefGoogle Scholar
  33. 33.
    Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual (2nd ed.). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.Google Scholar
  34. 34.
    Reynes, J. P., Tiraby, M., Baron, M., Drocourt, D., & Tiraby, G. (1996). Escherichia coli thymidylate kinase: Molecular cloning, nucleotide sequence, and genetic organization of the corresponding tmk locus. Journal of Bacteriology, 178(10), 2804–2812.CrossRefGoogle Scholar
  35. 35.
    Yan, B., Ding, Q., Ou, L., & Zou, Z. (2014). Production of glucose-6-phosphate by glucokinase coupled with an ATP regeneration system. World Journal of Microbiology and Biotechnology, 30(3), 1123–1128.CrossRefGoogle Scholar
  36. 36.
    Zhang, J., Qian, Y., Ding, Q., & Ou, L. (2015). Enzymatic manufacture of deoxythymidine-5′-triphosphate with permeable intact cells of Escherichia coli coexpressing thymidylate kinase and acetate kinase. Journal of Microbiology and Biotechnology, 25(12), 2034–2042.CrossRefGoogle Scholar
  37. 37.
    Liu, Y., Wang, J., Xu, C., Chen, Y., Yang, J., Liu, D., Niu, H., Jiang, Y., Yang, S., & Ying, H. (2016). Efficient multi-enzyme-catalyzed CDP-choline production driven by an ATP donor module. Applied Microbiology and Biotechnology, 101, 1409–1417.CrossRefGoogle Scholar
  38. 38.
    Ishige, K., Hamamoto, T., Shiba, T., & Noguchi, T. (2001). Novel method for enzymatic synthesis of CMP-NeuAc. Bioscience, Biotechnology, and Biochemistry, 65(8), 1736–1740.CrossRefGoogle Scholar
  39. 39.
    Zhang, J., Wu, B., Zhang, Y., Kowal, P., & Wang, P. G. (2003). Creatine phosphate--creatine kinase in enzymatic synthesis of glycoconjugates. Organic Letters, 5(15), 2583–2586.CrossRefGoogle Scholar
  40. 40.
    Resnick, S. M., & Zehnder, A. J. (2000). In vitro ATP regeneration from polyphosphate and AMP by polyphosphate: AMP phosphotransferase and adenylate kinase from acinetobacter johnsonii 210A. Applied and Environmental Microbiology, 66(5), 2045–2051.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiChina
  2. 2.Department of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical SchoolRutgers UniversityNewarkUSA

Personalised recommendations

Cite article

Buy options