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Microfluidic Multiple Cell Chip Reactor Filled with Enzyme-Coated Magnetic Nanoparticles — An Efficient and Flexible Novel Tool for Enzyme Catalyzed Biotransformations

Journal of Flow Chemistry volume 6, pages 43–52 (2016)Cite this article

Abstract

Biotransformation of L-phenylalanine (L-1a) and five unnatural substrates (rac-1b–f) by phenylalanine ammonia-lyase (PAL) was investigated in a novel microfluidic device (Magne-Chip) that comprises microliter volume reaction cells filled with PAL-coated magnetic nanoparticles (MNPs). Experiments proved the excellent reproducibility of enzymecatalyzed biotransformation in the chip and the excellent reusability of the enzyme layer during 14 h continuous measurement (>98% over 7 repetitive measurements with L-1a). The platform also enabled fully automatic multiparameter measurements with a single biocatalyst loading of about 1 mg PAL-MNP. Computational fluid dynamics (CFD) calculations were used to study the flow field in the chambers and the effect of unintended bubble formation. Optimal flow rate for L-1a reaction and specific activities for rac-1b–f under these conditions were determined.

References

  1. Poppe, L.; Paizs, C.; Kovacs, K.; Irimie, F. D.; Vértessy, B. G. Meth. Mol. Biol. 2012, 794, 3–19.

    CAS  Article  Google Scholar 

  2. Hodgins, D.S. J. Biol. Chem. 1971, 246, 2977–2985.

    CAS  Google Scholar 

  3. Sarkissian, C. N.; Gamez, A. Mol. Gen. Metab. 2005, 86, 22–26.

    Article  Google Scholar 

  4. Hughes, A. Amino Acids, Peptides and Proteins in Organic Chemistry Volume 1; Weinheim Germany: Wiley VCH, 2009; p. 94. ISBN 9783527320967.

    Google Scholar 

  5. Munirathinam, R.; Huskens, J.; Verboom, W. Adv. Synth. Catal. 2015, 357, 1093–1123.

    CAS  Article  Google Scholar 

  6. Sheng, J.; Zhang, L.; Lei, J.; Ju, H. Anal. Chim. Acta 2012, 709, 41–46.

    CAS  Article  Google Scholar 

  7. Wang, M. S.; Black, J. C.; Knowles, M. K.; Reed, S. M. Anal. Bioanal. Chem. 2011, 401, 1309–1318.

    CAS  Article  Google Scholar 

  8. Song, Y. S.; Shin, H. Y.; Lee, J. Y.; Park, C.; Kim, S. W. Food Chem. 2012, 133, 611–617.

    CAS  Article  Google Scholar 

  9. He, P.; Greenway, G.; Haswell, S. J. Microfluid. Nanofluid. 2010, 8, 565–573.

    CAS  Article  Google Scholar 

  10. Gijs, M. A. M.; Lacharme, F.; Lehmann, U. Chem. Rev. 2010,110, 1518–1563.

    CAS  Article  Google Scholar 

  11. Weiser, D.; Bencze, L. C.; Banóczi, G.; Ender, F.; Kókai, E.; Szilagyi, A.; Vértessy, B. G.; Farkas, Ö; Paizs, C.; Poppe, L. ChemBioChem 2015, 16, 2283–2288.

    CAS  Article  Google Scholar 

  12. Ender, F.; Weiser, D.; Vitéz, A.; Sallai, G.; Németh, M.; Poppe, L. Microsyst. Technol. [First online: 14 December 2015] DOI 10.1007/s00542- 015-2749-3.

    Google Scholar 

  13. ANSYS Fluent, ANSYS Ver 16.0 (ANSYS Inc. Canonsburg, USA).

  14. Lilly, M. D.; Hornby, W. E.; Crook, E. M. Biochem. J 1966, 100, 718–723.

    CAS  Article  Google Scholar 

  15. Paizs, C.; Katona, A.; Rétey, J. Chem. Eur J. 2006, 12, 2739–2744.

    CAS  Article  Google Scholar 

  16. Gloge, A.; Zon, J.; Kovari, Â.; Poppe, L.; Rétey, J. Chem. Eur. J. 2000, 6, 3386–3390.

    CAS  Article  Google Scholar 

  17. Bradford, M. M. Anal. Biochem. 1976, 72, 248–254.

    CAS  Article  Google Scholar 

Download references

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Authors and Affiliations

  1. Department of Electron Devices, Budapest University of Technology and Economics, Budapest, Magyar tudósok körútja 2, H-1117, Budapest, Hungary

    Ferenc Ender & Péter Pálovics

  2. Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, Műegyetem rkp. 3, H-1111, Budapest, Hungary

    Diána Weiser & László Poppe

  3. Biocatalysis and Biotransformation Research Group, Babeş-Bolyai University of Cluj-Napoca, Arany János str. 11, Ro-400028, Cluj-Napoca, Romania

    Botond Nagy, Csaba László Bencze & Csaba Paizs

  4. SynBiocat Ltd., H-1173, Budapest, Lázár deák u. 4/1., Hungary

    László Poppe

Authors
  1. Ferenc Ender
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  2. Diána Weiser
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  3. Botond Nagy
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  4. Csaba László Bencze
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  5. Csaba Paizs
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  6. Péter Pálovics
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  7. László Poppe
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Correspondence to Ferenc Ender or László Poppe.

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Ender, F., Weiser, D., Nagy, B. et al. Microfluidic Multiple Cell Chip Reactor Filled with Enzyme-Coated Magnetic Nanoparticles — An Efficient and Flexible Novel Tool for Enzyme Catalyzed Biotransformations. J Flow Chem 6, 43–52 (2016). https://doi.org/10.1556/1846.2015.00036

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  • DOI: https://doi.org/10.1556/1846.2015.00036

Keyword

  • magnetic nanoparticle
  • magnetic chip reactor
  • continuous-flow biotransformation
  • phenylalanine ammonia-lyase
  • unnatural amino acid