MIO-enzyme toolbox: cloning, expression and purification of recombinant „Rt”PAL

Authors

  • Alina FILIP Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro
  • László Csaba BENCZE Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro
  • Csaba PAIZS Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro https://orcid.org/0000-0002-7403-7098
  • László POPPE Department of Organic Chemistry and Technology, Budapest University of Technology and Economics. E-mail: irimie@chem.ubbcluj.ro https://orcid.org/0000-0002-8358-1378
  • Florin Dan IRIMIE Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro https://orcid.org/0000-0003-2093-0136

Keywords:

MIO–enzyme, non-natural amino acid, phenylalanine ammonia-lyase, phenylketonuria.

Abstract

The recent advances in the molecular engineering of aromatic ammonia lyases (ALs) and aminomutases (AMs) attracted increased interest towards their applications in the treatment of phenylketonuria and/or the synthesis of non-natural amino acids. Herein we describe the cloning, isolation and purification of recombinant L-phenylalanine ammonia-lyase from Rhodosporidium toruloides for future biocatalytic and therapeutic applications.

Author Biographies

Alina FILIP, Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

László Csaba BENCZE, Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

Csaba PAIZS, Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

László POPPE, Department of Organic Chemistry and Technology, Budapest University of Technology and Economics. E-mail: irimie@chem.ubbcluj.ro

Department of Organic Chemistry and Technology, Budapest University of Technology and Economics. E-mail: irimie@chem.ubbcluj.ro

Florin Dan IRIMIE, Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

Biocatalysis and Biotransformation Research Group, Department of Chemistry, Babeş-Bolyai University, Cluj-Napoca, Romania. E-mail: irimie@chem.ubbcluj.ro

References

Babich, O. O., Pokrovsky, V. S., Anisimova, N. Y., Sokolov, N. N., Prosekov, A. Y. (2013) Recombinant l-phenylalanine ammonia lyase from Rhodosporidium toruloides as a potential anticancer agent: l -Phenylalanine Ammonia Lyase Аnticancer Activity, Biotechnol. Appl. Bioc., 60, 316–322

Bánóczi, G., Szabó, C., Bata, Z., Filip, A., Hornyánszky, G., Poppe, L. (2015) Structural modeling of phenylalanine ammonia-lyases and related MIO-containing enzymes – An insight into thermostability and ionic interactions, Studia UBB Chemia, vol. 4, 213–228

Gamez, A., Sarkissian, C., Wang, L., Kim, W., Straub, M., Patch, M., Chen, L., Striepeke, S., Fitzpatrick, P., Lemontt, J. (2005) Development of pegylated forms of recombinant phenylalanine ammonia-lyase for the treatment of classical phenylketonuria, Mol. Ther., 11, 986–989

Poppe, L. (2013) MIO-containing ammonia-lyases and 2,3-aminomutases. BioTrans. conference presentation, Manchester, UK

Prosekov, A. Y., Babich, O. O., Pokrovsky, V. S., Ivanova, A. S., Novoselova, V. M. (2014) Efficient expression of recombinant l-phenylalanine ammonia-lyase from Rhodosporidium toruloides using Escherichia coli, J. Appl. Biotechnol., 2, 2327-0640

Rétey, J. (2003) Discovery and role of methylidene imidazolone, a highly electrophilic prosthetic group, Biochim. Biophys. Acta, 1647, 179-184

Turner, N. (2011) Ammonia lyases and aminomutases as biocatalyst for the synthesis of α-amino and β-amino acids, Curr. Opin. Chem. Biol., 15, 234-240

STUDIA UBB BIOLOGIA, Volume 60 (LX), Sp.Issue, December 2015

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Published

2015-12-31

How to Cite

FILIP, A., BENCZE, L. C., PAIZS, C., POPPE, L., & IRIMIE, F. D. (2015). MIO-enzyme toolbox: cloning, expression and purification of recombinant „Rt”PAL. Studia Universitatis Babeș-Bolyai Biologia, 60(Sp.Issue), 39–43. Retrieved from http://193.231.18.162/index.php/subbbiologia/article/view/4625

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Volume 60, Sp.Issue, December 2015

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Articles

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