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Applied Microbiology and Biotechnology

, Volume 103, Issue 13, pp 5161–5166 | Cite as

Interferences that impact measuring optimal l-asparaginase activity and consequent errors interpreting these data

  • Marcela Medeiros de Freitas
  • Paula Monteiro Souza
  • Kellen Cruvinel
  • Thais Barros
  • Suikinai Nobre Santos
  • Paul F. Long
  • Adalberto Pessoa
  • Pérola Oliveira MagalhãesEmail author
Mini-Review
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Abstract

l-asparaginase is an enzyme produced by microorganisms, plants, and animals, which is used clinically for the treatment for acute lymphoblastic leukemia (ALL) and, in the food industry, to control acrylamide formation in baked foods. The purpose of this review was to evaluate the available literature regarding microbial sources of l-asparaginase, culture media used to achieve maximum enzyme expression in microbial fermentations, and assay methods employed to assess l-asparaginase activity. Studies were gathered by searching PubMed, and Web of Science databases before January 22, 2018, with no time restrictions. The articles were evaluated according to the source of l-asparaginase being studied, the nitrogen source in the culture medium, the type of sample, and the method employed to evaluate l-asparaginase activity. Bacterial l-asparaginase appeared to be the most commonly studied source of the enzyme and, most often, the enzyme activity was assayed from crude protein extracts using the Nessler method, which is an indirect measurement of asparaginase activity that determines the concentration of ammonia generated after the action of the enzyme on the substrate, l-asparagine. However, ammonia is also generated throughout microbial fermentations and this endogenous ammonia will also reduce the Nessler reagent if crude microbial extracts are used to determine total l-asparaginase activity. We suggest that current estimates of l-asparaginase activity reported in the literature may be overestimated when Nessler reagent is used, since we were unable to find a single study that made reference to the possible inference of fermentation derived ammonia.

Keywords

l-Asparaginase Enzyme assay Nessler method Interferences 

Notes

Acknowledgements

The authors are grateful to National Counsel of Technological and Scientific Development (CNPq), Support Research of the Federal District Foundation (FAPDF), and Higher Education Personnel Improvement Coordination (CAPES).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

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

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Marcela Medeiros de Freitas
    • 1
  • Paula Monteiro Souza
    • 1
  • Kellen Cruvinel
    • 1
  • Thais Barros
    • 1
  • Suikinai Nobre Santos
    • 2
  • Paul F. Long
    • 3
    • 4
  • Adalberto Pessoa
    • 4
  • Pérola Oliveira Magalhães
    • 1
    Email author
  1. 1.Natural Products Laboratory, Health Sciences SchoolUniversity of BrasíliaBrasíliaBrazil
  2. 2.Embrapa EnvironmentalJaguariúnaBrazil
  3. 3.School of Cancer & Pharmaceutical SciencesKing’s College LondonLondonUK
  4. 4.Department of Biochemical and Pharmaceutical TechnologyUniversity of São PauloSão PauloBrazil

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