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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abd El Baky HH, El Baroty GS (2016) Optimization of growth conditions for purification and production of L-asparaginase by Spirulina maxima. J Evid Based Complementary Altern Med: eCAM 2016:1785938. https://doi.org/10.1155/2016/1785938
Alhussaini MS (2013) Mycobiota of wheat flour and detection of alpha- amylase and L-asparaginase enzymes. Life Sci J-Acta Zhengzhou University Overseas Edition 10(1):1112–1122
Arrivukkarasan S, Muthusivaramapandian M, Aravindan R, Viruthagiri T (2010) Effect of medium composition and kinetic studies on extracellular and intracellular production of L-asparaginase from Pectobacterium carotovorum. Food Sci Technol Int 16(2):115–125. https://doi.org/10.1177/1082013209353219
Balcao VM, Mateo C, Fernandez-Lafuente R, Malcata FX, Guisan JM (2001) Coimmobilization of L-asparaginase and glutamate dehydrogenase onto highly activated supports. Enzyme Microb Technol 28(7-8):696–704
Cedar H, Schwartz JH (1967) Localization of the two-L-asparaginases in anaerobically grown Escherichia coli. J Biol Chem 242(16):3753–3755
Cedar H, Schwartz JH (1968) Production of L-asparaginase II by Escherichia coli. J Bacteriol 96(6):2043–2048
Chow Y, Ting AS (2015) Endophytic l-asparaginase-producing fungi from plants associated with anticancer properties. J Adv Res 6(6):869–876. https://doi.org/10.1016/j.jare.2014.07.005
Credali A, Garcia-Calderon M, Dam S, Perry J, Diaz-Quintana A, Parniske M, Wang TL, Stougaard J, Vega JM, Marquez AJ (2013) The K+-dependent asparaginase, NSE1, is crucial for plant growth and seed production in Lotus japonicus. Plant Cell Physiol 54(1):107–118. https://doi.org/10.1093/pcp/pcs156
Drainas D, Drainas C (1985) A conductimetric method for assaying asparaginase activity in Aspergillus nidulans. Eur J Biochem 151(3):591–593
Drainas C, Kinghorn JR, Pateman JA (1977) Aspartic hydroxamate resistance and asparaginase regulation in the fungus Aspergillus nidulans. J Gen Microbiol 98(2):493–501
Dunlop PC, Roon RJ (1975) L-Asparaginase of Saccharomyces cerevisiae: an extracellular Enzyme. J Bacteriol 122(3):1017–1024
Fry BA (1955) The Nitrogen Metabolism of Microorganisms. Methuen, London
Gulati R, Saxena RK, Gupta R (1997) A rapid plate assay for screening L-asparaginase producing microorganisms. Lett Appl Microbiol 24(1):23–26
Hernadi F, Polya K, Daroczy A (1971) Production of L-asparaginase isoenzymes by strains E. coli. Archiv fur Mikrobiol 77(2):151–154
Imada A, Igarasi S, Nakahama K, Isono M (1973) Asparaginase and glutaminase activities of microorganisms. J Gen Microbiol 76(1):85–99
Jayaram HN, Cooney DA, Jayaram S, Rosenblum L (1974) A simple and rapid method for the estimation of L-asparaginase in chromatographic and electrophoretic effluents: comparison with other methods. Anal Biochem 59(2):327–346
Lopes AM, Oliveira-Nascimento L, Ribeiro A, Tairum CA Jr, Breyer CA, Oliveira MA, Monteiro G, Souza-Motta CM, Magalhaes PO, Avendano JG, Cavaco-Paulo AM, Mazzola PG, Rangel-Yagui CO, Sette LD, Converti A, Pessoa A (2015) Therapeutic L-asparaginase: upstream, downstream and beyond. Crit Rev Biotechnol:1–18. https://doi.org/10.3109/07388551.2015.1120705
Magri A, Soler MF, Lopes AM, Cilli EM, Barner PS, Pessoa A, Pereira JFB (2018) A critical analysis of L-asparaginase activity quantification methods - colorimetric methods versus high-performance liquid chromatography. Anal Bioanal Chem 410:6985–6990. https://doi.org/10.1007/s00216-018-1326-x
Mahajan RV, Saran S, Saxena RK, Srivastava AK (2013) A rapid, efficient and sensitive plate assay for detection and screening of L-asparaginase-producing microorganisms. FEMS Microbiol Lett 341(2):122–126. https://doi.org/10.1111/1574-6968.12100
Pajdak E, Pajdak W (1972) A simple and sensitive method for detection of L-asparaginase by polyacrylamide gel electrophoresis. Anal Biochem 50(1):317–320
Pajdak E, Pajdak W (1974) Activity stain for the detection of L-asparaginase-anti-L-asparaginase complexes in agar. J Immunol Methods 6(1):109–114. https://doi.org/10.1016/0022-1759(74)90095-7
Pronk MEJ, Verger P, Olempska-Beer Z, Walker R (2008) Asparaginase from Aspergillus oryzae expressed in Aspergillus oryzae. WHO Food Additives Series 59:55–63
Roth G, Nunes JES, Rosado LA, Bizarro CV, Volpato G, Nunes CP, Renard G, Basso LA, Santos DS, Chies JM (2013) Recombinant Erwinia carotovora L-asparaginase II production in Escherichia coli fed-batch cultures. Braz J Chem Eng 30(2):245–256. https://doi.org/10.1590/s0104-66322013000200003
Russell JA (1944) The colorimetric estimation of small amounts of ammonia by the phenol-hypochlorite reaction. J Biol Chem 156:457–462
Souza PM, Freitas MM, Cardoso SL, Pessoa A, Guerra ENS, Magalhães PO (2017) Optimization and purification of L-asparaginase from fungi: a systematic review. Crit Rev Oncol Hematol 120:194–202. https://doi.org/10.1016/j.critrevonc.2017.11.006
Stein LY, Klotz MG (2016) The nitrogen cycle. Curr Biol 26(3):R94–R98. https://doi.org/10.1016/j.cub.2015.12.021
Tagami S, Matsuda K (1990) An enzymatic method for the kinetic measurement of L-asparaginase activity and L-asparagine with an ammonia gas-sensing electrode. Chem Pharm Bull 38(1):153–155
Teixeira CSS, Fernandes HS, Fernandes PA, Ramos MJ, Cerqueira NMFSA (2017) Cancer therapies based on enzymatic amino acid depletion. In: Nanostructures for Cancer Therapy. Elsevier, pp 623–651
Thenmozhi C, Sankar R, Karuppiah V, Sampathkumar P (2011) L-asparaginase production by mangrove derived Bacillus cereus MAB5: optimization by response surface methodology. Asian Pac J Trop Med 4(6):486–491. https://doi.org/10.1016/s1995-7645(11)60132-6
Whitecar JPJ, Bodey GP, Harris JE, Freireich EJ (1970) L-Asparaginase. N Engl J Med 282(13):732–734. https://doi.org/10.1056/nejm197003262821307
Ylikangas P, Mononen I (2000) A fluorometric assay for L-asparaginase activity and monitoring of L-asparaginase therapy. Anal Biochem 280(1):42–45. https://doi.org/10.1006/abio.2000.4500
Zhou L, Boyd CE (2016) Comparison of Nessler, phenate, salicylate and ion selective electrode procedures for determination of total ammonia nitrogen in aquaculture. Aquacult 450:187–193. https://doi.org/10.1016/j.aquaculture.2015.07.022
Zuo S, Zhang T, Jiang B, Mu W (2015) Recent research progress on microbial L-asparaginases. Appl Microbiol Biotechnol 99(3):1069–1079. https://doi.org/10.1007/s00253-014-6271-9
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).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
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.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
de Freitas, M.M., Souza, P.M., Cruvinel, K. et al. Interferences that impact measuring optimal l-asparaginase activity and consequent errors interpreting these data. Appl Microbiol Biotechnol 103, 5161–5166 (2019). https://doi.org/10.1007/s00253-019-09890-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-019-09890-0