Validation of a Simple and Rapid Method for Assessment of Intracellular Bacterial Asparaginase Validation of a Simple Method for Assessment of Bacterial Asparaginase
Iranian Journal of Pharmaceutical Sciences,
Vol. 12 No. 2 (2016),
1 April 2016
,
Page 33-42
https://doi.org/10.22037/ijps.v12.40776
Abstract
L-Asparaginase has remarkable properties which make it useful in dual pharmaceutical and food industries.In this study, simple and advantageous methods have been validated for rapid and precise
determination of intracellular L-Asparaginasein bacterial species. A suspension of bacterial cells was used instead of cell extract and incubated by substrate (asparagine) after simple wash and centrifugation steps. Due to loss of enzyme activity which could be caused by cell distruption methods such as sonication or enzymatic treatment, cell suspension was used instead of the cell extract. Thus, this method not only is cost effective but also speeds up the screening process and leasd to higher measurement accuracy. To validate this method, two species of bacteria; E.coli ATCC 8739 and Halomonas H28 were used. After cultivation, the cells were harvested and washed. Then, 5 serial dilutions were prepared from each bacterium, and the asparaginase activity in each of them was measured by methods including sonication, enzymatic lyses, and the cell suspension. The results have showed that the changes in asparaginase activity in all 5 serial dilutions are linear and there is good agreement between the sonication and the cell suspension methods. Also, it was shown that activities measured by the enzymatic method were significantly higher than the other two methods.
- Enzymatic lyses
- Intracellular
- L-Asparaginase
- Sonication
- E. Coli
- Halomonas
How to Cite
References
[2] Siddalingeshwara K and Lingappa K. Screening and optimization of L-asparaginase-A tumour inhibitor from Aspergillus terreus through solid state fermentation. J Adv Sci Res (2010)1(1):55-60.
[3] Müller H and Boos J. Use of L-asparaginase in childhood ALL. Crit Rev Oncol Hematol (1998) 28(2): 97-113.
[4] Kukurová K, et al. Raising agents strongly influence acrylamide and HMF formation in cookies and conditions for asparaginase activity in dough. Eur Food Res Technol (2013) 237(1): 1-8.
[5] Hendriksen HV, et al. Evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from Aspergillus oryzae. J Agric Food Chem (2009) 57(10): 4168-4176.
[6] Boegl ZC. Impact of L-asparaginase on acrylamide content in potato products. J Food Nutr Res (2006) 45(4): 141-146.
[7] Pedreschi FK, aack K and Granby K. The effect of asparaginase on acrylamide formation in French fries.Food Chem (2008) 109(2): 386-392.
[8] Mottram DS, Wedzicha BL and Dodson AT. Food chemistry: acrylamide is formed in the Maillard reaction. Nature (2002) 419(6906): 448-449.
[9] Duval M, et al. Comparison of Escherichia coli–asparaginase withErwinia-asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research
and Treatment of Cancer—Children's Leukemia Group phase 3 trial. Blood (2002) 99(8): 2734-2739.
[10] Warangkar SC and Khobragade CN. Purification, characterization, and effect of thiol compounds on activity of the Erwinia carotovora L-asparaginase.Enzyme Res(2009) 1:2010.
[11] Wriston J and Yellin T. L-asparaginase: a review. Adv Enzymol Relat Areas Mol Biol (1973) 39:185-248.
[12] Savitri AN and Azmi W. Microbial Lasparaginase:A potent antitumour enzyme. Indian J Biotechnol (2003) 2: 184-194.
[13] Khamna S, Yokota A and Lumyong S. LAsparaginase production by actinomycetes isolated from some Thai medicinal plant rhizosphere soils. Int j integr biol (2009) 6(1): 22-26.
[14] Barati, M., et al., L-Asparaginase Activity in Cell Lysates and Culture Media of Halophilic Bacterial Isolates. Iranian Journal of Pharmaceutical Research, 2016. 15(3): p. 435-440.
[15] Imada, A., et al., Asparaginase and glutaminase activities of micro-organisms. Microbiology, 1973.76(1): p. 85-99.
[16] Passing, H. and W. Bablok, A new biometrical procedure for testing the equality of measurements from two different analytical methods. Application of linear regression procedures for method comparison
studies in clinical chemistry, Part I. Clinical Chemistry and Laboratory Medicine, 1983. 21(11): p.709-720.
[17] Altman, D.G. and J.M. Bland, Measurement in medicine: the analysis of method comparison studies.The statistician, 1983: p. 307-317.
- Abstract Viewed: 288 times
- IJPS_Volume 12_Issue 2_Pages 33-42 Downloaded: 174 times