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Volume 69, Issue 12


Optimization and purification of anticancer enzyme L-glutaminase from Alcaligenes faecalis KLU102

Sureshbabu Pandian / Venkataraman Deepak
  • Department of Biotechnology, Kalasalingam University, Krishnankoil, 626126, Tamilnadu, India
  • School of Science and Technology, Nottingham Trent University, Nottingham, NG11 8NS, UK
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/ Shiva Sivasubramaniam
  • Department of Biotechnology, Kalasalingam University, Krishnankoil, 626126, Tamilnadu, India
  • School of Science and Technology, Nottingham Trent University, Nottingham, NG11 8NS, UK
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/ Hariharan Nellaiah / Krishnan Sundar
Published Online: 2015-01-06 | DOI: https://doi.org/10.2478/s11756-014-0486-1


L-Glutaminase, an amidohydrolase, is gaining importance on account of its potential anticancer activity. L-Glutaminase produced by Alcaligenes faecalis KLU102, isolated from the marine realm (Bay of Bengal), exhibits potential anticancer activity. Response surface methodology was employed for optimizing the medium composition. The concentrations of the various constituents were as follows: arabinose (2%), skim milk (4%), and salts viz. K2HPO4, KH2PO4, MgSO4, and NaCl (2%). The bacterium grown in the optimized medium yielded an enzyme activity of 1.34 ± 0.07 IU/mg in shake-flask cultures and this doubled (2.77 ± 0.35 IU/mg) when scale-up studies were conducted using a 3-L fermenter. The enzyme was purified to homogeneity using ion-exchange chromatography, and the purified enzyme was found to have a specific activity of 54.72 IU/mg, with a molecular weight of 37 kDa. Immobilization of the enzyme on PEG-PHB nanoparticles improved the stability of the enzyme significantly. Purified L-glutaminase exhibited cytotoxic activity against HeLa cells, as assayed by the MTT assay, with an IC50 value of 12.5 μg/mL.

Keywords: L-glutaminase; response surface methodology; PEG-PHB nanoparticle; anticancer

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About the article

Published Online: 2015-01-06

Published in Print: 2014-12-01

Citation Information: Biologia, Volume 69, Issue 12, Pages 1644–1651, ISSN (Online) 1336-9563, ISSN (Print) 0006-3088, DOI: https://doi.org/10.2478/s11756-014-0486-1.

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© 2014 Slovak Academy of Sciences. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

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