Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Production of Extracellular Anti-leukaemic Enzyme L-asparaginase from Marine Actinomycetes by Solid-state and Submerged Fermentation: Purification and Characterisation

Saleem N Basha¹ , R Rekha², M Komala¹, S Ruby³

¹Department of Pharmaceutical Biotechnology; ²Department of Pharmacognosy; ³Department of Pharmaceutical Chemistry, Mohamed Sathak A J College of Pharmacy, Medavakkam, Chennai 600 119, India.

For correspondence:- Saleem Basha Email: nsaleem_basha@rediffmail.com Tel:+91449943998065

Received: 23 November Accepted: 16 May 2009 Published: 14 August 2009

Citation: Basha SN, Rekha R, Komala M, Ruby S. Production of Extracellular Anti-leukaemic Enzyme L-asparaginase from Marine Actinomycetes by Solid-state and Submerged Fermentation: Purification and Characterisation. Trop J Pharm Res 2009; 8(4):353-360 doi: 10.4314/tjpr.v8i4.10

© 2009 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: The objective of this investigation was to isolate marine actinomycetes, screen them for L-asparaginase activity and characterise the enzyme.

Methods: Marine actinomycetes were isolated from sediment samples obtained from Tamilnadu and Kerala in India. The isolates were identified as actinomycetes by microscopical and biochemical tests. Production of L-asparaginase was carried out in three different media, namely, solid-state media, Tryptone Glucose Yeast extract (TGY) broth and Tryptone Fructose Yeast extract (TFY) broth.. The enzyme was purified to near homogeneity by ammonium sulphate precipitation, dialysis, gel filtration on Sephadex G-100 column and SDS-PAGE.

Results: Among 10 marine isolates subjected to preliminary screening, only isolates S3, S4 and K8 showed potential for L-asparaginase activity. All three marine soil isolates synthesized asparaginase with yield ranging from 24.6 to 49.2 IU/ml. Soil isolate S3 showed the highest productivity of 49.2 IU/ml with a protein content of 65 µg/ml and optimum activity at pH 7.5 and 50 ºC. The apparent K_m value for the substrate was 25 µM. Mg²⁺ ion slightly stimulated activity while Cu²⁺, Zn²⁺ and EDTA were inhibitory.

Conclusion: The study revealed that marine actinomycetes may be a potential source of high yield, high substrate specificity L-asparaginase, which is an anti-leukaemia agent.

Keywords: L-asparaginase, Solid-state media, TGY broth, TFY broth, SDS-PAGE, Km and Vmax