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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39 For more details click here
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2019: 96.39
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2020.9(3): 260-287
DOI: https://doi.org/10.20546/ijcmas.2020.903.032


Optimization of Extraction Techniques for the Release of Intracellular L-Asparginase from Serratia marcescens MTCC 97 and its Characterization
Manisha Gautam*, Nisha and Wamik Azmi
Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla (H.P.) 171005, India
*Corresponding author
Abstract:

L-asparaginase acts as an efficient agent in curing certain sorts of lymphoma and leukemia by catalyzing the deamination of L-asparagine to L-aspartate and ammonia. Microorganisms are better source of L-asparginase, as their culturing, extraction and purification is more convenient than plants and other sources. As most of L-asparginases are intracellular in nature, so the selection of a suitable method for its release with maximum recovery was become more important. In present study, the resting cells of S. marcescens MTCC 97 were disintegrated by different enzymatic (lysozyme), chemical (alkali lysis, acetone powder, guanidine-HCl and triton X-100) and physical (motor and pestle, vortex, bead beater and sonicator) methods. Among all methods explored, sonication was found best method with 0.05 U/mg specific activity and minimum loss of enzyme (8%). Different reaction parameters were also optimized for the characterization of released L-asparginase. The extracted L-asparaginase showed maximum activity (0.985 U/ml) in 0.05M sodium phosphate buffer (pH 7.5) with L-asparagine (8mM) as substrate at 40oC incubation for 20 min. Moreover, different metal ions, additives, chelating agents and protease inhibitors showed negative effects on L-asparaginase activity of resting cells and cell free extract obtained from S. marcescens MTCC 97.


Keywords: L-asparaginase, Serratia marcescens MTCC 97, disintegration, sonication.
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How to cite this article:

Manisha Gautam, Nisha and Wamik Azmi. 2020. Optimization of Extraction Techniques for the Release of Intracellular L-Asparginase from Serratia Marcescens MTCC 97 and its Characterization.Int.J.Curr.Microbiol.App.Sci. 9(3): 260-287. doi: https://doi.org/10.20546/ijcmas.2020.903.032