Recombinant l-Asparaginase II from Lactobacillus casei subsp. casei ATCC 393 and Its Anticancer Activity

Indian Journal of Microbiology
S Susan AishwaryaKandathil Narayanan Rajnish

Abstract

l-asparaginases from bacterial origin are employed extensively in leukemic treatment and food industry. The present study focuses on the characterization of the recombinant l-asparaginase II from Lactobacillus casei subsp. casei ATCC 393 cloned into Escherichia coli expression system and purified using Ni-NTA chromatography. The recombinant l-asparaginase as a monomer had a molecular weight of 35 kDa. The enzyme was active from 10 to 80 °C with the optimum at 40 °C. The enzyme retained its activity at 28 °C and 37 °C up to 24 h. The enzyme had optimum pH of 6 and retained 50% activity till 18 h. The Km of the recombinant enzyme was 0.01235 mM and Vmax 1.576 mM/min. The half life of recombinant l-asparaginase II in human serum was 44 h and trypsin was for 15 min. The LC-MS/MS analysis revealed the molecular weight of 35,050 and pI of 5.64. The secondary structure prediction using CD spectroscopy for the recombinant enzyme showed 33.5% α-helix, 66.5% turn and 0% β sheets. The cytotoxicity of the recombinant enzyme was analysed against MOLT 3, Jurkat E6.1 and K-562 with the IC 50 value of 30, 62.5 and 50 µg/ml.

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Citations

Oct 23, 2020·Indian Journal of Microbiology·Susan Aishwarya SureshK N Rajnish
May 1, 2021·International Journal of Environmental Research and Public Health·Cristina SarionAdriana Dabija
Jun 2, 2021·Applied Microbiology and Biotechnology·Daniel CastroValéria C Santos-Ebinuma
Jul 9, 2021·Applied Microbiology and Biotechnology·Nidhi Y PatelVikram H Raval
Sep 18, 2021·Environmental Science and Pollution Research International·Sulaiman A Al Yousef

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Methods Mentioned

BETA
PCR
enzyme assay
enzyme
column chromatography

Software Mentioned

GraphPad Prism
VERIFY
ExPASy
ProtParam tool
PRALINE
CLUSTALW
Easy Modeller
Mascot
ProtParam
SignalP

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