Batch, Fed Batch Production and Characterization of Glutaminase Free L-Asparaginase II of Pectobacterium carotovorum MTCC 1428 in Escherichia coli

DOI: 10.4236/aim.2014.410072   PDF   HTML     4,271 Downloads   5,169 Views   Citations


The present study describes the production optimization of recombinant L-asparaginase II of Pectobacterium carotovorum MTCC 1428 in Escherichia coli BL21 (DE3) at batch and fed batch bioreactor level. Production of recombinant L-asparaginase II in batch and fed batch mode was found to be 1.34 and 5.38 folds higher, respectively as compared to shake flask culture. SDS-PAGE and native PAGE of the purified enzyme revealed that molecular mass of the subunits and native enzyme are ~37.5 kDa and ~150 kDa, respectively. Optimum range of pH and temperature for hydrolysis of L-asparagine were found to be 7.5 - 8.5 and 47°C - 52°C, respectively. The recombinant enzyme is very specific for its natural substrate, L-asparagine. The activity of recombinant L-asparaginase II is improved by mono cations and diverse effectors including Na+, K+, L-cystine, L-histidine, glutathione and 2-mercaptoethanol whereas, it is moderately inhibited by different divalent cations and thiol group blocking reagent. The kinetic parameters Km, Vmax, kcat and Km/Kcat of purified recombinant L-asparaginase II were determined. The purified L-asparaginase II possesses no partial glutaminase activity, which is prerequisite to reduce the possibility of side effects during the course of anti-cancer therapy.

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Goswami, R. , Hegde, K. and Veeranki, V. (2014) Batch, Fed Batch Production and Characterization of Glutaminase Free L-Asparaginase II of Pectobacterium carotovorum MTCC 1428 in Escherichia coli. Advances in Microbiology, 4, 667-680. doi: 10.4236/aim.2014.410072.

Conflicts of Interest

The authors declare no conflicts of interest.


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