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Enhanced production of glutaminase free L-asparaginase II by Bacillus subtilis WB800N through media optimization

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Abstract

We studied the crucial components which elevate the expression of recombinant novel glutaminase free L-asparaginase II (rL-asp II) from Bacillus subtilis WB800N. The Plackett-Burman tool identified sucrose, NH4Cl, NaH2-PO4 and MgSO4 as the significant influencing factors (p<0.05). Further investigations showed that artificial neural network-genetic algorithm (ANN-GA) was more effective than central composite design (CCD) in optimizing the influencing factors. The maximum rL-asp II expression was found to be 389.56 IU/ml and 525.98 IU/ml using CCD (R2=90.4%) and ANN-GA (R2=96.2%), respectively. The validation experiments were carried out in a 3 L batch bioreactor where kinetic modelling of the obtained data was done. The rL-asp II expressed effectively inhibiting the polyacrylamide formation in vitro where no solidification was observed, when 2ml of purified rL-asp II used even after 60 min of incubation. This is the first study to report highest production of rL-asp II in B. subtilis WB800N (525.98 IU/ml) till date by combining statistical designs with consecutive intermittent addition of IPTG in batch reactor.

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  1. Biochemical Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT) Guwahati, Guwahati, 781039, Assam, India

    Chityala Sushma, Ashish Prabhu Anand & Venkata Dasu Veeranki

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  1. Chityala Sushma
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Sushma, C., Anand, A.P. & Veeranki, V.D. Enhanced production of glutaminase free L-asparaginase II by Bacillus subtilis WB800N through media optimization. Korean J. Chem. Eng. 34, 2901–2915 (2017). https://doi.org/10.1007/s11814-017-0211-1

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