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Response surface methodology based optimized production, purification, and characterization of L-asparaginase from Fusarium foetens

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Abstract

L-asparaginase is used as one of the prime chemotherapeutic agents to treat acute lymphoblastic leukemia. L-asparaginase obtained from bacteria exhibits hypersensitive reactions including various side effects. The present work aimed to optimize growth parameters for maximum production of L-asparaginase by Fusarium foetens through response surface methodology, its purification, and characterization. The optimization of L-asparaginase production by Fusarium foetens was initially done through a one-factor-at-a-time method. L-asparaginase production was further optimized using a central composite design based response surface methodology. The maximum L-asparaginase activity of 12.83 IU/ml was obtained under the following growth conditions; temperature-27.5 °C, pH-8, inoculum concentration-1.5 × 106 spores/ml, and incubation period-7 days. In comparison with the unoptimized growth conditions (4.58 IU/ml), the optimization led to a 2.65-fold increase in the L-asparaginase activity. The L-asparaginase from Fusarium foetens was purified 15.60-fold, with a yield of 39.89% using DEAE-cellulose column chromatography. After purification, the L-asparaginase activity was determined to be 127.26 IU/ml and the specific activity was found to be 231.38 IU/mg. The molecular mass was estimated to be approximately 37 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified enzyme showed optimum activity at pH 5, and a temperature of 40 °C. The enzyme showed 100% specificity towards L-asparagine and no activity towards L-glutamine. Its activity was enhanced by Mn2+, Fe2+, and Mg2, while it was inhibited by β-mercaptoethanol and EDTA. The Km and Vmax of the purified L-asparaginase were found to be 23.82 mM and 210.3 IU/ml respectively. The results suggest that Fusarium foetens could be a potent candidate for the bioprocessing of L-asparaginase at a large scale.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors are thankful to the Council of Scientific and Industrial Research (CSIR) for providing a Junior Research Fellowship (JRF) and DOS in Microbiology, University of Mysore for providing facilities.

Funding

The study was funded by CSIR (Council of Scientific and Industrial Research) under Junior Research Fellowship (File No:09/119(0218)/2019-EMR-I).

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Authors and Affiliations

  1. Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysuru, Karnataka, 570 006, India

    Javaraiah Parashiva, Bettadapura Rameshgowda Nuthan, Purushotham Tejashwini & Sreedharamurthy Satish

  2. Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysuru, Karnataka, 570 005, India

    Madeva Bharatha & Raju Praveen

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  1. Javaraiah Parashiva
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All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Javaraiah Parashiva. The first draft of the manuscript was written by Javaraiah Parashiva and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Parashiva, J., Nuthan, B.R., Bharatha, M. et al. Response surface methodology based optimized production, purification, and characterization of L-asparaginase from Fusarium foetens. World J Microbiol Biotechnol 39, 252 (2023). https://doi.org/10.1007/s11274-023-03684-3

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