Abstract
With an increasing demand for l-asparaginase in pharmaceutical and food sectors for its cytostatic and acrylamide-reducing qualities, there's a need to discover novel, highly productive enzyme sources with improved pharmacokinetic profiles. Keeping this in mind, the present study aimed at maximizing the potential of Ganoderma australe GPC191 to produce l-asparaginase by fermentation medium optimization using statistical validation. Of the 11 physicochemical parameters evaluated under submerged fermentation conditions through one-factor-at-a-time approach and Plackett–Burman design, only four parameters (inoculum load, l-asparagine, soybean meal, and initial pH) influenced l-asparaginase production, significantly (p < 0.001). The optimal levels and interaction effects of these on the overall production were further evaluated by the central composite rotatable design of response surface methodology. Post-optimization, 27.34 U/mL was predicted as the maximum activity at pH 7 with 5n inoculum load and 15 g/L each of l-asparagine and soybean meal. Experimental validation yielded an activity of 28.52 U/mL, indicating an overall 18.17-fold increase from the unoptimized stage. To our knowledge, this is the first report signifying the l-asparaginase production aptitude of G. australe with sequential statistical validation using agricultural waste, which can serve as a model to enhance its yields, offering a sustainable and cost-effective solution for industrial application.
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Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Code availability
GraphPad Prism software (version 9.2.0); Design Expert software (13.0 version).
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Acknowledgements
We sincerely thank the management of JAIN (Deemed-to-be University) for providing research facilities. The authors are grateful to Dr. Sourav Bhattacharya, Assistant Professor, Department of Microbiology and Botany, JAIN (Deemed-to-be University), for supporting the execution of OFAT experiments. We also thank Dr. Sushil Kumar Bhattacharyya for his guidance in proofreading the article. All authors concur with the submission. The work has not been published elsewhere, either completely, in part, or in another form. The manuscript has not been submitted to another journal and will not be published elsewhere. The manuscript does not contain experiments using animals. The manuscript does not contain human studies.
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SS and MC contributed to the concept development and design for the research. MC executed, analyzed, and interpreted the study under the guidance of SS. MC developed the manuscript draft, which was reviewed and edited by SS. All authors have seen and approved the final manuscript and its contents and are aware of the responsibilities connected to the authorship.
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Chakraborty, M., Shivakumar, S. Application of sequential design for enhanced l-asparaginase synthesis from Ganoderma australe GPC191. World J Microbiol Biotechnol 40, 85 (2024). https://doi.org/10.1007/s11274-023-03881-0
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DOI: https://doi.org/10.1007/s11274-023-03881-0