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l-Asparaginase production in rotating bed reactor from Rhizopus microsporus IBBL-2 using immobilized Ca-alginate beads

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The present work reports on the production of extracellular l-asparaginase from Rhizopus microsporus IBBL-2 using submerged fermentation (SmF) process free of glutaminase and urease activities. Primary studies done in shake flask showed that the highest l-asparaginase activity of 12.68 U mL−1 was produced at 72 h with optimized fermentation parameters such as pH 6.0, 4 × 106 fungal cells mL−1, and agitation of 180 rpm at 30 °C using one-factor-at-a-time (OFAT). Different substrates, nitrogen sources, temperature, pH, the initial number of cells and metal ions were tested to determine the impact on enzyme production. l-Asparaginase activity of 17.68 U mL−1 was produced after 48 h using immobilized calcium-alginate (Ca-alginate) cells. 4 × 106 cells mL−1 was entrapped in 3% (W/V) of alginate bead of size 2 mm each at a temperature of 30 °C and pH of 6. The process was optimized using L9 (34) Taguchi Orthogonal Array (OA) technique with a regression coefficient (R2) value of 0.9709, F value of 33.34 and p value of 0.0025. Scale-up studies involving 200-mL and 1-L rotating bed reactor (RBR) using immobilized beads were done and the results obtained are 20.21 U mL−1 and 19.13 U mL−1, respectively, the increased activity with immobilization accounts for reduced shear on cells due to increased stability as compared to the free-flowing cells.

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This work is supported by a research grant from the Department of Science and Technology-Science and Engineering Research Board SERB (SB-EMEQ-048/2014). The authors would like to express their sincere gratitude to Dr. Kruthi Doriya, Vaibhav Lendekar, Venkatesh Mandari, and Ashutosh Nema for their support during the experimentation process.

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Anup Ashok has done the primary work including experimentation, analysis and writing the paper. Dr. Devarai Santhosh Kumar has supervised the entire process and supported in the experimentation, analyses and correction of the manuscript.

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Correspondence to Santhosh Kumar Devarai.

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On behalf of all authors, the corresponding author states that there is no conflict of interest.

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Ashok, A., Devarai, S.K. l-Asparaginase production in rotating bed reactor from Rhizopus microsporus IBBL-2 using immobilized Ca-alginate beads. 3 Biotech 9, 349 (2019).

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