L-Asparaginase is a therapeutic enzyme widely used as an antitumor agent for the treatment of acute lymphoblastic leukemia and lymphosarcoma. Its potential applications as a therapeutic agent and food additive require production at high yields in a low-cost medium with high purity and stability. Statistical experimental designs were applied to identify and optimize the significant parameters for the fermentative production of L-asparaginase by Pectobacterium carotovorum Microbial Type Culture Collection (MTCC) 1428. Screening of different carbon sources identified galactose as the best carbon source for the production of L-asparaginase. Twelve medium components were selected and sixteen experiments were carried out based on the Plackett-Burman (PB) design for identifying the most significant medium components. Yeast extract, galactose, monosodium glutamate, MgSO 4 .7H 2 O and CaCl 2 .2H 2 O were found to be the most significant components that affected the production of L-asparaginase, and they were optimized by response surface methodology (RSM) using central composite design. This optimized medium showed maximum production rates of periplasmic and extracellular L-asparaginase of 3.25 and 0.83 U ml -1 , respectively. The periplasmic L-asparaginase production in the optimized medium was twofold higher than in the basal medium. Unstructured kinetic models were applied and satisfactorily described the cell growth, product formation and substrate utilization profile, thus providing a better understanding and prediction of the fermentation profile. This paper details the application of PB experimental design and RSM for the optimization of L-asparaginase production from Pe. carotovorum MTCC 1428 in submerged fermentation.
- Pectobacterium carotovorum
- Plackett-Burman design
- Response surface methodology
- Unstructured kinetic modeling