Optimization of culture variables for the production of L- asparaginase from Pectobacterium carotovorum

Abstract

L-Asparaginase is an anti-neoplastic agent used in the chemotherapy of acute lymphoblastic leukemia. The present work deals with production of intra-cellular L-asparaginase from Pectobacterium carotovorum using submerged fermentation. The statistical approaches of Plackett-Burman and response surface methodology were applied to optimize the culture parameters for the production of L-asparaginase through a bacterium P. carotovorum MTCC 1428. Optimal conditions for maximal productivity were determined based on six parameters at two different levels. Initially, a screening design methodology was used to evaluate the process variables, and the response surfaces applied to find optimal concentration for production. The analysis revealed that the optimum levels of L-asparagine, lactose and inoculum percentage were found to be 4.00, 3.5 g L⁻¹, and 2.75%, respectively. The maximum enzyme activity in the optimized medium was 19.33 U ml⁻¹.

Keywords: Design of experiments (DoE), L-asparaginase, response surface methodology, Plackett-Burman design and acute lymphoblastic leukemi.

Afr. J. Biotechnol Vol. 12 No. 50