Functional parametric influence on glutaminase free l-asparaginase production by Amycolatopsis thermoflava SFMA-103

l-Asparaginase (l-ASNase) is a well known chemotherapeutic enzyme, to treat different neoplasms, especially in treating Acute Lymphoblastic Leukaemia (ALL) of children and adults. However, production of glutaminase free l-ASNase is the central area of research to produce cytotoxicity free L-ASNases in the cancer therapeutics. The present study explored the potential glutaminase free l-ASNase production from rare actinobacterium, Amycolatopsis thermoflava SFMA-103 isolated from agricultural field soils. The attention of the enzyme production was sequentially optimized using various methods starting from One-factor-at-a-time (OFAT) to Taguchi orthogonal array (OA) experimental design (DOE). Four critical factors, pH, temperature, arabinose and yeast extract were identified initially, hence, three levels with a layout of L9 (34) were used and further processed with Qualitek-4 software. Upon validation an enhanced enzyme production of 68.55% was observed (from 10 IU −1 min−1 to 31.8 IU −1 min−1). As far as our knowledge extends, glutaminase-free l-ASNase production is the first time report from Amycolatopsis thermoflava SFMA-103. Further, scale-up studies were performed using bench scale bioreactor (5L capacity) with the statistically optimized conditions (aeration of 0.5 VVM and agitation of 150 rpm at 40 °C) resulted in an improved enzyme production to the level of 132.69% (61.6 IU −1 min−1) with an incubation period of 66 h. Taguchi methods for scale-up studies of l-ASNase proves useful in bioprocess optimization with minor experimental set up, saving cost and time consumption is low. [Display omitted] •Isolated Amycolatopsis thermoflava SFMA-103, a rare actinobacterium having potential to produce glutaminase free l-Asparaginase (ASNase).•ASNase production optimized using Taguchi orthogonal array experimental design.•pH, temperature, arabinose and yeast extract were noticed as significant factors and at optimized condition production yield was improved >68%.•Bioreactor based scale up studies enhanced the enzyme production to the tune of 6 fold upon step up optimization.