Production, purification and characterization of L-asparaginase from Streptomyces gulbargensis

Abstract

L-asparaginase is an anti-neoplastic agent used in the lymphoblastic leukaemia chemotherapy. In the present study a novel strain, Streptomyces gulbargensis was explored for the production of extra-cellular L-asparaginase using groundnut cake extract. The optimum pH, temperature, inoculum size and agitation speed for enzyme production were pH 8.5, 40°C, 1x10 8 spores/ml and 200 rev/min respectively. Maltose (0.5%) and L-asparagine (0.5%) proved to be the best carbon and nitrogen sources respectively. The enzyme was purified 82.12 fold and the apparent molecular weight of the enzyme was found to be 85 kDa.The optima pH and temperature for the enzyme were 9.0 and 40 o C respectively. The enzyme was more stable at the alkaline pH than at the acidic one and it retained 55% of the activity at 80 o C for 60 min.

Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

“…BCCS 034 (1.64 IU/mL) in the supernatant as reported by Kumar and Ebrahiminezhad with their associates [9,16] .…”

“…LAse produced extracellularly is advantageous and preferred over intracellular type because of higher accumulation of protein, easy extraction and downstream processing [16] . The extracellular compartment in bacteria is protease deficient and the liberated protein exported to the medium is mostly soluble, biologically active and has an authentic N-terminus, relatively free from endotoxins those results in minimization of adverse effects.…”

“…The extremozymes or thermozymes from thermophilic organisms are preferred owing to their guaranteed properties of stability in adverse conditions and different industrial process at which they operate [20] . produced LAse extracellularly, that is advantageous and preferred over intracellular type because of higher accumulation of mostly soluble biologically active protein, easy extraction, downstream processing [16] and relatively free from endotoxins those results in minimization of adverse effects. The isolate TPS-12 is growing in the temperature range of 150-70°C, which confirm its thermophilic characteristic.…”

Comparative Studies on Effect of Carbon and Nitrogen Sources on L -Asparaginase Production

“…These enzymes are selectively killed by L-asparagine deprivation. The clinical action of this enzyme is attributed to the reduction of L-asparagine (Amena et al, 2010). Lasparaginase production by microorganisms are basically used in clinical practices because the diversity of microbial population is being explored for gathering novel sources of L-asparaginase production with less side effects and better treatment (Geckil and Gencer, 2004).…”

Purification and Cytotoxicity of L-asparaginase from Streptomyces griseoluteus GDJ1

“…The Sephadex G-100 and Sephadex C-50 chromatography was found as important for the separation and purification of L-asparaginase. By this purification procedure, the strain Streptomyces griseoluteus GDJ1 (AT-R-S1) has produced L-asparaginase with 119 fold purity which is far better than compared to Streptomyces gulbargensis which has shown only 82.12 fold purity [28]. (5µg -45µg/ml) for 72 hours and the results were shown in Figure 21.…”