title: Optimization, purification, and characterization of L-asparaginase from Actinomycetales bacterium BkSoiiA.
abstract: Actinobacteria are promising source of a wide range of important enzymes, some of which are produced in industrial scale, with others yet to be harnessed. L-Asparaginase is used as an antineoplastic agent. The present work deals with the production and optimization of L-asparaginase from Actinomycetales bacterium BkSoiiA using submerged fermentation in M9 medium. Production optimization resulted in a modified M9 medium with yeast extract and fructose as carbon and nitrogen sources, respectively, at pH 8.0, incubated for 120 hr at 30 ± 2 °C. The crude enzyme was purified to near homogeneity by ammonium sulfate precipitation following dialysis, ion-exchange column chromatography, and finally gel filtration. The sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) revealed an apparent molecular weight of 57 kD. The enzyme was purified 95.06-fold and showed a final specific activity of 204.37 U/mg with 3.49% yield. The purified enzyme showed maximum activity at a pH 10.0 and was stable at pH 7.0 to 9.0. The enzyme was activated by Mn(2+) and strongly inhibited by Ba(2+). All these preliminary characterization suggests that the L-asparaginase from the source may be a tool useful to pharmaceutical industries after further research.
authors.affiliation: a Department of Microbiology , Orissa University of Agriculture and Technology , Bhubaneshwar , Odisha , India.
authors.forename: Chitrangada
authors.initials: C
authors.lastname: Dash
authors.affiliation: a Department of Microbiology , Orissa University of Agriculture and Technology , Bhubaneshwar , Odisha , India.
authors.forename: Sukanti Bala
authors.initials: SB
authors.lastname: Mohapatra
authors.affiliation: b Imgenex India Pvt. Ltd. , Bhubaneshwar , Odisha , India.
authors.forename: Prasanta Kumar
authors.initials: PK
authors.lastname: Maiti
mesh_terms: D000192:Actinomycetales / Q000201:enzymology*; D001215:Asparaginase / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D013696:Temperature
chemical_list: D001215:Asparaginase
keywords: Actinomycetales bacterium BkSoiiA; L-asparaginase; enzyme; optimization; purification; submerged fermentation