title: Low cost single-step purification of endoglucanase from Aspergillus fumigatus ABK-9.
abstract: Low cost agro-waste was used as adsorption support for single-step purification of endoglucanase from the culture filtrate of A. fumigatus ABK-9. Among various agro-waste substrates, 1% NaOH pretreated rice bran was proved to be the best for adsorbing about 74.8 and 71.1% of endoglucanase at 4 degrees C and 10 degrees C respectively. Langmuir type adsorption isotherm at 4 degrees C showed maximum adsorption of enzyme at pH 5.0, which was in the range of optimum pH of the enzyme. The rice bran column bound enzyme was maximally eluted by a mixture of acetate buffer (0.05 M, pH 5.5) and ethanol (40%, v/v) at a ratio of 3:2 and a flow rate of 1 mL/min. A 5.52-fold purification of the enzyme was achieved from culture supernatant. The specific activity and recovery yield after purification were 294.0 U/mg and 40.15%, respectively, which were comparable with other contemporary protocols. The homogeneity of the enzyme was tested through sodium dodecyl sulphate polyacrylamide gel electrophoresis and a single band of 56.3 kDa was observed. Zymogram analysis finally confirmed the occurrence of endoglucanase in the single band.
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Arpan
authors.initials: A
authors.lastname: Das
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Tanmay
authors.initials: T
authors.lastname: Paul
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Suman K
authors.initials: SK
authors.lastname: Halder
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Arijit
authors.initials: A
authors.lastname: Jana
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Kuntal
authors.initials: K
authors.lastname: Ghosh
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Chiranjit
authors.initials: C
authors.lastname: Maity
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Das Mohapatra
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Bikas R
authors.initials: BR
authors.lastname: Pati
authors.affiliation: Department of Microbiology, Vidyasagar University, Midnapore 721 102, India.
authors.forename: Keshab C
authors.initials: KC
authors.lastname: Mondal
mesh_terms: D000327:Adsorption; D001232:Aspergillus fumigatus / Q000201:enzymology*; D002480:Cellulase / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D013696:Temperature; D013816:Thermodynamics
chemical_list: D002480:Cellulase