| Autor: |
Arpan, Das, Tanmay, Paul, Suman K, Halder, Arijit, Jana, Kuntal, Ghosh, Chiranjit, Maity, Pradeep K, Das Mohapatra, Bikas R, Pati, Keshab C, Mondal |
| Rok vydání: |
2014 |
| Předmět: |
|
| Zdroj: |
Indian journal of experimental biology. 51(11) |
| ISSN: |
0019-5189 |
| Popis: |
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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
