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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.04 [2005] >
IJBT Vol.04(1) [January 2005] >
| Title: | Purification and characterization of catechol 1,2-dioxygenase of Pseudomonas fluorescens for degradation of 4-chlorobenzoic acid |
| Authors: | Saxena, Preti
Thakur, Indu Shekhar |
| Keywords: | catechol-1,2-dioxygenase
4-chlorobenzoic acid
chlorocatechol
DEAE-cellulose
gel filtration
SDS-polyacrylamide gel electrophoresis |
| Issue Date: | Jan-2005 |
| Publisher: | CSIR |
| IPC Code: | Int. Cl.7 A 01 N 29/00, 63/00; C 07 C 63/10 |
| Abstract: | The degradation of 4-chlorobenzoic acid (4-CBA) by Pseudomonas fluorescens IST8 was determined. The degradation of 4-CBA proceeded through an oxidative route to yield ortho ring cleavage enzyme, catechol 1, 2-dioxygenase. The cell free extract fractionated by DEAE-cellulose ion-exchange and gel filtration chromatography showed two different fractions of catechol 1,2-dioxygenase with an expected molecular weight of 62 and 48 kDa, respectively. The catechol 1, 2-dioxygenase in the fractions I and II was purified to about 22.3 and 36.5 fold by using purification steps. The pH and temperature optima for enzyme activity were 6.5 and 25°C, respectively. The purified protein on SDS-polyacrylamide gel electrophoresis showed molecular weights of 28 and 24 kDa, indicating dimeric nature of the enzyme. |
| Page(s): | 134-138 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.04(1) [January 2005]
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