Journal of Pure and Applied MicrobiologyVol. 8 No. Special Edition Nov. 2014

Production and Partial Purification of Laccase from Pseudomonas aeruginosa ADN04

T. Arunkumar1*, D. Alex Anand2 and G. Narendrakumar3

1Department of Bioinformatics, Sathyabama University, Chennai - 600119, India. 2Department of Biomedical Engineering, Sathyabama University, Chennai - 600119, India. 3Department of Biotechnology, Sathyabama University, Chennai - 600119, India.

Received on 06 September 2014 and accepted on 13 October 2014

 

ABSTRACT

The efficient extracellular production of laccase in the liquid culture medium of Pseudomonas aeruginosa ADN04 and the enzyme extraction, characterization and purification were studied. Purified laccase from the culture filtrate of the bacterium has been attained maximum activity after employing ammonium sulphate precipitation, Gel filtration chromatography (Sephadex) and Affinity chromatography DEAE-Sephadex. The molecular mass of the laccase was determined by SDS-PAGE that showed a relative molecular weight of 43 kDa. The enzymatic stability characteristics at different pH and temperature of the purified laccase have been determined. 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) was used as the substrate and have been found to be Vmax is 12.06 and Km is 56.9at 37°C respectively.

Keywords : Laccase, Pseudomonas aeruginosa ADN04, Optimization, Purification