PRODUCTION, PURIFICATION AND CHARACTERIZATION OF LACCASE FROM WHITE ROT FUNGUS.

Abstract

:Laccases are oxidoreductases enzymes that belong to the sub class copper oxidases.
They are mainly produced by white rot fungi which are involved in lignin metabolism. The objective
of this research was the effective utilization of agro wastes for the economical production of laccase
from, fungi Pleurotusflorida. Submerged fermentation was employed for the production of enzyme
and agro wastes namely rice husk, wheat bran and sugarcane bagasse used as substrates. The activity
of enzyme was analyzed spectrophotometrically at 420nm, using ABTS (2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) as a substrate. The maximum production was obtained with sugarcane bagasse 9.9 IU/ml at day 9. The optimum temperature and pH were found to be 30˚C and 7. The best nitrogen source optimized was ammonium sulphate which enhanced the production up to
11.57 IU/ml. The large scale batch (1litre) with already optimized conditions produced 13460 IU/L of
enzyme. Purification was performed and molecular weight was determined at 90kDa through SDSPAGE. Characterization of enzyme was done in terms of pH, temperature and effect of inducers and inhibitors.