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Vol. 10 No. 2.0 (2021)
Scholar : National School of Leadership

Research Articles

Screening of mushrooms from exotic locales for laccase production: Studies on the enzyme from Amanita sp. on azo dye degradation.

Published 2021-12-31

  • ANANTHU A
    • ,
  • RAKHI RAJ R.S
    • ,
  • RENJU KRISHAN R.V

ANANTHU A
MAR IVANIOS COLLEGE

RAKHI RAJ R.S
MAR IVANIOS COLLEGE

RENJU KRISHAN R.V
MAR IVANIOS COLLEGE

Abstract

The present study focused on screening of various mushrooms collected from exotic locales for laccase production using SSF and SmF. The molecular identification of the potent strain was done by 18s rRNA amplification using ITS primers, ITS1(Forward) and ITS 4 (Reverse). The sequence showed 99% similarity with Amanita sp. with a query coverage of 98%. This strain was selected for further studies including partial purification, laccase immobilization and dye degradation studies.  Partial purification through ammonium sulphate precipitation showed maximum activity in 60% ammonium sulphate fraction. The extent of purification was analyzed by doing SDS-PAGE. Activity staining of non reducing SDS gel showed the presence of heterodimer having molecular weight of ~60kDa(Band 1) and 40 kDa (Band 2). Enzyme characterization of partially purified laccase was performed. Optimum temperature of laccase enzyme was found to be 50°C and optimum pH was found to be 4. The partially purified enzyme preparation from SSF was used for immobilization studies. The enzyme activity of laccase immobilized calcium alginate beads was found to be 0.8 U/10 mg. The ability of the immobilized laccase in dye degradation was analysed using 5mM methyl orange solution with 3g (~250U) of immobilized laccase. The result observed was good. An ~ 50% dye degradation was observed even in the first six hours and more than 80% of dye got degraded in 24 hours of enzyme treatment.

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