CLONING, EXPRESSION IN E.coli AND THE ENZYMATIC PROPERTIES OF LACCASE FROM Hypsizygus ulmarius
G. Ravikumar, M. Kalaiselvi, D. Gomathi, K. Devaki and C. Uma*
ABSTRACT
In the present study a laccase from Hypsizygus ulmarius was cloned
and expressed in E.coli (BL 21). The laccase gene (1.6 Kb) which was
isolated from the mushroom Hypsizygus ulmarius was successfully
cloned to pET-29a (+) and expressed in E coli. The sequencing results
confirmed the expression of the gene with 98% homology to that of the
original gene as retrieved from NCBI. The recombinant laccase was
produced from the transformed colonies and it was purified upto 23
fold with 51% recovery. The optimum pH and temperature for the
purified laccase was found to be 6.0 and 40oC respectively. The
molecular weight of the enzyme was found to be 63kDa. The enzyme
activity was enhanced by the metal ions Mn2+and Cu2+ whereas it was
reduced by Fe2+, Na2+ and Co2+. These results suggest that the recombinant laccase purified
from the mushroom Hypsizygus ulmarius can be used for various biotechnological
applications.
Keywords: Recombinant laccase, Hypsizygus ulmarius, E.coli.
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