Abstract:
NADPH dependent 1,3,6,8- tetrahydroxynaphthalene Reductase, one
of the key enzymes of 1,8-dihydroxy melanin (DHN melanin) biosynthesis
was isolated from verticillium dahliae brm-I (ATCC 44571) and purified to
near homogeneity (as shown by SDS-PAGE), using ammonium sulphate
fractionation, ion exchange chromatography on DEAE Cellulose, FPLC
Mono Q anion exchange and Superose - 12 gel filtration chromatography,
Fold purification and yield of the enzyme were found to be 1423 and 0.47%
respectively, Specific activity ofthe final enzyme fraction was 79,7 (nmole/
mg.min) Approximate molecular weight of the purified enzyme was determined
as-24 kDa by Superose-12 gel filtration chromatography,
A satisfactory HPLC based enzyme assay was also developed for thepurification of enzyme
