Production, prufication and characterization of lipase from Aspergillus niger HBF 39

Extracellular lipase produced by Aspergillus niger HBF 39 was purified 11 fold with a recovery of %35 referred to lipase activity in the crude extract using, DEAE Sepharose CL- 6B, Butyl Sepharose 4 Fast Flow, Q Sepharose Fast Flow of the purified enzyme gave a single stained band at a molecular mass of approximatly 67.5 kDa. The temperature and pH for maximum activity of the enzyme were 300C and 6.0 respectly. Km and Vmax values for pNPL of the lipase enzyme were calculated to be 49 μM and 139 U/mL, respectively. The lipase activity was stimulated by Ba2+, Li+, Na+ and K+ cations but inhibited by Fe3+, Pb2+ and Hg2+ cations. The enzyme activity was inhibited in the presence of NBS, DTT, DNTB, PMSF, CMC ve β-mercaptoethanol. These results shows that tryptophan, serine, cysteine, and carboxyl grup residues play an important role in the catalytic process. The lipase exhibited broad substrate specificity.