Purification and biochemical characterisation of glucoamylase from a newly isolated Aspergillus niger: relation to starch processing.

Herein, we investigate a glucoamylase from newly isolated Aspergillus niger. The enzyme was purified, using fractionation, followed by anion-exchange chromatography and then characterised. The molecular mass of the enzyme was estimated to be ∼62,000Da, using SDS-PAGE and 57151Da, based on mass spectrometry results. The pI of the protein, and optimum pH/temperature of enzyme activity were 4.4, 5 and 70°C, respectively and the kinetic parameters (Km, Vmax and kcat) were determined to be 0.33 (mgml(-1)), 0.095 (Uμg(-1)min(-1)) and 158.3 (s(-1)) for soluble starch, respectively. The glucoamylase nature of the enzyme was also confirmed using TLC and a specific substrate. Metal ions Fe(3+), Al(3+) and Hg(2+) had the highest inhibitory effect, while Ag(2)(+), Ca(2+), Zn(2+), Mg(2+) and Cd(2+) and EDTA showed no significant effect on the enzyme activity. In addition, thermal stability of the enzyme increased in the presence of starch and calcium ion. Based on the results, the purified glucoamylase appeared to be a newly isolated enzyme.