Optimization of Aspergillus niger Fermentation for the Production of Glucose Oxidase

A number of nutritional factors influencing glucose oxidase (EC 1.1.3.4) production by Aspergillus niger NCIM 545 were studied. The synthesis of glucose oxidase by A. niger was investigated in two steps using submerged fermentation at 30±2 °C and 180 rpm for 96 h. Primarily, nutritional components were selected by onefactor-at-a-time method, and the significance of each component with respect to glucose oxidase production was identified by Plackett–Burman design (seven variables including six nutritional viz. sucrose, sodium nitrate, peptone, calcium carbonate, magnesium sulfate, and potassium dihydrogen phosphate, and one dummy or unassigned variable were studied with eight experiments). In the second step, concentration of most significant factors and their interaction were studied with response surface methodology (central composite design). Each variable in the design was studied at five different levels, with all variables taken at a central coded value of zero. Considerable amount of glucose oxidase was produced from A. niger species with sucrose as the carbon source, sodium nitrate as the inorganic nitrogen source, and peptone as the organic nitrogen source. Glucose oxidase activity increased remarkably by 28.93 fold (from 0.00993 to 0.29 U ml) with CaCO3-supplemented media. The outcome of Plackett– Burman design showed CaCO3, peptone, and MgSO4 as significant parameters. Further optimization using a threefactor central composite design with 20 experiments increased yield of glucose oxidase from 0.29 to 2.05 U ml (sevenfold) with a decrease in cultivation time from 96 to 72 h.