Aspergillus niger was the potent β-glucanase producer, as observed from the screening profile. Based on morphological and molecular approaches (18S-28S rRNA sequence, flanking the ITS and 5.8S rRNA regions), the isolate was identified as Aspergillus niger with accession number JQ1516491. Under submerged fermentation conditions, the maximum enzyme yield (127.3 U/mg) by A. niger was obtained using 2% CMC, 0.3% yeast extract, 0.1% KH2PO 4, 0.05% MgSO4.7H2O, 0.05% KCl. Among the tested ten natural agricultural byproducts, peanut shell cake was the potent substrate for induction of β-glucanase (178 U/mg) by A. niger under solid state fermentations (SSF). Under SSF conditions, the enzyme yield was increased by about 1.4 folds by supplementation of 0.1% CMC and 0.1% yeast extract to the same salt solution. The enzyme was purified and characterized from the solid cultures of A. niger using peanut as substrate, by salting out, gel-filtration and ion exchange chromatography. Following the chromatographic step, the enzyme activity was increased by 5.4 fold with 42.6% yield. Using SDS-PAGE, the enzyme has molecular weight 55 and 35 kDa. The maximum enzyme activity was measured using 1% β-glucan in potassium phosphate buffer of pH 6.5-7.0, with relative pH stability at 5.4 to 6.0. Also, the highest enzyme activity was detected by incubation of the reaction mixture at 50°C, with plausible thermal stability at this degree. The enzyme thermal denaturation rate was increased subsequently with the heating temperature, as revealed from the T 1/2 values that were 10, 4.7 and 2.7 hr at 50, 60 and 70°C, respectively. The enzyme has iso-electric focusing (pI) at pH 6.6-7.4. From the absorption spectra, the enzyme has a distinct peak at 230 nm for the apo- enzyme and other at 300 nm.