Tannase production by Bacillus licheniformis KBR6: Optimization of submerged culture conditions by Taguchi DOE methodology

Tannase production by Bacillus licheniformis KBR6 under submerged fermentation was optimized following Taguchi orthogonal array (OA) design of experiment (DOE). An OA layout of L18 (21 × 35) was constructed with six most influensive factors on tannase biosynthesis like, carbon source (tannic acid), phosphate source (KH2PO4), nitrogen source (NH4Cl), metal ion (Mg2+), incubation temperature and initial medium pH at three levels for the proposed experimental design. Tannase yield obtained from the 18 batches fermentation with the selected levels of each factors were processed with Qualitek-4 software at bigger is better as quality character and obtained a specific combination of factors with a predicted tannase production of 0.362 U/ml. The optimal combinations of factors (tannic acid, 1.0 g%; KH2PO4, 0.45 g%; NH4Cl, 0.35 g%; MgSO4, 0.05 g%) obtained from the proposed DOE methodology was further validated by fermentation experiment and the obtained result revealed an enhanced tannase yield of 2.18-fold (from 0.163 U/ml to 0.356 U/ml) from its unoptimized condition. Taguchi approach of DOE resulted in evaluating the main and interaction effects of the factors individually and in combination.