Use of surface response methodology to optimize culture conditions for iturin A production by Bacillus subtilis in solid-state fermentation

Abstract

Response surface methodology (RSM) was employed to optimize the cultivation conditions of Bacillus subtilis S3 for the enhancement of iturin A, a lipopeptide antibiotic used as biological pesticide, production in solid-state fermentation (SSF). The statistic experimental model predicted a maximum iturin production of 11.435 mg/g-wet solid material. Verification of the calculated maximum was done with experiments that were performed in the culture media representing the optimum combination found, and the iturin A production of 11.447 mg/g-wet solid material (average of three repeats) was obtained when B. subtilis S3 was cultivated at 25 °C for 5 days in solid fermentation containing high gluten flour 10 g and rice bran 50 g in addition to glucose 1.15%, KH₂PO₄ 1.27 mM, MgSO₄ 5.08 mM, peanut oil 1.01%, inoculum 19.49% and water content 44.97%. The iturin A production by B. subtilis S3 was increased significantly by 23%, from 9.26 mg/g-wet solid material to 11.447 mg/g-wet solid material when the strain was cultivated in the optimal medium developed by surface response methodology, as compared to medium conventionally developed by one-factor-at-a-time. The yield of iturin A (11.447 mg/g-wet solid material, with 45% moisture content) produced by B. subtilis S3 reported in this study is the highest reported to date for B. subtilis species in SSF. In addition, the use of rice bran as a substrate in solid-state fermentation for iturin A production by B. subtilis is unique.