Potato flour mediated solid-state fermentation for the enhanced production of Bacillus thuringiensis-toxin

作者：Smitha Robinson Babysarojam; Jisha Veloorvalappil Narayanan; Pradeep Selvanesan; Josh Moolakkariyil Sarath; Benjamin Sailas^*

来源：Journal of Bioscience and Bioengineering, 2013, 116(5): 595-601.

DOI：10.1016/j.jbiosc.2013.05.008

• Bacillus thuringiensis subsp kurstaki
• Solid-state fermentation
• Potato flour supplement
• Endospore
• delta-Endotoxin
• SIDS-PAGE

In this study, we explored the efficacy of raw potato flour (PF) as supplement to the conventional LB medium (LB control, designated as M1) for enhancing the concomitant production of endospores and delta-endotoxin from Bacillus thuringiensis subsp. kurstaki by solid-state fermentation (SSF). Of different concentrations and combinations of media tested, 10% (wilt) PF supplemented LB medium (M2) was found as the best source for the maximum yield of toxin. After 12 h submerged fermentation (SmF) at 37 degrees C and 125 rpm, M2 was made into a wet-solid matter for SSF by removing the supernatant (1000 xg, 10 min); the resultant pellet subsequently incubated statically (37 C) for the production of B. thuringiensis subsp. lcurstaki toxin (Btk-toxin). In comparison to Ml, yield of 3-endotoxin purified by sucrose density gradient centrifugation method from M2 was about 6-fold higher (53% recovery). This maximum yield from M2 was obtained at 48 h (as against 72 h from M1), thus the gestation period of M2 was reduced by 24 h with higher yield. In addition to the quantitative data, qualitative photomicrographs taken by image analyzer, scanning electron and fluorescent microscopes and digital camera showed physical evidences for the upper hand of SSF over conventional SmF for the enhanced production of Btk-toxin. SOS-PAGE image of the purified d-endotoxin showed three major fractions with apparent MWs 66, 45 and 30 liDa. Briefly, if low-cost agricultural products like PF is used as supplement to LB, by SSF strategy, production of Btk-toxin could be enhanced to 6-fold in short gestation time without losing its entomotoxicity efficiency.

• 出版日期2013-11