Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from <it>Bacillus alcalophilus </it>in <it>Bacillus subtilis</it>

• Main Authors: Li Jianghua, Liu Long, Yang Haiquan, Du Guocheng, Chen Jian
• Format: Article
• Language: English
• Published: BMC 2011-10-01
• Series: Microbial Cell Factories
• Online Access: http://www.microbialcellfactories.com/content/10/1/77

<p>Abstract</p> <p>Background</p> <p>Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase.</p> <p>Results</p> <p>The alkaline α-amylase gene from <it>Bacillus alcalophilus </it>JN21 (CCTCC NO. M 2011229) was cloned and expressed in <it>Bacillus subtilis </it>strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the <it>K</it>_mand <it>V</it>_maxof alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min), respectively. The effects of medium compositions (starch, peptone, and soybean meal) and temperature on the recombinant production of alkaline α-amylase in <it>B. subtilis </it>were investigated. Under the optimal conditions (starch concentration 0.6% (w/v), peptone concentration 1.45% (w/v), soybean meal concentration 1.3% (w/v), and temperature 37°C), the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from <it>B. alcalophilus </it>JN21.</p> <p>Conclusions</p> <p>This is the first report concerning the heterologous expression of alkaline α-amylase in <it>B. subtilis</it>, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant <it>B. subtilis</it>.</p>