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High-yield production of maltooligosaccharide-forming α-amylase in Bacillus subtilis
Xiangyi Li1, Jianghua Yang1, Yonglu Li1
1State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Microorganism and Enzyme Research Center of Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning, Guangxi 530004, China.
Abstract:
In order to enhance the production of maltooligosaccharide-forming α-amylase (BcMFAse) in B. subtilis, we investigated the effects of the glycerol-inducible promoter PGlpD, fermentation medium optimization, homologous promoter evaluation and dual translation initiation sites (TISs) construction at 5 ´ -untranslated region (5 ´-UTR) on BcMFAse expression at the shake-flask level. The optimal induced condition was determined as inducing at 8 h with 1% glycerol. The optimized fermentation medium consisted of 2% tryptone (Tryp), 0.6% yeast extract (YE), 0.6% casein hydrolysate (CH) and 1% NaCl, yielding a BcMFAse activity of 1549.9 U/mL with promoter PGlpD. Substitution with the homologous promoter PGlpDL, resulting in an activity of 2364.6 U/mL. Construction of dual TISs under the control of PGlpDL further increased BcMFAse activity to 3137.5 U/mL. This study provides an efficient strategy for high-yield production of BcMFAse in B. subtilis.
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