文献类型：期刊文献

英文题名：Production of α-cyclodextrin glycosyltransferase in bacillus megaterium ms941 by systematic codon usage optimization

作者：Zhou, Jingwen[1,2]; Liu, Hua[1,2]; Du, Guocheng[1]; Li, Jianghua[1]; Chen, Jian[1]

机构：[1] School of Biotechnology, Jiangnan University, Ministry of Education, 1800 Lihu Road, Wuxi, Jiangsu 214122, China; [2] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China

年份：2012

卷号：60

期号：41

起止页码：10285

外文期刊名：Journal of Agricultural and Food Chemistry

收录：EI(收录号：20124315603132)

语种：英文

外文关键词：Bacteriology - Cyclodextrins - Physiology - Traffic congestion - Escherichia coli

摘要：α-Cyclodextrin glycosyltransferase is a key enzyme in the cyclodextrin industry. The Gram-positive bacterium Bacillus megaterium was chosen for production of recombinant α-CGTase for safety concerns. Successful production of heterologous α-CGTase was achieved by adapting the original α-cgt gene to the codon usage of B. megaterium by systematic codon optimization. This balanced the tRNA pool and reduced ribosomal traffic jams. Protein expression and secretion was ensured by using the strong inducible promoter Pxyl and the signal peptide SPLipA. The impact of culture medium composition and induction strategies on α-CGTase production was systematically analyzed. Production and secretion at 32 °C for 24 h using modified culture medium was optimal for α-CGTase yield. Batch- and simple fed-batch fermentation was applied to achieve a high yield of 48.9 U·mL-1, which was the highest activity reported for a Bacillus species, making this production system a reasonable alternative to Escherichia coli. ? 2012 American Chemical Society.