Products/Services Used | Details | Operation |
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DNA Sequencing> | … Co., Ltd., Nanjing, China). The recombinant plasmid was transformed into E. coli BL21 (DE3) and designated as E. coli BL21 (DE3-pET-opt-cgt). Finally, the insertion of the genes was confirmed by DNA sequencing (GenScript Biotech. Co, Ltd., Nanjing, China) … | Get A Quote |
Background Cyclodextrin glucanotransferase (CGTase) can transform L-ascorbic acid (L-AA, vitamin C) to 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), which shows diverse applications in food, cosmetic and pharmaceutical industries. Results In this study, the cgt gene encoding α-CGTase from Paenibacillus macerans was codon-optimized (opt-cgt) and cloned into pET-28a (+) for intracellular expression in E. coli BL21 (DE3). The Opt-CGT was purified by Ni2+-NTA resin with a 55% recovery, and specific activity was increased significantly from 1.17 to 190.75 U·mg− 1. In addition, the enzyme was adopted to transform L-AA into 9.1 g/L of AA-2G. Finally, more economic substrates, including β-cyclodextrin, sol... More