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WANG Yuanxi, CHEN Junsheng, SHAO Lei, LI Ji′an, LIN Huimin, CHEN Daijie. Heterologous expression and in vitro activity of vancomycin glycosyltransferase GtfEJ. Journal of China Pharmaceutical University, 2013, 44(3): 272-276. DOI: 10.11665/j.issn.1000-5048.20130317
Citation: WANG Yuanxi, CHEN Junsheng, SHAO Lei, LI Ji′an, LIN Huimin, CHEN Daijie. Heterologous expression and in vitro activity of vancomycin glycosyltransferase GtfEJ. Journal of China Pharmaceutical University, 2013, 44(3): 272-276. DOI: 10.11665/j.issn.1000-5048.20130317

Heterologous expression and in vitro activity of vancomycin glycosyltransferase GtfE

  • Recombinant protein of vancomycin glycosyltransferase, GtfE, was obtained by gene cloning and hete-rologous expression. Gene gtfE was amplified from the genomic DNA of vancomycin producing strain and ligated into expression vector pET-37b, the recombinant plasmid GtfE/pET-37b was transformed into E. coli BL21(DE3). The product of in vitro glucosylation reaction catalyzed by purified GtfE was isolated and identified by HPLC and ESI-MS, respectively. The assay indicated that the recombinant protein GtfE had expected glucosylation activity to transfer uridine 5′-diphosphoglucose to the vancomycin aglycone. This study laid the foundation for producing glycosyl diversity of vancomycin.
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