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Co-incubation of 4 and 11 in E. coli expressing flsQ1

CH Chunshuai Huang
CY Chunfang Yang
WZ Wenjun Zhang
LZ Liping Zhang
BD Bidhan Chandra De
YZ Yiguang Zhu
XJ Xiaodong Jiang
CF Chunyan Fang
QZ Qingbo Zhang
CY Cheng-Shan Yuan
HL Hung-wen Liu
CZ Changsheng Zhang
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The flsQ1 gene was amplified by PCR from the genomic DNA of M. rosaria SCSIO N160, and was cloned into pET28a to afford pCSG5209 (Supplementary Tables 1 and 2). Expression of flsQ1 was induced by the addition of 0.1 mM isopropyl-β-d-thiogalactopyranoside (IPTG) into E. coli BL21(DE3)/pCSG5209 growing at 16 °C to an A600 of around 0.7. After further incubation for 3 h, both compounds FST C (4) and prefluostatin (11) were added to the culture for additional incubation of 20 h. The products were then extracted with butanone and analyzed by the HPLC. E. coli BL21(DE3) harboring pET28a was treated in the same manner as a control.

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