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Strain fermentation and norvancomycin yield detection

XL Xingxing Li
CZ Cong Zhang
YZ Ying Zhao
XL Xuan Lei
ZJ Zhibo Jiang
XZ Xuexia Zhang
ZZ Zhihui Zheng
SS Shuyi Si
LW Lifei Wang
BH Bin Hong
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Amycolatopsis orientalis CPCC 200066 and its derivatives were cultured in 100 ml Bennet liquid medium at 28 ℃ for 2 days, and then 10% seed culture was transferred into 100 ml fresh Bennet for continuous fermentation at 28 ℃ for 4 days. Each fermented supernatant was collected by centrifugation at 12,000 rpm for 10 min, and then filtered with microporous membrane of 0.22 μm and analyzed by HPLC–MS (Agilent 1290-Agilent 1956 single quadrupole MS coupled system). The HPLC conditions were as follows: Agilent Eclipase plus C18 column (250 mm × 4.6 mm, 5 μm), mobile phase Solvent A was 100% MeOH, Solvent B was water with 0.1% formic acid. The HPLC program included column elution with a linear gradient of 5 to 30% solvent A over 30 min at 25 ℃. The flow rate was set at 0.8 ml/min. MS spectra data were collected in the positive-ion mode in which a mass range of m/z 150 to 2,000 covered. The NVCM peak was extracted ion chromatogram (EIC) of m/z 717.9 [M + 2H]2+.

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