Amino acid analysis protocol

The cells were harvested at 36 h of the fermentation and prepared according to the methods of previous reports for amino acid analyses [15 (link), 41 (link)]. The levels of SAM, Met, Ser and Asp were measured by a Shimadzu LC10A HPLC system (Shimadzu, Kyoto, Japan) equipped with a Megres C18 column (5 μm, 4.6 mm × 250 mm) (Hanbon Sci. & Tech., China). Peak area analysis was performed based on the standard calibration curves of SAM, Met, Ser and Asp (Sangon, Shanghai, China) according to the methods of previous reports [15 (link), 44 (link), 45 ]. For the measurement of Cys, the precolumn derivatization high-performance liquid chromatographic method was performed according to previous reports [46 (link)].

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Chen H., Chai X., Wang Y., Liu J., Zhou G., Wei P., Song Y, & Ma L. (2022). The multiple effects of REG1 deletion and SNF1 overexpression improved the production of S-adenosyl-l-methionine in Saccharomyces cerevisiae. Microbial Cell Factories, 21, 174.

Publication 2022

LC10A HPLC system

Amino acid Cells Fermentation High performance liquid chromatographic

Corresponding Organization : China Pharmaceutical University

Other organizations : Taizhou University

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Variable analysis

independent variables

Time of fermentation (36 h)

dependent variables

Levels of SAM (S-Adenosylmethionine)
Levels of Met (Methionine)
Levels of Ser (Serine)
Levels of Asp (Aspartic acid)
Levels of Cys (Cysteine)

control variables

Harvesting method of cells
HPLC system (Shimadzu LC10A)
Chromatographic column (Megres C18, 5 μm, 4.6 mm × 250 mm)
Standard calibration curves for SAM, Met, Ser, and Asp
Precolumn derivatization method for Cys measurement

positive controls

Standard calibration curves of SAM, Met, Ser, and Asp

negative controls

Not explicitly mentioned

Annotations

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