Quantitative Amino Acid Analysis in Bacterial Cells
Corresponding Organization :
Other organizations : Kobe University, Nara Institute of Science and Technology, Osaka University, Kao Corporation (Japan), Tokyo Institute of Technology, Toyama Prefectural University
Variable analysis
- Amount of culture collected (OD600 × culture volume (ml) = 8)
- Mass isotopomer distributions (MIDs) of proteinogenic amino acids
- Culture collected during the exponential phase (7.5–8 h)
- Washing with 0.2 mol l−1 HCl to remove CaCO3
- Washing with saline
- Hydrolysis with 6 mol l−1 HCl at 105°C for 18 h
- Filtration through a Cosmonice filter W (0.45 µm)
- Addition of 10 µl of 600 µM cycloleucine
- Drying and dissolution in 50 µl acetonitrile
- Derivatization with N-(tert-tert-butyldimethylsilyl)-N-methyl-trifluoroacetamide containing 1% tert-butyldimethylchlorosilane at 105°C for 1 h
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