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glutamicum cells from pre-cultures were washed once with TN buffer pH 6.3 (10 mM Tris–HCl, 150 mM NaCl), inoculated to an optical density at 600 nm (OD 600 ) of 1 in CGXII minimal medium (Eggeling and Bott, 2005) with 40 g L –1 glucose as sole carbon source and incubated at 30°C and 120 rpm (shaking diameter 16.5 cm)