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thereafter, 50 ml of precultures were induced with 450 ml of expression medium (10 g l 1 yeast extract (Gibco, 212750), 20 g l 1 bacto peptone (Gibco, 211677), 20 g l 1 d-galactose (Panreac, A1131), 67 mM potassium phosphate monobasic buffer pH 6.0 (Thermo Scientific, 424200250), 25 g l 1 ethanol (Scharlau, ET00062500), 2 mM copper sulfate (Merck, C7631) and 25 g ml 1 chloramphenicol (Merck, C0378)) in a 2.5-l baffled flask (Quimigen, 931136-B) (OD 600 = 0.1)