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lactis NZ9000 harboring plasmids pNZ-USP-TAT-GFP or pNZ-USP-GFP were diluted (1:50) in fresh GM-17 medium supplemented with chloramphenicol, and grown under constant agitation at 30 C for 68 h to reach the stationary phase, corresponding to an optical density of 1.8-2 measured at a wavelength of 600 nm (OD 600 ) using a GENESYS 10UV Spectrophotometer (Thermo Scientific Inc., Waltham, MA, USA)