Fig. 1
Stress response induced by secretion of cutinase fused to different signal peptides. A Growth and secretion stress were measured online in a Biolector microbioreactor. B. subtilis PAL5 harboring the expression vectors pBS-4nt-SPPel-Cut11 (SPPel), pBS-4nt-SPEpr-Cut11 (SPEpr) or pBS-4nt-SPBsn-Cut11 (SPBsn) for cutinase secretion or the empty vector (EV) was grown for 24 h and mCherry fluorescence (Ex. λ = 580 nm, Em. λ = 610 nm) as well as cell density (insert: light scattering at λ = 620 nm) were determined. Expression vectors harbor the constitutive PHpaII promoter, a four nucleotide long ribosome binding site spacer between Shine-Dalgarno sequence and start codon and the GFP11 tag encoding sequence at the 3’-end of the cut gene. B Enzymatic activities in supernatants from BioLector grown cultures were determined with pNPP as substrate. Cultivation was carried out in biological triplicates and hydrolytic activity was measured in technical triplicates. Error bars indicate the standard deviations