Fig. 5

Expressional profiling of lin genes and enzyme activities of the ligase LinA and the transferase LinB. A Expressional profiling of linA and linB in L. pennispora grown in meat and PDB medium. The expression was referenced to growth in Aspergillus minimal medium and normalized against the housekeeping gene encoding the glyceraldehyde-3-phosphate dehydrogenase (gpdA). Both genes were co-expressed. B Coupled multi-enzyme assay to determine the substrate specificity of LinA. C Substrate profiling of LinA. All substrates were used at a final concentration of 1 mM. Water served as negative control. The assay was performed as described [49]. D LinB activity assay. LinB was incubated with IAA-CoA and either AA to produce 4 or HAA to produce 5. Overlaid extracted ion chromatograms (EIC) were shown for 4 (m/z 293 [M−H]⁻, in red) and 5 (m/z 309 [M−H]⁻), in blue), respectively. Assays without enzyme or without substrates were included as negative controls. Authentic standards of 4 and 5 served as positive controls