The Presence of Modifiable Residues in the Core Peptide Part of Precursor Nisin Is Not Crucial for Precursor Nisin Interactions with NisB- and NisC
Figure 2
Co-purification assay of the nisin modification enzymes NisB and NisC with different precursor nisin variants.
2A. SDS-PAGE analysis of co-purification assay of strains expressing wild type enzymes NisBTC and: Lane 1: precursor nisin NisA-H6; Lane 2: Cys-less precursor nisin (NisA-H6 C-less); lane 3: Ser/Thr-less precursor nisin (NisA-H6-TS); Lane 4: empty vector (negative control). 2B. SDS-PAGE analysis of co-purification assay of strains expressing wild type enzymes NisBTC and: Lane 1: NisA-H6 T2D P9E; lane 2: NisA-H6 T2D P9E K12D N20E; lane 3: NisA-H6 T2D P9E K12D N20E H27D K34E. 2C. Western blot of co-purification assay of strains expressing wild type enzymes NisBTC and precursor nisin variants. Co-purified NisC was detected by anti-NisC antibodies. Lane 1: Cys-less precursor nisin (NisA-H6 C-less); lane 2: Ser/Thr- less precursor nisin (NisA-H6 TS-less). The lower band represents NisC, the upper band represents NisC in a complex with NisA.