Journal: Metabolic Engineering
Article Title: Establishing recombinant production of pediocin PA-1 in Corynebacterium glutamicum
doi: 10.1016/j.ymben.2021.09.002
Figure Lengend Snippet: (A) Growth inhibitory activity in different dilutions of fractions F8-10 of CIEX chromatography of precipitated supernatant of C. glutamicum CR099/pEKEx- pedACD Cg grow in 2xTY medium with 2% Glc in non-baffled Erlenmeyer flasks o/N. Samples were analyzed untreated or following incubation with proteinase K (+protK). All values are mean ± standard deviation of n = 3 independent experiments. The red and green lines indicate OD 600 at baseline (i.e. complete inhibition of growth) or in the absence of AMPs, respectively. The broken red line represents growth inhibition of 50%, i.e. the threshold to calculate bacteriocin units. (B) SDS-PAGE of CIEX peak fractions F9 (IEX F9) of supernatants of C. glutamicum CR099/pEKEx- pedACD Cg (CR099 pedACD Cg ) or P. acidilactici 347 ( P. aci ). Pure pediocin (PA-1) was analyzed on the same gel as control (M: molecular weight marker). (C) SIEX chromatography of pooled CIEX fractions F8–F10 from supernatants of P. acidilactici 347 ( P. aci , middle panel) or C. glutamicum CR099/pEKEx- pedACD Cg (CR099 pedACD Cg ; lower panel) or pure pediocin (PA-1) as a control. The red box indicates a signal in all samples with a size corresponding to pure pediocin. (D) Mass spectrometry of active fractions after RPC of pediocin PA-1 (upper panel) and pooled active CIEX fractions from supernatants C. glutamicum CR099/pEKEx- pedACD Cg (lower panel) before applying to RPC. The red and black boxes highlight peaks observed in both samples with almost identical mass/charge ratio (m/z) that match the calculated m/z of pediocin PA-1.
Article Snippet: For identification, the eluate from CIEX chromatography was applied to a Resource reverse-phase chromatography (RPC) column (1 ml) (GE Healthcare Biosciences) connected to an ÅKTA purifier system (Amersham Pharmacia Biotech).
Techniques: Activity Assay, Chromatography, Incubation, Standard Deviation, Inhibition, SDS Page, Molecular Weight, Marker, Mass Spectrometry
Cytiva Europe is a verified supplier 
Cytiva Europe manufactures this product 
