Team Nanjing_NFLS 2020 use this part to increase the rhamnose synthesis and upregulate PYO level in Pseudomonas aeruginosa, which contributes to improvement of our efficiency of Microbial fuel cells (MFCs) this year. We added rhlA on the plasmid pBBR1MCS-5. Restriction endonuclease digestion result (Figure 1) showed the gene is successfully bind to the plasmid.
Figure 1. Identification of pBBR1MCS-5-phzM by restriction endonuclease digestion
We then tested the efficiency of the enzyme. First, different concentrations of rhamnose standard solution was prepared, and the absorbance was determined at 620 nm at different concentrations. Then the standard curve shown as y = 0.0065 x + 0.0081 (R2=0.9993), y for the absorbance of the sample, and x for the concentration of rhamnose in the corresponding sample(mg/L). According to the standard curve, rhamnose was 44.21% higher in engineered cells than that of the control group(Figure 2).
strain
rhamnose concentration (mg/L)
PAO1
409.80
PAO1-phzM
590.96
Figure.2 Rhamnose analysis in overexpression rhlA in P. aeruginosa PAO1
