Bacteroides fragilis is a sphingolipid-producing bacterium. It has been hypothesized that sphingolipids allow Bacteroides fragilis to live successfully in the human gut1. The enzyme which determines the rate determining step of the sphingolipid pathway is known as serine palmitoyl transferase (SPT). Purification of BfSPT protein utilized a genetically engineered version of the BfSPT protein which includes a coded histidine tag located on the c-terminus of the protein1. After Ni-resin purification, size exclusion chromatography with a GE S200 column was utilized for further purification. The purified version of the protein had Kd =0.9998 � 0.1267mM using an UV-Vis spectrometer measuring absorption at 420nm. In DNTB assays where L-serine and palmitoyl-CoA were used as substrates, BfSPT displayed kcat = 0.167 s-1 .

<i>Bacteroides fragilis</i> serine palmitoyl transferase purification and kinetic study

247th National Meeting (2014)

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Bacteroides fragilis serine palmitoyl transferase purification and kinetic study