Lipoteichoic acid is an important microbe-associated molecular pattern of Lactobacillus rhamnosus GG

Background

Receptors with a single transmembrane (TM) domain are essential for the signal transduction across the cell membrane. NMR spectroscopy is a powerful tool to study structure of the single TM domain. The expression and purification of a TM domain in Escherichia coli (E.coli) is challenging due to its small molecular weight. Although ketosteroid isomerase (KSI) is a commonly used affinity tag for expression and purification of short peptides, KSI tag needs to be removed with the toxic reagent cyanogen bromide (CNBr).

Result

The purification of the TM domain of p75 neurotrophin receptor using a KSI tag with the introduction of a thrombin cleavage site is described herein. The recombinant fusion protein was refolded into micelles and was cleaved with thrombin. Studies showed that purified protein could be used for structural study using NMR spectroscopy.

Conclusions

These results provide another strategy for obtaining a single TM domain for structural studies without using toxic chemical digestion or acid to remove the fusion tag. The purified TM domain of p75 neurotrophin receptor will be useful for structural studies.     

Enhanced system kLa and permeate flux with a ceramic membrane bioreactor

Summary A cross-flow ceramic membrane was coupled to a bioreactor to fulfil the alternate functions of process stream clarifier and primary aerator. At the same air supply rate (delivered as a back-flush), ceramic membranes provided up to 72% greater aeration than a ring-sparger located in the bioreactor. In Biol.ogical treatment of dairy food process waste, the air backflush had the additional benefit of inhibiting membrane fouling, thereby maintaining higher (by about 100%) permeate fluxes.