Aqueous-organic two-phase systems enhance activity of undecaprenyl pyrophosphate synthase
Authors
Lidsky, Israel
Issue Date
2015-08
Type
Electronic thesis
Thesis
Thesis
Language
ENG
Keywords
Biology
Alternative Title
Abstract
Typically, CPTs or their products are membrane-associated, as opposed to their hydrophilic IPP and FPP substrates. Surfactant may be interacting with UppS in a manner similar to in vivo lipid bodies, as the AOTPS has hydrophilic and hydrophobic solvents interacting at an interface. We hypothesize that an aqueous-organic two-phase system (AOTPS) with equal parts aqueous buffer and organic solvent will mimic in vivo lipid bodies and allow for CPT activity. We employed liquid-chromatography mass-spectrometry to measure substrate consumption and UPP formation from non-derivatized samples. The research presented herein shows that a UppS AOTPS reaction cannot fully replace the requirement for surfactants, but demonstrates that an AOTPS can enhance product-formation for reactions containing surfactant. We also show that different organic solvents in an AOTPS will have an effect on UppS product formation. Liquid-chromatography mass-spectrometry is used as the method for measuring substrate consumption and UPP formation from non-derivatized samples. These data suggests that UppS is a good model for studying CPT activity, which may be used as a surrogate model for studying rubber transferases.
Description
August 2015
School of Science
School of Science
Full Citation
Publisher
Rensselaer Polytechnic Institute, Troy, NY