Aqueous-organic two-phase systems enhance activity of undecaprenyl pyrophosphate synthase

Authors
Lidsky, Israel
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Other Contributors
Dordick, Jonathan S.
Bystroff, Christopher, 1960-
Koffas, Mattheos A. G.
Maxwell, Patrick H.
Issue Date
2015-08
Keywords
Biology
Degree
PhD
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This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.
Full Citation
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
Department
Dept. of Biological Sciences
Publisher
Rensselaer Polytechnic Institute, Troy, NY
Relationships
Rensselaer Theses and Dissertations Online Collection
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