Aqueous-organic two-phase systems enhance activity of undecaprenyl pyrophosphate synthase
Authors
Lidsky, Israel
ORCID
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Other Contributors
Dordick, Jonathan S.
Bystroff, Christopher, 1960-
Koffas, Mattheos A. G.
Maxwell, Patrick H.
Bystroff, Christopher, 1960-
Koffas, Mattheos A. G.
Maxwell, Patrick H.
Issue Date
2015-08
Keywords
Biology
Degree
PhD
Terms of Use
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
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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