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

Thumbnail Image
Authors
Lidsky, Israel
Issue Date
2015-08
Type
Electronic thesis
Thesis
Language
ENG
Keywords
Biology
Research Projects
Organizational Units
Journal Issue
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
Full Citation
Publisher
Rensselaer Polytechnic Institute, Troy, NY
Terms of Use
Journal
Volume
Issue
PubMed ID
DOI
ISSN
EISSN