Structural Characterization of NulO Pathway

For structural characterization of NulO pathway intermediates, filtered enzymatic reactions (Amicon Ultra-4 centrifugal filter unit with 3000 kDa molecular weight cut-off) or material purified according to previously described methods was exchanged into 100% D₂O (Gulati et al. 2015 (link)). Structural analysis was performed using either a Varian Inova 500 MHz (¹H) spectrometer with a Varian Z-gradient 3-mm probe or a Varian 600 MHz (¹H) spectrometer with a Varian 5 mm Z-gradient probe. All spectra were referenced to an internal acetone standard (δ_H 2.225 ppm and δ_C 31.07 ppm).

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Friedrich V., Janesch B., Windwarder M., Maresch D., Braun M.L., Megson Z.A., Vinogradov E., Goneau M.F., Sharma A., Altmann F., Messner P., Schoenhofen I.C, & Schäffer C. (2017). Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications. Glycobiology, 27(4), 342-357.

Publication 2017

Inova 500 MHz (1H) spectrometer Z-gradient 3-mm probe 600 MHz (1H) spectrometer 5 mm Z-gradient probe

Acetone Enzymatic

Corresponding Organization :

Other organizations : BOKU University, National Research Council Canada, University at Buffalo, State University of New York

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Variable analysis

independent variables

Enzymatic reactions filtered using Amicon Ultra-4 centrifugal filter unit with 3000 kDa molecular weight cut-off
Material purified according to previously described methods

dependent variables

Structural characterization of NulO pathway intermediates

control variables

Exchanging samples into 100% D2O
Using a Varian Inova 500 MHz (1H) spectrometer with a Varian Z-gradient 3-mm probe or a Varian 600 MHz (1H) spectrometer with a Varian 5 mm Z-gradient probe
Referencing all spectra to an internal acetone standard (δH 2.225 ppm and δC 31.07 ppm)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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