Vivaspin 20 spin filters

Lysogeny broth (LB) medium, isopropyl β-d-thiogalactopyranoside (IPTG), and NADPH were purchased from
Research Products International. The protease inhibitor cocktail,
lysozyme, DNase I, acetaldehyde dehydrogenase, pyrophosphatase, d-fructose-6-P, d-fructose-1-P, d-glucose-6-P, d-glucose-1-P, d-glucitol-6-P, CTP, UTP, ATP, GTP,
NADPH, NADH, acetaldehyde, kanamycin, imidazole, and HEPES were obtained
from Sigma-Aldrich. Vivaspin 20 spin filters and HisTrap and HiTrap
Q columns were obtained from Cytiva. The 10 kDa Nanosep spin filters
were purchased from Pall Corp. (Port Washington, NY). Deuterium oxide
was acquired from Cambridge Isotope Laboratories Inc.

All
materials used in this study were obtained
from Sigma-Aldrich, Carbosynth, or GE Healthcare Biosciences, unless
otherwise stated. Lysogeny broth (LB) and isopropyl-β-d-thiogalactopyranoside (IPTG) were purchased from Research Products
International. HisTrap columns and Vivaspin 20 spin filters were obtained
from Cytiva. The 3 and 10 kDa Nanosep spin filters were purchased
from Pall Corporation (Port Washington, NY). Deuterium oxide was purchased
from Cambridge Isotope Laboratories, Inc. Escherichia coli strain BL21-Gold (DE3) was obtained from New England Biolabs. α-Ketoglutarate
was purchased from AK Scientific (Union City, CA). Ultraviolet spectra
were recorded on a SpectraMax340 (Molecular Devices) ultraviolet–visible
plate reader using 96-well Greiner plates. ¹H NMR and ¹H–¹H COSY spectra were recorded on a Bruker
Avance III 400 MHz system equipped with a broadband probe and sample
changer. Mass spectrometry data were collected on a Thermo Scientific
Q Exactive Focus system run in the negative-ion mode. GDP-d-glycero-α-d-manno-heptose (5) was synthesized and purified as described
previously.^{20 (link)}