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Benzoate

Manufactured by Merck Group
Sourced in Sao Tome and Principe

Benzoate is a chemical compound used in various laboratory applications. It is a salt or ester of benzoic acid, which is a naturally occurring organic compound. Benzoate serves as a preservative, pH regulator, and intermediate in organic synthesis. The specific details and intended use of this product may vary, and we recommend consulting with our technical specialists for more information.

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4 protocols using benzoate

1

Characterization of Bacterial Gene Circuits

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For each circuit separate colonies of E. coli TOP10 (strain K-12, genotype: FmcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG) strains harboring the circuit plasmids were cultured overnight at 37 °C in LB with appropriate antibiotic. The next day each culture was diluted 100× in LB with antibiotics. 95 µL of fresh cultures were distributed in 96-well plate (Corning 3603) and the plate was incubated to reach the OD600 ~ 0.1 in a plate reader (Biotek Synergy HTX). Then 5 µL of the input metabolites (100× ethanol solutions 5x diluted in LB) were added and the plate was incubated for 18 h at 37 °C. During the incubation, the OD600 and GFP fluorescence (gain: 35, ex: 458 nm, em: 528 nm) were measured. Benzoate, hippurate, cocaine hydrochloride, benzaldehyde, benzamide and biphenyl-2,3-diol (2,3-dihydroxy-biphenyl) were purchased from Sigma-Aldrich. Permission to purchase cocaine hydrochloride was given by the French drug regulatory agency (Agence Nationale de Sécurité du Médicament et des Produits de Santé). For all chemicals, serial dilutions of 100× concentrations were prepared in ethanol. The formula presenting the results of the circuits’ characterization is shown in data normalization section. The mean and standard deviation of all normalized data are provided in Supplementary Table 7.
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2

Comprehensive Metabolomics Sample Preparation

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Methanol, acetonitrile, perchloric
acid (PCA), trichloroacetic
acid (TCA), hydrochloric acid (HCl), sodium hydroxide (NaOH), (2-bromoethyl)
trimethylammonium bromide, dimethylformamide (DMF), 3-(trimethylsilyl)propionic
acid-2,2,3,3-d4 sodium salt (TSP) were
all obtained from Sigma-Aldrich (St. Louis, MO). 4-(4,6-Dimethoxy[1,3,5]triazin-2-yl)-4-methylmorpholinium
chloride (DMTMM) was obtained from Acros Organic (Pittsburgh, PA),
while 15N-phthalimide potassium and deuterium oxide were
obtained from Cambridge Isotope Laboratories (Andover, MA). All chemicals
were used without further purification. Pooled human serum sample
was obtained from Innovative Research, Inc. (Novi, MI). Deionized
(DI) water was purified using an in-house Synergy Ultrapure Water
System from Millipore (Billerica, MA). Centrifugal filters (3-kDa
cutoff; Amicon Microcon, YM-3) were purchased from Sigma-Aldrich.
Standard compounds used for spiking and confirming the peak assignments
were 1-methylhistidine, 2-hydroxybutyrate, 2-hydroxyisocaproate, 2-hydroxyisovalerate,
2-oxocaproate, 3-methylbutyrate, 3-methylhistidine, 3-methyl-2-oxobutanoate,
arginine, benzoate, betaine, carnitine, citrulline, dimethylglycine,
pyridoxine, pyroglutamate, ornithine, sarcosine, serine and succinate
(all from Sigma-Aldrich).
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3

Analytical Protocol for Organic Compounds

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Benzoate, 3-hydroxyBenzoate, 4-hydroxyBenzoate, cinnamate, 3-hydroxycinnamate, salicylate, nicotinate, anthranilate, 4-aminoBenzoate, 2-aminophenol, benzamide, simazine, tryptophan, and methyl viologen dichloride hydrate (paraquat, PQ) were obtained from Sigma-Aldrich (St. Louis, MO, United States). Benzene (>99.7% purity), toluene, biphenyl, naphthalene, phenanthrene, anthracene, fluorene, and 30% (w v-1) hydrogen peroxide (H2O2) were purchased from Merck (Darmstadt, Germany).
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4

Vibrational Spectroscopy of Sodium Carboxylates

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We used sodium carboxylates as received:
formate (≥99.0%, Fluka Analytical), acetate (anhydrous, for
molecular biology ≥99%, Sigma-Aldrich), propionate (minimum
99%, Sigma), hexanoate (99–100%, Sigma), octanoate (≥99%,
Sigma), benzoate (>99%, Sigma-Aldrich), and 2-naphthoate (>98%,
TCI).
To prepare the samples, we dissolved appropriate amounts of the salts
in D2O (99.9% D atom, Aldrich). For each carboxylate, we
chose the concentration to obtain a signal-to-noise ratio that is
more than sufficient to reliably determine the amplitudes of the VSFG
responses of the νs and νas vibrations
of the carboxylate groups in different VSFG polarization combinations.
The concentrations of the solutes are reported in the units of molality
(mol/kg solvent (m)).
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