4 aminobenzoic acid

Reagents and solvents were obtained from Sigma-Aldrich (glutaric
acid, isophthalic acid, trimesic acid, hydroquinone, benzoquinone,
4-aminobenzoic acid, oxalic acid, 1-naphthol, terephthaldehyde, saccharin,
nicotinamide, resorcinol, and 2,6-pyridinecarboxylic acid) and TCI
(benzoic acid, trans-1,2-bis-(4-pyridyl)ethylene, 4,4′-bipyridine,
1,2-bis-(4-pyridyl)ethane, 4,4′-biphenol, tetramethylpyrazine,
naproxen, carbamazepine, aspirin, and 2,7-dihydroxynaphthalene) and
used as received. X-ray powder diffraction (PXRD) was used for phase
identification of coformers and cocrystals. We note that peak positions
in experimental PXRD patterns can exhibit slight shifting compared
to calculated PXRD patterns obtained from SCXRD data because of thermal
expansion if data were collected at different temperatures.

Titanium(IV) isobutoxide (Aldrich, St. Louis, MO, USA), 4-aminobenzoic acid (Aldrich, St. Louis, MO, USA), 9-fluorenecarboxylic acid (Organic Acros, Geel, Belgium) were purchased commercially and were used without further purification. All solvents used in synthesis, i.e., acetone and toluene were distilled before their use and stored in an argon atmosphere. The processes of Ti(IV) oxo-complexes synthesis were carried out using the standard Schlenk technique in the inert gas atmosphere (Ar) and at room temperature (RT).

Titanium(IV) isopropoxide (Aldrich, St. Louis, MO, USA), titanium(IV) isobutoxide (Aldrich, St. Louis, MO, USA), 4-aminobenzoic acid (Aldrich, St. Louis, MO, USA), and 4-hydroxybenzoic acid (Aldrich, St. Louis, MO, USA) were purchased commercially and were used without further purification. All solvents used in the synthesis, i.e., tetrahydrofuran (THF), isobutanol (HOⁱBu), and isopropanol (HOⁱPr) were distilled before their use and stored in argon atmosphere. The processes of Ti(IV) oxo-complexes synthesis were carried out using the standard Schlenk technique in the inert gas atmosphere (Ar) and at room temperature (RT).

The redox state of roGFP2 in PLB-985 was measured in a 96-well format as described by Degrossoli et al. with minor modifications [37 ]. In short, 50 μl of roGFP2 expressing PLB-985 cells at a concentration of 10⁷ cells/ml were incubated with respective inhibitors as described (100 nM Wortmannin; 10 μM Diphenyleneiodonium chloride (DPI); 500 μM 4-aminobenzoic acid hydrazide (ABAH); 1 μM Gö 6983 (Gö)) (Sigma, Darmstadt, DE) or in case of control, with PBS for 1 h at 37 °C in a 96-well plate (Nunc black, clear-bottom, Rochester, NY). Afterwards, 50 μl of E. coli at an OD₆₀₀ of 1.0 as well as the respective stimulants were added. The fluorescence intensity was recorded every minute for 2 h at the excitation wavelength 405 nm and 488 nm, unless described otherwise. The emission wavelength was set to 510 nm. The Calculation of the 405/488 nm ratio was done using Microsoft Excel 2016 (Microsoft, USA). Visualization of respective graphs were done using GraphPad Prism (version 5.00, USA). All plate reader assays were performed in at least three independent experiments. The end point ratio, as depicted in the bar graphs, was calculated based on the final point of a linear regression over the last 10 min of the measurement.

L-glutathione reduced (GSH), 4-aminobenzoic acid (ABA), N-hydroxysulfosuccinimide (NHS), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), 2-(N-morpholino)-ethanesulfonic acid (MES), sodium nitrite, hydrochloric acid (HCl), potassium ferrocyanide K₄[Fe(CN)₆]·3H₂O, potassium ferricyanide K₃[Fe(CN)₆], iron(II) sulfate heptahydrate (99%), potassium chloride (99%), 30 wt% hydrogen peroxide solution, glutathione reductase from baker’s yeast (S. cerevisiae), and nicotinamide adenine dinucleotide phosphate (NADPH) were purchased from Sigma–Aldrich (USA). A screen-printed carbon electrode serving as a transducer for the sensor base was obtained from Pine Instruments, USA. Each electrode consists of a carbon-working electrode with a diameter of 2 mm, a silver/silver chloride reference electrode, and a carbon auxiliary electrode. A Gamry Reference 600 potentiostat (Gamry Instruments, USA) was used to conduct electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) measurements.

Neisseria gonorrhoeae was grown overnight at 37°C and 5% CO₂ on agar plates containing gonococcal base (GC) agar [10 g/l Bacto agar (BD Biosciences, Bedford, MA, United States), 5 g/l NaCl (Roth, Darmstadt, Germany), 4 g/l K₂HPO₄ (Roth), 1 g/l KH₂PO₄ (Roth), 15 g/l Proteose Peptone No. 3 (BD), 0.5 g/l soluble starch (Sigma-Aldrich, St. Louis, MO, United States)] supplemented with IsoVitaleX (IVX): 1 g/l D-Glucose (Roth), 0.1 g/l L-glutamine (Roth), 0.289 g/l L-cysteine-HCL × H₂0 (Roth), 1 mg/l thiamine pyrophosphate (Sigma-Aldrich), 0.2 mg/l Fe(NO₃)₃ (Sigma-Aldrich), 0.03 mg/l thiamine HCl (Roth), 0.13 mg/l 4-aminobenzoic acid (Sigma-Aldrich), 2.5 mg/l β-nicotinamide adenine dinucleotide (Roth) and 0.1 mg/l vitamin B₁₂ (Sigma-Aldrich). GC medium is identical to the base agar composition but lacks agar and starch.
E. coli was grown in LB (Lysogeny Broth, Roth) medium or on LB agar plates (15 g/l Bacto agar (BD Biosciences, Bedford, MA, United States) at 37°C.
For N. gonorrhoeae antibiotics were used at the following concentrations: 2.5–5 μg/ml erythromycin (Thermo-Fisher), 100 μg/ml streptomycin (Sigma-Aldrich), 10 μg/ml chloramphenicol (Sigma-Aldrich). For E. coli antibiotics were used at the following concentrations: 50 μg/mL kanamycin (Roth).