Ethylbenzene

Zeocin was purchased from Invitrogen
(USA). The Escherichia coli strain
XL2-Blue competent cells were obtained from Agilent Technologies (USA).
The uracil-independent and ampicillin-resistant shuttle vector pJRoC30
was from the California Institute of Technology (CALTECH, USA). The
protease-deficient S. cerevisiae strain
BJ5465 was obtained from the LGCPromochem (Barcelona, Spain). The P. pastoris strain BG11 was purchased to Atum (USA).
The plasmid used (pBSY5Z) was provided by Bisy (Austria). Restriction
endonucleases EcoRI, XbaI, PmeI, BamHI, and XhoI;
the DNA Ligation Kit; the Antarctic phosphatase; and the PNGase F
were purchased from New England Biolabs (USA). iProof High-Fidelity
DNA Polymerase was purchased from Bio-Rad (USA). Oligonucleotide primers
and UPO genes were acquired from Integrated DNA Technologies (USA).
The NucleoSpin plasmid kit and NucleoSpin Gel and PCR Clean-up kit
were purchased from Macherey Nagel (Germany). ABTS was purchased to
Panreac AppliChem (Germany), and DMP and NBD were purchased to TCI
Europe (Switzerland). H₂O₂, styrene, trans β methyl styrene, cis β
methyl styrene, 1,2,3,4-tetrahydronaphtalene, and ethylbenzene were
purchased from Merck Life Science (USA). All chemicals and medium
components were of the highest purity available.

Benzene, toluene, ethylbenzene, potassium sulfate were all from E. Merck (Germany). Xylene isomers purchased from FLUKA (Switzerland). Acetone and THF were supplied from E. Merck. Copper (II) nitrate tri-hydrate, cerium (III) nitrate hexa-hydrate, zirconyl chloride octa-hydrate, PVC powder was purchased from Sigma and is in analytical grade. Copper wire (0.35 mm o.d.) was supplied locally and used after cleaning. Nitrogen and hydrogen gases (99.999%purity) were from Air Products (UK). All the solutions were prepared with doubly distilled water.

Meta-xylene, para-xylene, ortho-xylene, and ethylbenzene chemicals were purchased from Merck company with the purity of minimum 99%. These chemicals are used as received without further purification. Iso-octane was purveyed from Samchun company with the minimum purity of 98%. This material was used as the solvent. Para-diethylbenzene (PDEB) was supplied from Iranian Petrochemical Complex. In the case of the adsorbent, industrial type of Ba-faujasite exchanged namely SPX-3000 with the average particle size of 0.47 mm were utilized.

Wheat and buckwheat flour was purchased from a local supermarket in Dalian, Liaoning, China. Standard chemicals: analytical grade (pentanal, hexanal, heptanal, octanal, nonanal, decanal, 1‐penten‐3‐ol, pentanol, hexanol, heptanol, octanol, toluene, ethyl benzene, p‐xylene, styrene, 2‐pentyl‐furan, and C4–C20 n‐alkanes) were purchased from Sigma‐Aldrich. Internal standards (tetracosane, p‐chloro‐L‐phenylalanine, 5R‐cholestan‐3β‐ol, Phenyl‐β‐D‐glucopyranoside) were purchased from Aladdin.

Reactions were carried out in 50 ml Radleys glass reactors stirred using a Radleys starfish multi reaction system. The reaction mixtures contained 15 U ml_RM⁻¹ PaDa-I, 0.1 mg ml_RM⁻¹ of the metal catalyst (0.5 wt% Au–0.5 wt% Pd/TiO₂) in 100 mM potassium phosphate buffer (pH 6) and 10 mM of substrate (cyclohexane, ethylbenzene, Isophorone, propylbenzene, tetralin and styrene all sourced from Sigma–Aldrich >97%). The catalysts were weighed out directly into the glass vessels and then the buffer was added. Immediately before starting the reactions, enzyme and substrate were added. The system was then pressurized to 2 bar of 80% H₂ 20% air from a high-pressure gas reservoir. The reactions were stirred with a magnetic stirrer bar at 250 and ambient temperature (19.5–20.5 °C). Product formation was monitored by extracting with 2 × 5 ml ethyl acetate containing 2 mM 1-decanol as internal standard and subjecting samples to GC analysis with the concentrations determined through comparisons to a known calibration factor (Supplementary Method 2). Repeat tests—indicated in Fig. 2D—show that measured reaction products agree ± 0.2 mM as determined by GC (2% of the initial reactant concentration of 10 mM) on duplicate experiments.

Benzene, toluene, ethylbenzene, dichloromethane, and ethanol were procured from Sigma-Aldrich, St. Louis, MI, USA. The synthesis of the receptor sensing CME was previously reported by us [27 (link)]. All aqueous solutions of the aromatic compounds were prepared as saturated solutions using deionized (DI) water with a resistivity of 18.6 MΩ·cm. The pH of the deionized water was 6.82. The analyte solutions had a pH of 6.85. The Au-coated QTFs were purchased from Forien Inc., Edmonton, AB, Canada (https://www.fourien.com/). They were coated with Au using a vacuum evaporation method; the thickness of the gold coating was around 100 nm. The resonance frequency of the QTFs was 32.768 kHz; the spring constant was ~20 kN/m; the load capacitance was 12.5 pF. A 10⁻⁶ M solution of the CME was used to functionalize the Au-coated QTFs.

Benzene, toluene, ethylbenzene, dichloromethane, and ethanol were procured from Sigma-Aldrich, St. Louis, MI, USA. The synthesis of the receptor sensing CME was previously reported by us [27 (link)]. All aqueous solutions of the aromatic compounds were prepared as saturated solutions using deionized (DI) water with a resistivity of 18.6 MΩ·cm. The pH of the deionized water was 6.82. The analyte solutions had a pH of 6.85. The Au-coated QTFs were purchased from Forien Inc., Edmonton, AB, Canada (https://www.fourien.com/). They were coated with Au using a vacuum evaporation method; the thickness of the gold coating was around 100 nm. The resonance frequency of the QTFs was 32.768 kHz; the spring constant was ~20 kN/m; the load capacitance was 12.5 pF. A 10⁻⁶ M solution of the CME was used to functionalize the Au-coated QTFs.