Ampicillin sodium salt

Safflower oil was purchased from Gefro (Germany) and had a fatty acid composition of 77.2% linoleic acid, 13.3% oleic acid, 2.4% stearic acid, 6.7% palmitic acid, and 0.4% of other fatty acids [11 (link)]. 13S-HPODE, 12-oxo-9(Z)-dodecenoic acid, and 12-oxo-10(E)-dodecenoic acid standards were from Larodan (Sweden). Hexanal, hexanol, and 12-hydroxydodecanoic acid standards, Glycine max LOX-1, and P. fluorescens Amano lipase were obtained from Sigma-Aldrich (USA). linoleic acid was purchased from Thermo Fisher Scientific (USA), and δ-aminolevulinic acid (ALA), Triton X-100, isopropyl β-d-1-thiogalactopyranoside (IPTG), as well as kanamycin sulfate and ampicillin sodium salt were obtained from Carl Roth (Germany). All other solvents and chemicals were supplied by Carl Roth (Germany), Sigma-Aldrich (USA), or Thermo Fisher Scientific (USA). For PCR, Phusion Hot Start DNA polymerase with 5 × Phusion High-fidelity buffer and dNTPs were obtained from Thermo Fisher Scientific (USA). PageRuler™ Prestained Protein ladder, restriction enzymes, and the monoclonal AP-conjugated Anti-His (C-term) antibody (AB_2556555) were purchased from Thermo Fisher Scientific (USA) as well. Alkaline phosphatase was obtained from New England Biolabs GmbH (Germany).

The 12-aminododecanoic acid standard was purchased from Alfa Aesar (Haverhill, MA, USA), while the 12-oxo-9(Z)-dodecenoic acid and 12-oxo-10(E)-dodecenoic acid standards were purchased from Larodan (Solna, Sweden). Pyridoxal-5-phosphate monohydrate (PLP) was obtained from Acros Organics, Thermo Fisher Scientific (Waltham, MA, USA). Hexanal, hexylamine, soybean LOX-1, and L-lactate dehydrogenase (LDH) were supplied from Sigma Aldrich (St. Louis, MO, USA). Linoleic acid was obtained from Thermo Fisher Scientific (Waltham, MA, USA). β-Nicotine amide adenine dinucleotide disodium salt (NADH), isopropyl β-d-1-thiogalactopyranoside (IPTG), l-alanine, imidazole, ampicillin sodium salt, and kanamycin sulfate were purchased from Carl Roth (Karlsruhe, Germany). 13S-Hydroperoxy-9(Z),11(E)-octadecadienoic acid (13S-HPODE) was prepared from Linoleic acid by a peroxidation reaction with LOX-1 from Glycine max as described previously (Gala Marti et al. 2021 (link)). All other chemicals were from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher Scientific (Waltham, MA, USA) or Carl Roth (Karlsruhe, Germany).

PCR products were either directly cloned or first separated on a 1% low melting gel (Nusieve GTG Agarose, Cambrex, Rockland, USA) excised and cloned with Topo TA Cloning kit (Life Technologies Europe B.V. (Invitrogen), Zug, Switzerland) according to manufacturer’s instruction. After transformation into One Shot Top 10 chemically competent E. coli (Life Technologies Europe B.V. (Invitrogen), Zug, Switzerland) bacteria were grown on a LB-amp-X-Gal plate overnight at 37°C. Four to ten colonies were picked and grown overnight at 37°C in LB medium supplemented with 100 μg ampicillin/ml (Ampicillin-sodium salt, Carl Roth GMBH, Karlsruhe, Germany).
Plasmid DNA purification was performed with the QuickLyse Miniprep Kit (Qiagen AG, Hombrechtikon, Switzerland). The sequencing was done by Microsynth (Balgach, Switzerland). Sequences were edited and analyzed with the following software: DNASTAR software (version 5), clone manager, ClustalX 2.0.3. Phylogenetic trees were constructed with Mega (version 5.1) software
[30 (link)]. Sequence data were submitted to GenBank and were assigned the accession numbers [GenBank: KF990530 to GenBank: KF990540].