Sp 700

Data of optical rotations, UV, and ECD spectra were obtained using Jasco P2000, JASCO V-650, and Jasco J-815 spectrophotometers (Jasco Corporation, Tokyo, Japan), respectively. IR spectra were measured with a Nicolet 5700 spectrometer (Thermo Nicolet Corporation, Madison, SD, USA). GC was performed with an Agilent 7890A instrument (Agilent Technologies, Waldbronn, Germany). The 1D- and 2D-NMR spectra were obtained at 500 or 600 MHz for ¹H and at 125 or 150 MHz for ¹³C, using Bruker 600 and 500 MHz spectrometers (Bruker Corporation, Karlsruhe, Germany). HRESIMS data were acquired with an Agilent 1100 series LC/MSD ion trap mass spectrometer (Agilent Technologies, Waldbronn, Germany). Column chromatography was performed using macroporous resin (Diaion HP-20 and SP-700, from Mitsubishi Chemical Corp., Tokyo, Japan and Sephadex LH-20 columns Pharmacia Fine Chemicals, Uppsala, Sweden. Preparative HPLC was carried out with a Shimadzu LC-6AD instrument with an SPD-20A detector Shimadzu Corp., Tokyo, Japan, using a YMC-Pack ODS-A column (250 mm × 20 mm, 5 μm; YMC Corp., Kyoto, Japan). HPLC-DAD analysis was performed using an Agilent 1200 series system (Agilent Technologies, Waldbronn, Germany) with an Apollo C18 column (250 mm × 4.6 mm, 5 μm; Alltech Corp., Lexington, KY, USA).

Vanillin extracted from vanilla plant pods by NADES was purified by adsorption chromatography using SP700 (Mitsubishi Chemical) non-polar resin by the method previously described (30 (link)). To select the best solvent for purification water and ethanol were individually tested and extraction yield was calculated. Different pH values were also tested to select the optimal pH for vanillin purification. Vanillin from lignin was also purified using optimal solvent and pH by SP700 resin. Microbial vanillin was synthesized by lignin degrading bacteria Bacillus ligniniphilus L1 using the method described in our previous study (31 (link)). Microbial vanillin was extracted from the fermentation broth by previously described method (32 (link)) and purified by employing three molecular weight cut-offs of 50, 5, and 1 kDa tubular ceramic membranes in a membrane fractionation process followed by chromatography using SP700 resin. Purified samples were analyzed by HPLC method described in section “2.4 Experimental design for optimization of vanillin extraction rate” and also identified by GC-MS analysis according the method reported in our previous study (31 (link)).