1260 infinity ri detector

Biomass growth was controlled by optical density (OD₆₀₀) measurement at 600 nm (BioSpectrophotometer, Eppendorf, Hamburg, Germany). The concentration of glycerol and methanol in the cell-free supernatant during the fermentation was determined by HPLC (Agilent Technologies 1220 Infinity LC System with Agilent Technologies 1260 Infinity RI detector, Agilent Technologies, Santa Clara, CA, USA) using a WATREX Polymer IEX H form column and guard column. A flow rate of 0.8 mL/min of 9 mM sulfuric acid at 45 °C was used [38 (link)]. Sinigrin concentration was determined by HPLC (Agilent 1260 Infinity LC System with Quaternary Pump and UV detector, Agilent Technologies, Santa Clara, CA, USA) using a Phenomenex Gemini^® NX 5μm C18 110Å (150 × 4.6 mm) column according to Tsao et al. [39 (link)]. The nano-liquid chromatography and mass spectrometry analysis (LC-MS-MS) of the tryptic digest of myrosinase (myr-Δ19) was modified from [23 (link)]. The detailed method description can be found in Supplementary Materials. Protein concentration in cell-free supernatants was determined using the Bradford method [40 (link)]. Bradford reagent was purchased from Sigma-Aldrich (St. Louis, MO, USA).

Glucose and organic acids were analysed by high-performance liquid chromatography (HPLC) on an Agilent 1290 LC II system, equipped with an Agilent 1290 Infinity Binary Pump, Agilent 1290 Infinity Autosampler, Agilent 1290 Infinity diode array detector operated at 210 nm, and an Agilent 1260 Infinity RI detector operated at 45 °C. Either an Aminex HPX-87H (Bio-Rad) or an Rezex ROA-Organic Acid H + (Phenomenex) column was used with a mobile phase of 0.008 mM H₂SO₄. The HPLC was operated at 0.8 mL/min and 60 °C. Propionic acid (50 mM) was used as internal standard.
Ethyl acetate and ethanol in liquid samples were measured by an Agilent 7890B gas chromatograph equipped with a flame ionisation detector (GC-FID) and an Agilent 7693 autosampler. Samples were analysed by injecting 0.5 μL of liquid sample onto a Nukol™ column (30 m × 0.53 mm, 1.0 μm coating, Supelco). The column temperature was maintained at 50 °C for 2 min and increased to 200 °C at a rate of 50 °C/min. The split ratio was 10. 1-Butanol (2 mM) was used as the internal standard.

Biomass growth was controlled by optical density (OD₆₀₀) measurement at 600 nm (BioSpectrophotometer, Eppendorf, Germany). A calibration curve was then used to calculate the dry cell weight (DCW): DCW (g/L) = 0.2001 × OD₆₀₀ + 0.1075 [21 (link)].
The concentration of glycerol and methanol during fermentation was determined in the cell-free supernatant by HPLC (Agilent Technologies 1220 Infinity LC System with Agilent Technologies 1260 Infinity RI detector, Agilent Technologies, Santa Clara, CA, USA) using WATREX Polymer IEX H form 8 μm (250 × 8 mm column) and WATREX Polymer IEX H form 8 μm (40 × 8 mm guard column). A flow rate of 0.8 mL/min of 9 mM sulfuric acid at 45 °C was used.
The Sinigrin concentration was determined by HPLC (Agilent 1260 Infinity LC System with Quaternary Pump and UV detector, Agilent Technologies, Santa Clara, CA, USA) according to Tsao et al. [42 (link)]. A Phenomenex Gemini^® NX 5μm C18 110Å (150 × 4.6 mm) was used at 30 °C. Sinigrin was purchased from Sigma-Aldrich (Saint-Louis, MO, USA).
The protein concentration in cell-free supernatants was determined by the Bradford method [43 (link)]. The Bradford reagent was purchased from Sigma-Aldrich (Saint-Louis, MO, USA).