Cell growth was determined by measuring the OD600 using a Beckman Coulter DU370 spectrophotometer. Methanol concentrations were measured using an Agilent 1290 Infinity System (Santa Clara, CA, USA) equipped with an Aminex HPX-87H column. l -Lysine was determined using SBA-40E immobilized enzyme biosensor (Shandong, China) [21 (link)]. NADH and NADPH were assayed by means of the NAD/NADH quantitation Kit and the NADP/NADPH quantitation Kit (Sigma-Aldrich).
LC–MS/MS analyses were performed using an UHPLC system (1290, Agilent Technologies) equipped with a UPLC BEH Amide column (1.7 μm, 2.1*100 mm, Waters) coupled to Triple TOF 6600 (Q-TOF, AB Sciex). The mobile phase consisted of 25 mM NH4OAc and 25 mM NH4OH in water (pH = 9.75) (A), and acetonitrile (B) was carried with elution gradient as follows: 0 min-95% B, 0.5 min-95% B, 7 min-65% B, 8 min-40% B, 9 min-40% B, 9.1 min-95%, and B, 12 min-95% B, which was delivered at 0.5 mL/min. The injection volume was 1 μL. The Triple TOF mass spectrometer was used to assess its ability to acquire MS/MS spectra on an information-dependent basis (IDA) during a LC/MS experiment. ESI source conditions were set as follows: Ion source gas 1 as 60, Ion source gas 2 as 60, Curtain gas as 35, source temperature 550 °C, Ion Spray Voltage Floating (ISVF) 5500 V or − 4500 V in positive or negative modes, respectively.
LC–MS/MS analyses were performed using an UHPLC system (1290, Agilent Technologies) equipped with a UPLC BEH Amide column (1.7 μm, 2.1*100 mm, Waters) coupled to Triple TOF 6600 (Q-TOF, AB Sciex). The mobile phase consisted of 25 mM NH4OAc and 25 mM NH4OH in water (pH = 9.75) (A), and acetonitrile (B) was carried with elution gradient as follows: 0 min-95% B, 0.5 min-95% B, 7 min-65% B, 8 min-40% B, 9 min-40% B, 9.1 min-95%, and B, 12 min-95% B, which was delivered at 0.5 mL/min. The injection volume was 1 μL. The Triple TOF mass spectrometer was used to assess its ability to acquire MS/MS spectra on an information-dependent basis (IDA) during a LC/MS experiment. ESI source conditions were set as follows: Ion source gas 1 as 60, Ion source gas 2 as 60, Curtain gas as 35, source temperature 550 °C, Ion Spray Voltage Floating (ISVF) 5500 V or − 4500 V in positive or negative modes, respectively.
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