Orbitrap fusion lumos tribrid mass spectrometer lc ms

Samples comprising 1 mL of the reaction mixture were obtained, and the supernatants were used for the analysis of the relevant metabolites including organic acids and residual glucose, using an Agilent 1260 series high performance liquid chromatography (HPLC) system (Agilent, USA), equipped with a refractive index detector and a Bio-Rad Aminex HPX-87H ion exclusion column (300 mm × 7.8 mm, Aminex, USA), which was kept at 35 °C and eluted with 5 mM H₂SO₄ at a flow rate of 0.5 mL/min; the injection volume was 20 μL [20 (link)]. The cathode was poised at − 650 mV in all the experiments. The curve of current versus time (i–t curve) was used to detect current changes during the entire process of BES reactions. The SDS-PAGE was run using the commercially purchased SurePage™ Gels (GenScript, Nanjing). The protein mass spectrometry was performed using the OrbiTrap Fusion LUMOS Tribrid Mass Spectrometer (LC–MS) (Thermo Fisher, USA) and the methods could be referred to references [21 (link), 22 (link)].

The significance of differences between mean values of control and test samples was assessed using Student’s t-test in the open-source software suite “R” (http://cran.r-project.org/). Differences with p < 0.05 were regarded as obvious, p < 0.01 as significant, and p < 0.001 as very significant. The SDS-PAGE was run using the commercially purchased SurePage™ Gels (GenScript, Nanjing). The protein mass spectrometry was performed using an OrbiTrap Fusion LUMOS Tribrid Mass Spectrometer (LC–MS) (Thermo Fisher, USA) according to published methods [3 (link), 30 (link)].

The significance of differences between mean values of control and test samples was compared using Student's t test in the open‐source software suite “R” (http://cran.r-project.org/). Differences with p < .05 were regarded as obvious, p < .01 as significant, and p < .001 as very significant. The SDS‐PAGE was run using the commercially purchased SurePageTM Gels (GenScript). The protein mass spectrometry was performed using the Orbitrap Fusion Lumos Tribrid Mass Spectrometer (LC‐MS) (Thermo Fisher), and the methods could be referred to references (Espadas, Borras, Chiva, & Sabido, 2017; Li, Zhou, Xiao, Li, & Tian, 2018).