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4 protocols using co 965 column oven

1

Analytical Techniques for Spectroscopic Characterization

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Optical rotations were measured on a JASCO DIP-1000 (JASCO Co., Ltd., Tokyo, Japan). Electrospray ionization-mass spectrometry spectra were obtained using a JEOL JMS-600W spectrometer (JEOL Ltd., Tokyo, Japan) (Supplementary Fig S3). NMR spectra were obtained using a Bruker AVANCE-400 (400.13 MHz for 1H) spectrometer (Bruker Bio Spin K. K., Kanagawa, Japan). The multiplicity of the signals was abbreviated as follows: s = singlet, d = doublet, dd = doublet of doublets, dt = doublet of triplets, t = triplet, q = quartet, m = multiplet, and br = broad. Column chromatography was conducted using sephadex LH-20 resin (GE healthcare Japan Co., Ltd., Tokyo, Japan). Thin-layer chromatography (silica gel, Merck Ltd., Japan) was conducted using van Urk reagent (Ehmann, 1977 ). Preparative high-performance liquid chromatography (HPLC) was conducted using an LC-20AT prominence pump (flow rate 2 mL min-1, Shimadzu Corp., Kyoto, Japan) and an Inertsil ODS-P column (10 × 250 mm, GL science Inc., Tokyo, Japan) in a CO-965 column oven (30 °C, JASCO Co., Ltd., Tokyo, Japan,) and monitored with an SPD-20A UV detector (Shimadzu Corp., Kyoto, Japan). A BioShaker PERSONAL-11 (TAITEC Co., Ltd., Saitama, Japan) was used for the incubation of A. take.
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Size-Exclusion Chromatography of Polymers

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SEC was performed using a high-performance liquid chromatography system consisting of an AS-2057i Plus autosampler (Jasco, Tokyo, Japan), DG-2080-53 degasser (Jasco), PU-2080i Plus pump (Jasco), CO-965 column oven (Jasco), RI-2031 Plus refractive index detector (Jasco), and a combination of TSKgel G4000PWXL and G2500PWXL columns (300 mm × 7.8 mm internal diameter) (Tosoh, Tokyo, Japan). The sample solutions (10 mg/mL, 50 μL) were injected into the system and eluted with 100 mM NaNO3 solution at a flow rate of 1 mL/min at 40 °C. The relative number-average molecular weight (Mn,SEC) and molecular weight distribution (Mw/Mn) were calculated from a calibration curve of standard poly(ethylene glycol) (PEG; Agilent Technologies, Wilmington, DE, USA).
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SEC Characterization of Polymers

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SEC was performed out on an HLC-8120 system (Tosoh, Tokyo, Japan) equipped with a combination of TSKgel AW-4000 and AW-2500 columns (150 mm × 6 mm ID) (Tosoh), eluted with dimethylsulfoxide (DMSO) containing 10 mM LiBr at a flow rate of 0.15 mL/min at 65 °C. The polydispersity index (Mw/Mn) was calculated from a calibration curve of standard PEGs (Agilent Technologies, Wilmington, DE, USA). For SEC with fluorescence detection, the measurements were performed on a Gulliver system (Jasco, Tokyo, Japan) consisting of a DG-2080–53 degasser (Jasco), a PU-980 pump (Jasco), an AU-950 autosampler (Jasco), a CO-965 column oven (Jasco), an FP-920 fluorescence detector (excitation wavelength: 488 nm, emission wavelength: 515 nm) (Jasco), an RI-2031 Plus refractive index detector (Jasco), and a combination of TSKgel α-4000 and TSKgel α-2500 columns (300 mm × 7.8 mm ID) (Tosoh). The solutions (50 μL) were injected into the SEC system, and the system was eluted with a mixture of water and DMSO (volume ratio 50:50) at a flow rate of 0.3 mL/min at 40 °C. 1H NMR spectra were recorded on a Bruker Avance III 500 MHz spectrometer (Bruker BioSpin, Rheinstetten, Germany). FTIR spectra were recorded on a Spectrum 100 FTIR spectrometer (Perkin Elmer, Wellesley, MA, USA). The sample powder was mixed with KBr and pellets were prepared for FTIR measurements.
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Characterization of Macromolecular Structures

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1H nuclear magnetic resonance (NMR) spectra were recorded in D2O at 25 °C using a Bruker Avance III 400 MHz spectrometer (Bruker BioSpin, Rheinstetten, Germany). Fourier transform infrared (FT-IR) spectra were recorded on a Spectrum 100 FT-IR spectrometer (Perkin Elmer, Wellesley, MA, USA). The samples were ground to powder form, which was blended with KBr to prepare pellets for measurements. Aqueous-phase size exclusion chromatography (SEC) measurements were performed using a high-performance liquid chromatography setup consisting of an AU-950 autosampler (Jasco, Tokyo, Japan), PU-980 pump (Jasco, Tokyo, Japan), DG-2080-53 degasser (Jasco, Tokyo, Japan), CO-965 column oven (Jasco, Tokyo, Japan), RI-2031 Plus refractive index detector (Jasco, Tokyo, Japan), and a combination of TSKgel G4000PWXL and G2500PWXL columns (300 mm × 7.8 mm ID; Tosoh, Tokyo, Japan). The sample solutions (10 mg mL−1, 50 μm) were injected into the system and eluted with a 100 × 10−3 M NaNO3 solution at a flow rate of 0.4 mL min−1 at 40 °C [30 (link)].
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