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使用酶法,化学(甲基化)和物理(质谱,核磁共振)技术,植物墙Heteroxylans测序
JoVE 杂志
化学
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JoVE 杂志 化学
Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical (Methylation) and Physical (Mass Spectrometry, Nuclear Magnetic Resonance) Techniques
DOI:

11:49 min

March 24, 2016

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Chapters

  • 00:05Title
  • 00:53Labeling of the Reducing End (RE) Sugar Residue of W-sol AXs with 2-aminobenzamide (2AB)
  • 02:11Generation of Xylo-oligosaccharides from 2 AB Labeled W-sol AXs
  • 03:34Matrix-assisted Laser Desorption Ionization-time of Flight Mass Spectrometry (MALDI-TOF MS)
  • 04:48Electrospray Ionization Quadrupole Time-of-flight Mass Spectrometry (ESI-QTOF MS)
  • 06:20ESI-MSn
  • 07:30H1 Nuclear Magnetic Resonance (NMR) Spectroscopy
  • 08:45Results: Reducing End (RE) and Internal Region Glycosyl Sequence(s) of Heteroxylans
  • 10:34Conclusion

Summary

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This protocol describes the specific techniques used for the structural characterization of reducing end (RE) and internal region glycosyl sequence(s) of heteroxylans by tagging the RE with 2 aminobenzamide prior to enzymatic (endoxylanase) hydrolysis and then analysis of the resultant oligosaccharides using mass spectrometry (MS) and nuclear magnetic resonance (NMR).

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